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Colposcopy is a test performed to detect precancerous lesions of cervical cancer. Since cervical cancer progresses slowly, finding and treating precancerous lesions helps prevent cervical cancer. In particular, it is clinically important to detect high-grade squamous intraepithelial lesions (HSIL) that require surgical treatment among precancerous lesions of cervix. There have been several studies using convolutional neural network (CNN) for classifying colposcopic images. However, no studies have been reported on using the segmentation technique to detect HSIL. In present study, we aimed to examine whether the accuracy of a CNN model in detecting HSIL from colposcopic images can be improved when segmentation information for acetowhite epithelium is added. Without segmentation information, ResNet-18, 50, and 101 achieved classification accuracies of 70.2%, 66.2%, and 69.3%, respectively. The experts classified the same test set with accuracies of 74.6% and 73.0%. After adding segmentation information of acetowhite epithelium to the original images, the classification accuracies of ResNet-18, 50, and 101 improved to 74.8%, 76.3%, and 74.8%, respectively. We demonstrated that the HSIL detection accuracy improved by adding segmentation information to the CNN model, and the improvement in accuracy was consistent across different ResNets. [ABSTRACT FROM AUTHOR]

In a previous study, we showed that several octahydroindene derivatives are potent protease activated receptor 1 (PAR1) antagonists. In the current study, we prepared a series of trans‐fused 5,5‐difulorooctahydroindenes which do not have a C5 stereogenic center, and evaluated their biological activities and metabolic stabilities. Compound 19 in this series, containing a spirooxazolidinone moiety at C2, showed excellent efficacy in both PAR1 binding (IC50 = 70 nM) and human platelet rich plasma (PRP) aggregation (IC50 = 0.19 μM), along with good metabolic stability (R50 = 345.8, 337.2, and 43.4 min in human, rat, and monkey liver microsomes, each), which is comparable to that of vorapaxar. Four stereoisomers of 19 were prepared and evaluated. (1S,2R,3aR,7aR)‐5,5‐Difluoro‐1‐[(E)‐2‐[5‐(3‐fluorophenyl)pyridin‐2‐yl]vinyl]octahydrospiro(indene‐2,5′‐oxazolidin)‐2′‐one (31) was found to be the most active (IC50 = 21 nM) stereoisomer as PAR1 antagonist. Compound 31 exhibited good in vivo oral PK profiles and significant ex vivo antithrombotic efficacy in cynomolgus monkeys upon oral administration. When the plasma concentration of 31 was maintained above 200 ng/mL, platelet aggregation induced by haTRAP was completely inhibited. [ABSTRACT FROM AUTHOR]

Red seaweed, known as Rhodophyta, has a long history of use in traditional Asian medicine, including Traditional Chinese Medicine and Ayurveda. It is believed to have cooling and detoxification properties. Red seaweed species, such as Gracilaria, have been used in traditional remedies to address various conditions, such as inflammation, thyroid disorders, and digestive issues. Obesity is a risk factor of hepatic steatosis, a hallmark of non-alcoholic fatty liver disease (NAFLD) that affects nearly 25% of the worldwide population. Gracilaria chorda (GC) contains bioactive peptides that may be applicable in the prevention of metabolic syndrome diseases. This study investigated the effects of GC subcritical water extract at 210 °C (GCSW210) on preventing liver injury and lipid and glucose dysregulation in an oleic acid (OA)-induced hepatic steatosis cell model (HepG2) and high-fat diet (HFD)-induced obese animal model (C57BL/6J mice). Human hepatoma HepG2 cells were exposed to 0.1 mM OA for 24 h to induce hepatic steatosis and C57BL/6J mice were fed a HFD for 13 weeks. For lipid accumulation, triglyceride (TG) content was measured in both models, along with free fatty acid (FFA), plasma glucose, and insulin levels in HFD-fed mice. Protein expression of master regulators of adipogenesis and lipogenesis, as well as cholesterol and mitochondrial biosynthesis, was studied via western blotting in hepatic steatosis-induced in vitro and in vivo models. In addition, protein expression of the insulin signaling cascade in skeletal muscle tissues of HFD-fed mice was studied. GCSW210 significantly decreased lipid accumulation in HepG2 cells exposed to OA and suppressed the expression of lipogenic factors, such as sterol regulatory element-binding protein (SREBP)-1c and fatty acid synthase. In addition, GCSW210 abrogated transcription factors related to cholesterol biosynthesis, such as SREBP-2 and low-density lipoprotein receptor. Similarly, FFA, TG, serum glutamic acid, aspartate transaminase, alanine transferase, plasma glucose, and insulin levels were also significantly reduced in GCSW210-treated HFD-fed mice, which were comparable to the positive control mice treated with Garcinia cambogia extract. Additionally, GCSW210 enhanced the phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase in the hepatic tissues of HFD-fed mice. Moreover, GCSW210 treatment improved insulin signal transduction by reducing insulin receptor substrate 1 Ser307 phosphorylation and elevated phosphatidylinositol 3-kinase/protein kinase B and glucose transporter type 4 protein expression in muscle tissue. 5-Hdroxymethylfufural (5-HMF) was confirmed to be active substances isolated from GCSW210 through LC-PDA and LC-MS. GCSW210 significantly regulated glucose metabolism, alleviated insulin resistance (IR) induced by high fatty acid synthesis and lipid accumulation, and elevated de novo lipogenesis by activating AMPK phosphorylation in both the liver and muscle tissues of HFD-fed mice. GCSW210 may be a potential functional food for preventing HFD-induced metabolic diseases, such as IR, NAFLD, and type 2 diabetes mellitus. [Display omitted] • GCSW210 alleviated lipid accumulation, fatty acid synthesis, and weight gain. • GCSW210 combat DNL by activating p-AMPK in liver and muscle tissues of HFD-mice. • GCSW210 modulates HFD-induced insulin resistance. • Thus, it improves metabolic diseases such as IR, NAFLD, and T2DM. • The major compound of GCSW210 is 5-HMF. [ABSTRACT FROM AUTHOR]

Several different flexible end effectors have been developed to solve the problem of approaching the lesion in a minimally invasive surgery. In this paper, we developed a wire-driven end effector device to treat frozen shoulder. Since the device is for capsular release surgery, it has a suitable bend radius for the surgery. It is a cylindrical cannula that can fit various surgical tools and can be sterilized after use. The end effector is made of an elastic material called PAI (polyamide-imide) with its outer diameter and total length being 4 and 19 mm. It is controlled by wires that are connected to a motor. Through quantitative evaluation, we confirmed that the end effector can bend up to 90° in an upward or downward direction. Through qualitative evaluation, we confirmed that the device can easier access all regions of the glenoid in a shoulder model than conventional electrocautery. An experiment on a cadaver followed, which allowed us to discuss the real life performance, operation, and areas of improvement of the device with surgeons. From the experiments, we confirmed that our target region, the IGHL (inferior glenohumeral ligament), is within the reach of our device. The surgeon also evaluated that the control of the device caused no inconvenience. [ABSTRACT FROM AUTHOR]

Cisplatin (CDDP) and doxorubicin (DOX) are chemotherapeutic drugs that trigger apoptosis by inducing DNA-damage. A previous study using breast cancer cells demonstrated the negative feedback modulation of the epidermal growth factor receptor (EGFR) and receptor tyrosine-protein kinase erbB-2 (ErbB2) via extracellular signal-regulated kinase (ERK)-mediated phosphorylation of conserved Thr-669 and Thr-677 residues, respectively, in the juxtamembrane domain. In addition, CDDP has been identified to cause negative feedback inhibition of activated EGFR in lung cancer cells. In the present study, the role of phosphorylation in the feedback control of the ErbB2/ErbB3 heterodimer in human breast and gastric cancer cells was investigated. Phosphorylation of ErbB2 at Thr-677 was induced by CDDP and DOX, which in turn reduced tyrosine autophosphorylation of ErbB2 and ErbB3. Treatment with trametinib, a mitogen-activated protein kinase inhibitor that blocks ERK-mediated Thr-677 phosphorylation, and substitution of Thr-677 to alanine, blocked the feedback inhibition of ErbB2 and ErbB3. In addition, these agents caused the degradation of ErbB proteins through the activation of p38 mitogen-activated protein kinase (p38) and ERK. These results demonstrate that chemotherapeutic agents trigger ERK- and p38-mediated post-translational downregulation of ErbB receptors. [ABSTRACT FROM AUTHOR]

Deficiencies in erythrocyte metabolic enzymes are associated with hereditary hemolytic anemia. Here, we report the development of a novel multiplex enzyme assay for six major enzymes, namely glucose-6-phosphate dehydrogenase, pyruvate kinase, pyrimidine 5′-nucleotidase, hexokinase, triosephosphate isomerase, and adenosine deaminase, deficiencies in which are implicated in erythrocyte enzymopathies. To overcome the drawbacks of traditional spectrophotometric enzyme assays, the present assay was based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC–MS/MS). The products of the six enzymes were directly measured by using ion pairing UPLC–MS/MS, and the precision, linearity, ion suppression, optimal sample amounts, and incubation times were evaluated. Eighty-three normal individuals and 13 patients with suspected enzymopathy were analyzed. The UPLC running time was within 5 min. No ion suppression was observed at the retention time for the products or internal standards. We selected an optimal dilution factor and incubation time for each enzyme system. The intra- and inter-assay imprecision values (CVs) were 2.5–12.1% and 2.9–14.3%, respectively. The linearity of each system was good, with R 2 values > 0.97. Patient samples showed consistently lower enzyme activities than those from normal individuals. The present ion paring UPLC–MS/MS assay enables facile and reproducible multiplex evaluation of the activity of enzymes implicated in enzymopathy-associated hemolytic anemia. [ABSTRACT FROM AUTHOR]

Maternal embryonic leucine zipper kinase ( MELK) has been implicated in various cellular processes and is highly upregulated in diverse cancers, then it is thought to be a promising anticancer target. 3-Anilino-1 H-pyrazolo[3,4- b]pyridine scaffold was identified as a novel scaffold for MELK kinase inhibitors in high throughput screening ( HTS). From exploration for structure-activity relationship ( SAR) studies mainly by the modification of the 3 (C3), and 5-positions (C5), 4-(4-methylpiperazin-1-yl)- N-{5-[1-(piperidin-4-yl)-1 H-pyrazol-4-yl]-1 H-pyrazolo[3,4- b]pyridin-3-yl}benzamide ( 21a) was found to exhibit good activity (IC50 = 0.15 μM) on MELK as a lead in early state. [ABSTRACT FROM AUTHOR]

Sodium dichloroisocyanurate-96% (NaDCC) is commonly used to treat drinking water, industrial water, and wastewater. However, exposure to NaDCC by inhalation can have toxic pulmonary effects in humans. In the present study, we evaluated the potential toxicity of NaDCC following a 90-day inhalation toxicity study in Sprague-Dawley/Crl:CD (SD) rats. The animals were exposed to 0.4, 2.0, or 10.0 mg/m3 NaDCC for 90 days. In addition, male and female rats from the 10.0 mg/m3 group were set up as the recovery group for 14 days. The bronchoalveolar lavage fluid showed a concentration-dependent increase in the total cell count, with a significant increase in neutrophils in both the sexes in the 10.0 mg/m3 group compared to the negative control group. In the 10.0 mg/m3 group, lung organ weight was significantly increased among the female rats. Histopathological examination showed eosinophilic droplets in the olfactory/respiratory epithelium, mucous cell hyperplasia, atrophy/degeneration of the tracheal branches, and wall thickening of the alveolar ducts in the nasal cavity of both sexes in the 10.0 mg/m3 group. The adverse effects of NaDCC exposure were observed to decrease during the 14-day recovery period in both sexes. Based on pathological observations, the "no observed adverse effect concentration (NOAEC)" of inhaled NaDCC was 2.0 mg/m3 for both sexes. These results are expected to provide a scientific basis for inhalation toxicity data of NaDCC. [Display omitted] • The total cell number in BALF increased in a concentration-dependent manner. • The histological changes showed in nasal cavity and bronchi of 10.0 mg/m3 group. • In the alveolar duct, the adverse effect of wall thickening by NaDCC was observed. • The adverse effect was observed to decrease during the recovery period. • The no-observed adverse effect concentration of NaDCC was 2.0 mg/m3 for both sexes. [ABSTRACT FROM AUTHOR]

Objectives Development of acute kidney injury (AKI) after cardiopulmonary bypass (CPB) is relatively common and associated with increased mortality. Recently, plasma neutrophil gelatinase-associated lipocalin (NGAL) was used for the prediction of AKI. We evaluated the clinical usefulness of plasma NGAL. Design and methods One hundred twelve adult patients undergoing cardiovascular surgery with CPB were included. Blood samples were obtained at baseline, at intensive care unit (ICU) admission, and 24 h after ICU admission. The development of AKI, which is defined as an increase in serum creatinine by more than 50% within 3 postoperative days, was monitored. NGAL levels were analyzed by a Biosite Triage meter (Alere Medical, USA). Diagnostic performance of NGAL was analyzed using the area under the receiver operating characteristic curve. Results In AKI patients (n = 13), plasma NGAL levels at ICU admission were significantly higher than those at baseline [177 (122–402) vs. 121 (74–158) ng/mL, median (interquartile range), p = 0.028], whereas serum creatinine showed no significant change. The predictive value of NGAL at ICU admission was 0.812 [95% confidence interval (CI), 0.68 to 0.95] with a cut-off value of 168.5 ng/mL (sensitivity, 61.5%; specificity, 88.9%). After the exclusion of 35 patients with preoperative decreased renal function, the predictive value was increased to 0.911 (95% CI, 0.82 to 1.00). Conclusions This study showed that plasma NGAL may serve as a useful biomarker for the early detection of AKI in adult patients following CPB. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: There is sparse data on apoB dyslipoproteinemia in Asian population. The purpose of this study was to assess apoB dyslipoproteinemia and to compare the LDL-C, non-HDL and apoB for risk assessment with percentile equivalent cut off in Korean population. Methods: With 1193 Korean adult subjects, the prevalence and characteristics of different types of dyslipoproteinemias were analyzed in each age and gender group. The percentile values of direct LDL-C, calculated LDL-C, non HDL-C, HDL-C, apoAI, apoB and apoB/apoAI ratio were estimated. Results: The prevalences of normoapoB–hyperTG, hyperapoB–normoTG and hyperapoB–hyperTG dyslipoproteinemia were 6.9, 8.9 and 10.9% in men and 3.7, 6.4 and 2.8% in women. The 40th percentile of direct LDL-C, calculated LDL-C, non-HDL-C and apo B were 108, 104.2, 126 and 85mg/dl, respectively. The individual above optimal cut off was significantly underestimated with LDL-C than with non-HDL and apoB, in groups with adverse risk factors. Conclusions: This study firstly shows the prevalence of various types of dyslipoproteinemias in Asian population. The percentile values of Korean population were similar to those of NHANES. Integration of lipid markers is needed for making clinical decisions and further research involving various populations and methodologies should be performed. [Copyright &y& Elsevier]

Abstract: Piperazinyl derivatives of 1-(arylsulfonyl)-2,3-dihydro-1H-quinolin-4-ones have been identified with high binding affinities for 5-HT6 receptor. In particular, 2-methyl-5-(N-methyl-piperazin-1-yl)-1-(naphthalene-2-sulfonyl)-2,3-dihydro-1H-quinolin-4-one (8g) exhibits high binding affinity toward 5-HT6 (IC50 =8nM) receptor with good selectivity over other serotonin and dopamine receptors. [ABSTRACT FROM AUTHOR]

Abstract: (Piperazin-1-yl-phenyl)-arylsulfonamides were synthesized and identified to show high affinities for both 5-HT2C and 5-HT6 receptors. Among them, naphthalene-2-sulfonic acid isopropyl-[3-(4-methyl-piperazin-1-yl)-phenyl]-amide (6b) exhibits the highest affinity towards both 5-HT2C (IC50 =4nM) and 5-HT6 receptors (IC50 =3nM) with good selectivity over other serotonin (5-HT1A, 5-HT2A, and 5-HT7) and dopamine (D2–D4) receptor subtypes. In 5-HT2C and 5-HT6 receptor functional assays, this compound showed considerable antagonistic activity for both receptors. [ABSTRACT FROM AUTHOR]

Abstract: A novel series of 1H-indole-3-carboxylic acid pyridine-3-ylamides were synthesized and identified to show high affinity and selectivity for 5-HT2C receptor. Among them, 1H-indole-3-carboxylic acid[6-(2-chloro-pyridin-3-yloxy)-pyridin-3-yl]-amide (15k) exhibits the highest affinity (IC50 =0.5nM) with an excellent selectivity (>2000 times) over other serotonin (5-HT1A, 5-HT2A, and 5-HT6) and dopamine (D2–D4) receptors. [Copyright &y& Elsevier]

[Display omitted] • Pycnogenol® lessened lung weight gain caused by polyhexamethylene guanidine (PHMG). • Reduced inflammation caused by lung fibrosis was also observed. • The expression of fibrosis-related factors was reduced by Pycnogenol®. • Pycnogenol® improved histopathological changes in the lungs of PHMG-treated mice. • Pycnogenol® may hinder the progression of pulmonary fibrosis. Pulmonary fibrosis (PF) is a respiratory disease that causes serious respiratory problems. The effects of French marine pine bark extract (Pycnogenol®), with antioxidant and anti-inflammatory properties, were investigated on lung fibrosis in polyhexamethylene guanidine (PHMG)-treated mice. Mice were separated into four groups (n = 6): vehicle control (VC, saline 50 μl); PHMG (1.1 mg/kg); PHMG + Pycnogenol® (0.3 mg/kg/day); and PHMG + Pycnogenol® (1 mg/kg/day). PF was induced via intratracheal instillation of PHMG. Treatment with PHMG decreased body weight and increased lung weight, both of which were improved by treatment with PHMG + Pycnogenol® (1 mg/kg). Enzyme-linked immunosorbent assay, western blotting, and PCR revealed that Pycnogenol® attenuated PHMG-induced increase in inflammatory cytokines and fibrosis-related factors in a dose-dependent manner. Finally, histopathological analysis revealed reduced inflammation/fibrosis in the PHMG + Pycnogenol® (1 mg/kg) group. Collectively, the results indicate that Pycnogenol® can be used to treat PF as it hinders fibrosis progression by inhibiting inflammatory responses in the lungs of PHMG-treated mice. [ABSTRACT FROM AUTHOR]

More realistic effects on glucose metabolic dysfunction can be evaluated by applying organochlorine (OCP) mixtures than individual OCPs. We formulated an equal ratio mixture of five OCPs (chlordane, heptachlor, p,p′-dichlorodiphenyltrichloroethane, β-hexachlorocyclohexane, and hexachlorobenzene) and treated L6 myotubes with this OCP mixture to investigate effects on glucose uptake and the underlying mechanism. Exposure to the OCP mixture reduced 2-NBDG staining, representing glucose uptake, and stimulated the excessive production of reactive oxygen species (ROS). Reduced 2-NBDG uptake and ROS overproduction were compensated by insulin treatment. The expression of proteins such as IRβ, PI3K, and AKT was downregulated, indicating that ROS overproduction contributed to the inhibition of insulin-dependent glucose uptake. Reduction in mitochondria quantity and decreased expression levels of PGC-1α, PDH, and GLUT4 proteins were observed, suggesting that mitochondrial dysfunction played a causative role in the disruption of glucose uptake. The inhibition of glucose uptake and ROS overproduction caused by the OCP mixture were also found in zebrafish as an in vivo model. We demonstrated that exposure to the OCP mixture, even at the lowest concentration, perturbed glucose uptake, which was associated with mitochondrial dysfunction, suggesting that an OCP mixture could be a potential environmental factor in type 2 diabetes-related effects on skeletal muscles. [Display omitted] • The OCP mixture inhibits glucose uptake in L6 myotubes. • The OCP mixture stimulates the excessive production of ROS in L6 myotubes. • Expression of proteins in insulin signaling and mitochondrial function is reduced. • Glucose uptake and ROS overproduction is also observed in zebrafish. • A low concentration of OCP mixture can be the major environmental factor for type 2 diabetes-related effects. [ABSTRACT FROM AUTHOR]

Abstract: A concise and versatile method for the preparation of 3-aminoindoles from 2-(disubstituted amino)-benzonitriles is described, and in situ functionalizations were illustrated. [Copyright &y& Elsevier]

While aminoacyl-tRNA synthetase-interacting multifunctional protein 2 (AIMP2) is a tumor suppressor, its exon 2-depleted splice variant (AIMP2-DX2 or shortly DX2) is highly expressed in human lung cancer, and the ratio of DX2 to AIMP2 increases according to the progression of lung cancer. In this study, pyrimethamine inhibited the level of DX2 (IC50 = 0.73 µM) in A549 cells expressing nanoluciferase-tagged DX2. In a panel of 5 lung cancer cell lines with various DX2 levels, pyrimethamine most potently suppressed the growth of H460 cells, which express high levels of DX2 (GI50 = 0.01 µM). An immunoblot assay in H460 cells showed that pyrimethamine decreased the DX2 level dose-dependently but did not affect the AIMP2 level. Further experiments confirmed that pyrimethamine resulted in ubiquitination-mediated DX2 degradation. In an in vivo mouse xenograft assay using H460 cells, intraperitoneal administration of pyrimethamine significantly reduced the tumor size and weight, comparable with the effects of taxol, without affecting body weight. Analysis of tumor tissue showed a considerably high concentration of pyrimethamine with a decreased levels of DX2. These results suggest that pyrimethamine, currently used as anti-parasite drug, could be repurposed to treat lung cancer patients expressing high level of DX2. [ABSTRACT FROM AUTHOR]

We selected five substances among the organochlorine pesticides (OCPs), chlordane, heptachlor, p,p'-DDT, β-HCH, and hexachlorobenzene, and investigated whether low-concentration exposure to the OCP compounds affected glucose metabolism. The exposure of L6 myotubes to the OCP compounds (1 or 5 μM) for 24 and 48 h significantly inhibited glucose uptake with an excessive production of intracellular reactive oxygen species (ROS) and peroxynitrite anions (ONOO−) compared to control cells. In contrast, the production of nitric oxide was highly reduced by exposure to the OCP compounds. The protein expression of glucose transporter 4 (GLUT4) in the L6 myotubes was significantly suppressed by exposure to the OCP compounds. In addition, exposure to the OCP compounds for 1 h in RIN-m5F pancreatic beta cells remarkably suppressed insulin secretion but the ability to secrete insulin recovered to control levels after 24 h exposure to the OCP compounds. The abundant ROS generated by 1 h exposure to OCP compounds was inversely related to insulin secretion in RIN-m5F pancreatic beta cells. Therefore, these results suggest that low-concenration exposure of skeletal muscle and pancreatic beta cells to OCP compounds may affect insulin secretion and insulin-dependent glucose uptake through extreme oxidative stress and inactivation of the glucose transport protein. • Low-dose exposure of skeletal muscle and pancreatic beta cells to OCP compounds inhibits insulin secretion and insulin-dependent glucose uptake. • OCP compounds induced excessive production of ROS and ONOO− in pancreatic beta cells and skeletal muscle cells. • OCP compounds are a major factor in the development of insulin resistance and type 2 diabetes. [ABSTRACT FROM AUTHOR]

New therapies for treating drug-resistant pneumococcal infections are urgently needed. The novel scaffold 6-hydroxy-4-oxo-1,2-dihydro-4 H -quinoline was shown to have similar efficacies against all three different serotypes of S. pneumoniae , ATCC 49617™ (19F), ATCC BAA-1663™ (15B), and ATCC 700904™ (19A), in a resazurin-based high-throughput screen using the Korea Chemical Bank library. Further studies to identify a new lead with this scaffold, including tricyclic pyrrolo[3,2,1- ij ]quinolone and pyrido[3,2,1- ij ]quinolone derivatives, led to the identification of 6d , 7d and 12a. Compound 6d (IC 50 = 0.92, 0.75, and 0.77 µM), 7d (IC 50 = 0.57, 0.66, and 0.38 µM) and 12a (IC 50 = 0.27, 1.03, and 0.62 µM) showed submicromolar IC 50 values against 19F, 15B, and 19A, respectively, and thus serve as a starting point for further optimization. While some of compounds in this series exhibited acceptable pharmacokinetic profiles in preliminary in vivo rat experiments, the most active compound 12a showed poor solubility and high plasma protein binding. Our current research efforts are focused on optimizing compounds to improve physicochemical properties as well as potency. [ABSTRACT FROM AUTHOR]

Trans -activator (Tat)-mediated human immunodeficiency virus type 1 (HIV-1) transcription is essential for the replication of HIV-1 and is considered a potent therapeutic target for HIV-1 inhibition. In this study, the Library of Pharmacologically Active Compounds (LOPAC1280) was screened using our dual-reporter screening system for repositioning as Tat-inhibitory compounds. Consequently, two compounds were found to be potent, with low cytotoxicity. Of these two compounds, Roscovitine (CYC202) is already known to be a Tat inhibitor, while gemcitabine has been newly identified as an inhibitor of Tat-mediated transcription linked to viral production and replication. In an additional screening using the ribonucleoside analogues of gemcitabine, two analogues (2′-C-methylcytidine and 3-deazauridine) showed a specific Tat-inhibitory effect linked to their anti-HIV-1 activity. Interestingly, these compounds did not affect Tat protein directly, while the mechanism underlying their inhibition of Tat-mediated transcription was linked to pyrimidine biosynthesis, rather than to alteration of the dNTP pool, influenced by the inhibition of ribonucleotide reductase. Taken together, the proposed functional screening system is a useful tool for the identification of inhibitors of Tat-mediated HIV-1 transcription from among a large number of compounds, and the inhibitory effect of HIV-1 transcription by gemcitabine and its analogues may suggest a strategy for developing a new class of therapeutic anti-HIV drugs. • Several Tat-inhibitory compounds were identified from a library of pharmacological agents. • Of these, gemcitabine and its analogues exhibited a novel Tat inhibitory effect. • Their Tat-inhibitory mechanism was linked to the inhibition of pyrimidine biosynthesis. • Their novel mode of action could provide a strategy for developing new anti-HIV drugs. [ABSTRACT FROM AUTHOR]

Background: The present study aimed to evaluate the long-term oncological and obstetric outcomes following the loop electrosurgical excision procedure (LEEP) in patients with cervical intraepithelial neoplasia (CIN) and investigate the risk factors for recurrence and preterm birth. Methods: This retrospective cohort study included patients who underwent LEEP for CIN 2–3 between 2011 and 2019. Demographic information, histopathological findings, postoperative cytology, and human papillomavirus (HPV) status were collected and analyzed. The Cox proportional hazards model and Kaplan-Meier curves with the log-rank test were used for risk factor analysis. Results: A total of 385 patients treated with the LEEP were analyzed. Treatment failure, including recurrence or residual disease following surgery, was observed in 13.5% of the patients. Positive surgical margins and postoperative HPV detection were independent risk factors for CIN1 + recurrence or residual disease (HR 1.948 [95%CI 1.020–3.720], p = 0.043, and HR 6.848 [95%CI 3.652–12.840], p-value < 0.001, respectively). Thirty-one patients subsequently delivered after LEEP, and the duration between LEEP and delivery was significantly associated with preterm-related complications, such as a short cervix, preterm labor, and preterm premature rupture of the membrane (p = 0.009). However, only a history of preterm birth was associated with preterm delivery. Conclusions: Positive HPV status after LEEP and margin status were identified as independent risk factors for treatment failure in patients with CIN who underwent LEEP. However, combining these two factors did not improve the prediction accuracy for recurrence. [ABSTRACT FROM AUTHOR]

Human immunodeficiency virus-1 (HIV-1) transactivator (Tat)-mediated transcription is essential for HIV-1 replication. It is determined by the interaction between Tat and transactivation response (TAR) RNA, a highly conserved process representing a prominent therapeutic target against HIV-1 replication. However, owing to the limitations of current high-throughput screening (HTS) assays, no drug that disrupts the Tat-TAR RNA interaction has been uncovered yet. We designed a homogenous (mix-and-read) time-resolved fluorescence resonance energy transfer (TR-FRET) assay using europium cryptate as a fluorescence donor. It was optimized by evaluating different probing systems for Tat-derived peptides or TAR RNA. The specificity of the optimal assay was validated by mutants of the Tat-derived peptides and TAR RNA fragment, individually and by competitive inhibition with known TAR RNA-binding peptides. The assay generated a constant Tat-TAR RNA interaction signal, discriminating the compounds that disrupted the interaction. Combined with a functional assay, the TR-FRET assay identified two small molecules (460-G06 and 463-H08) capable of inhibiting Tat activity and HIV-1 infection from a large-scale compound library. The simplicity, ease of operation, and rapidity of our assay render it suitable for HTS to identify Tat-TAR RNA interaction inhibitors. The identified compounds may also act as potent molecular scaffolds for developing a new HIV-1 drug class. [ABSTRACT FROM AUTHOR]

Background Although polymyxin has been used as a last-resort antibiotic against resistant bacteria, its use is restricted due to nephrotoxicity and neurotoxicity. While the present antibiotic resistance issue compels clinicians to reconsider polymyxin use in severe illness cases, polymyxin-resistant microorganisms exert an effect. Objectives To address the issue of antibiotic resistance, the cycle of developing new antibiotics to counteract emerging resistance must be discontinued. Here we tried to develop novel therapies that do not rely on direct antimicrobial activity and thus do not promote antibiotic resistance. Methods By a high-throughout screening system based on bacterial respiration, chemical compounds accelerating the antimicrobial effects of polymyxin B were screened. In vitro and in vivo tests were performed to validate adjuvanticity. In addition, membrane depolarization and total transcriptome analysis were used to determine molecular mechanisms. Results PA108, a newly discovered chemical compound, was used to eradicate polymyxin-resistant A. baumannii and three other species in the presence of polymyxin B at concentrations less than the MIC. Since this molecule lacks self-bactericidal action, we hypothesized that PA108 acts as an antibiotic adjuvant, enhancing the antimicrobial activity of polymyxin B against resistant bacteria. At working concentrations, no toxicity was observed in cell lines or mice, although co-treatment with PA108 and polymyxin B increased survival of infected mouse and decreased bacterial loads in organs. Conclusions Boosting antibiotic efficiency through the use of antibiotic adjuvants holds significant promise for tackling the rise in bacterial antibiotic resistance. [ABSTRACT FROM AUTHOR]

[Display omitted] • The series of N -arylindazole-3-carboxamides were designed and synthesized. • Total 20 novel compounds were evaluated against SARS-CoV-2. • The most promising compound 4a showed an EC 50 of 0.69 μM with no cytotoxicity (CC 50 > 25 μM). • Compound 4a exhibited satisfactory in vitro PK profiles as well. N -Arylindazole-3-carboxamide derivatives synthesized from an anti-MERS-CoV hit compound showed potent inhibitory activities against SARS-CoV-2. Among them, 5-chloro- N -(3,5-dichlorophenyl)-1 H -indazole-3-carboxamide (4a) exhibited a potent inhibitory effect (EC 50 = 0.69 µM), low cytotoxicity, and satisfactory in vitro PK profiles. Thus, N -arylindazole-3-carboxamide 4a provides a novel template for future development of anti-coronavirus agents. [ABSTRACT FROM AUTHOR]

The heterocyclic indole structure has been shown to be one of the most promising scaffolds, offering various medicinal advantages from its wide range of biological activity. Nonetheless, the significance of 3-oxindole has been less known. In this study, a series of novel 3-oxindole-2-carboxylates were synthesized and their antiviral activity against human immunodeficiency virus-1 (HIV-1) infection was evaluated. Among these, methyl (E)-2-(3-chloroallyl)-4,6-dimethyl-one (6f) exhibited the most potent inhibitory effect on HIV-1 infection, with a half-maximal inhibitory concentration (IC50) of 0.4578 μM but without severe cytotoxicity (selectivity index (SI) = 111.37). The inhibitory effect of these compounds on HIV-1 infection was concordant with their inhibitory effect on the viral replication cycle. Mode-of-action studies have shown that these prominent derivatives specifically inhibited the Tat-mediated viral transcription on the HIV-1 LTR promoter instead of reverse transcription or integration. Overall, our findings indicate that 3-oxindole derivatives could be useful as a potent scaffold for the development of a new class of anti-HIV-1 agents. [ABSTRACT FROM AUTHOR]

Lgr5 has been identified as a marker of the stem/progenitor cells in the murine ovary and oviduct by lineage tracing. However, little is known regarding LGR5 expression or its functional significance in human ovary tissues. Here, using RNA in situ hybridization and/or immunohistochemistry, we thoroughly investigated LGR5 expression in normal human ovaries, fallopian tubes and various ovarian tumors. We discovered that LGR5 expression is negligible in the human ovary surface epithelium, whereas ovarian stromal cells normally express low levels of LGR5. Remarkably, fallopian tube epithelium, inclusion cysts and serous cystadenomas with a Müllerian phenotype expressed high levels of LGR5, and LGR5 expression was restricted to PAX8+/FOXJ1− secretory cells of the tubal epithelium. Strong stromal LGR5 expression without epithelial LGR5 expression was consistently observed in the path from serous cystadenoma to serous borderline tumor to low grade serous carcinoma (LGSC). Unlike LGSC, high grade serous carcinoma (HGSC), clear cell carcinoma, endometrioid carcinomas displayed various epithelial-stromal LGR5 expression. Notably, high levels of LGR5 expression were observed in serous tubal intraepithelial carcinoma, which slightly declined in invasive HGSC. LGR5 expression was significantly associated with improved progression-free survival in HGSC patients. Moreover, in vitro assays demonstrated that LGR5 expression suppressed tumor proliferation and migratory capabilities. Taken together, these findings indicate a tumor-suppressive role for LGR5 in the progression of HGSC. [ABSTRACT FROM AUTHOR]

We previously reported the potent antiviral effect of the 2-aminoquinazolin-4-(3H)-one 1, which shows significant activity (IC50 = 0.23 μM) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with no cytotoxicity. However, it is necessary to improve the in vivo pharmacokinetics of compound 1 because its area under the curve (AUC) and maximum plasma concentration are low. Here, we designed and synthesized N-substituted quinazolinone derivatives that had good pharmacokinetics and that retained their inhibitory activity against SARS-CoV-2. These compounds were conveniently prepared on a large scale through a one-pot reaction using Dimroth rearrangement as a key step. The synthesized compounds showed potent inhibitory activity, low binding to hERG channels, and good microsomal stability. In vivo pharmacokinetic studies showed that compound 2b had the highest exposure (AUC24h = 41.57 μg∙h/mL) of the synthesized compounds. An in vivo single-dose toxicity evaluation of compound 2b at 250 and 500 mg/kg in rats resulted in no deaths and an approximate lethal dose greater than 500 mg/kg. This study shows that N-acetyl 2-aminoquinazolin-4-(3H)-one 2b is a promising lead compound for developing anti-SARS-CoV-2 agents. [ABSTRACT FROM AUTHOR]

Abstract: A 5,7-dichloro-3-phenyl-3-methyl-quinoline-2,4-dione (11a) has been identified in a random screen as a lead for 5-HT6 antagonist. During the lead optimization process, several analogs were synthesized and their biological activities were investigated. Within this series, several compounds display high binding affinity and selectivity for the 5-HT6 receptor. In particular, 3-(4-hydroxyphenyl)-3-methyl-quinoline-2,4-dione (12f) exhibits high affinity (K i =12.3nM) for 5-HT6 receptor with good selectivity over other serotonin and dopamine (D1–D4) receptor subtypes. In a functional adenylyl cyclase stimulation assay, this compound exhibited considerable antagonistic activity (IC50 =0.61μM). [Copyright &y& Elsevier]

Despite the continuing global crisis caused by coronavirus disease 2019 (COVID‐19), there is still no effective treatment. Therefore, we designed and synthesized a novel series of 2‐benzylaminoquinazolin‐4(3H)‐one derivatives and demonstrated that they are effective against SARS‐CoV‐2. Among the synthesized derivatives, 7‐chloro‐2‐(((4‐chlorophenyl)(phenyl)methyl)amino)quinazolin‐4(3H)‐one (Compound 39) showed highest anti‐SARS‐CoV‐2 activity, with a half‐maximal inhibitory concentration value greater than that of remdesivir (IC50 = 4.2 μM vs. 7.6 μM, respectively), which gained urgent approval from the U.S. Food and Drug Administration. In addition, Compound 39 showed good results in various assays measuring metabolic stability, human ether a‐go‐go, Cytochromes P450 (CYPs) inhibition, and plasma protein binding (PPB), and showed better solubility and pharmacokinetics than our previous work. [ABSTRACT FROM AUTHOR]

We screened a variety of natural products against MERS-CoV as an attempt of developing I anti i -MERS-CoV drugs, and many flavonoids showed I anti i -MERS activities. Total 12 compounds of bavachinin derivatives in four core structures were evaluated to figure out their I anti i -MERS-CoV activity and cell-cytotoxicity by cellular phenotypic screening method as shown in Table. B 2a b was prepared by hydrogenations of external double bonds of B 1a b , which showed the best I anti i -MERS activities regardless of their I O i -substituents. The I anti i -MERS activity of B 6a b was a little decreased compared to B 1a b , however, the I anti i -MERS activities of its derivatives were conserved. [Extracted from the article]

Graphical abstract Highlights • Osthenol was a potent (IC 50 = 0.74 µM) and selective inhibitor for hMAO-A (SI > 81.1). • Osthenol was a reversible and competitive inhibitor (K i = 0.26 µM). • The binding affinity of osthenol for hMAO-A was greater than that for hMAO-B. • The 8-(3,3-dimethylallyl) group of osthenol increased its inhibitory activity against hMAO-A. • Osthenol can be a potential lead compound for design of novel reversible MAO-A inhibitors. Abstract Osthenol (6), a prenylated coumarin isolated from the dried roots of Angelica pubescens , potently and selectively inhibited recombinant human monoamine oxidase-A (hMAO-A) with an IC 50 value of 0.74 µM and showed a high selectivity index (SI > 81.1) for hMAO-A versus hMAO-B. Compound 6 was a reversible competitive hMAO-A inhibitor (K i = 0.26 µM) with a potency greater than toloxatone (IC 50 = 0.93 µM), a marketed drug. Isopsoralen (3) and bakuchicin (1), furanocoumarin derivatives isolated from Psoralea corylifolia L., showed slightly higher IC 50 values (0.88 and 1.78 µM, respectively) for hMAO-A than 6 , but had low SI values (3.1 for both). Other coumarins tested did not effectively inhibit hMAO-A or hMAO-B. A structural comparison suggested that the 8-(3,3-dimethylallyl) group of 6 increased its inhibitory activity against hMAO-A compared with the 6-methoxy group of scopoletin (4). Molecular docking simulations revealed that the binding affinity of 6 for hMAO-A (−8.5 kcal/mol) was greater than that for hMAO-B (−5.6 kcal/mol) and that of 4 for hMAO-A (−7.3 kcal/mol). Docking simulations also implied that 6 interacted with hMAO-A at Phe208 and with hMAO-B at Ile199 by carbon hydrogen bondings. Our findings suggest that osthenol, derived from natural products, is a selective and potent reversible inhibitor of MAO-A, and can be regarded a potential lead compound for the design of novel reversible MAO-A inhibitors. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Three flavanones and two flavones were isolated from the leaves of Prunus padus var. seoulensis. • Rhamnocitrin was a potent hMAO-A inhibitor (IC 50 = 0.051 µM), the lowest natural flavonoid to date. • The binding affinity of 5 for hMAO-A was greater than that for hMAO-B. • Genkwanin (4) strongly inhibited hMAO-A and hMAO-B (IC 50 = 0.14 and 0.35 µM, respectively). • Compounds 5 and 4 can be considered as new potent and reversible hMAO-A and/or hMAO-B inhibitors. Abstract Three flavanones and two flavones were isolated from the leaves of Prunus padus var. seoulensis by the activity-guided screening for new monoamine oxidase (MAO) inhibitors. Among the compounds isolated, rhamnocitrin (5) was found to potently and selectively inhibit human MAO-A (hMAO-A, IC 50 = 0.051 µM) and effectively inhibit hMAO-B (IC 50 = 2.97 µM). The IC 50 value of 5 for hMAO-A was the lowest amongst all natural flavonoids reported to date, and the potency was 20.2 times higher than that of toloxatone (1.03 µM), a marketed drug. In addition, 5 reversibly and competitively inhibited hMAO-A and hMAO-B with K i values of 0.030 and 0.91 µM, respectively. Genkwanin (4) was also observed to strongly inhibit hMAO-A and hMAO-B (IC 50 = 0.14 and 0.35 µM, respectively), and competitively inhibit hMAO-A and hMAO-B (K i = 0.097 and 0.12 µM, respectively). Molecular docking simulation reveals that the binding affinity of 5 with hMAO-A (−18.49 kcal/mol) is higher than that observed with hMAO-B (0.19 kcal/mol). Compound 5 interacts with hMAO-A at four possible residues (Asn181, Gln215, Thr336, and Tyr444), while hMAO-B forms a single hydrogen bond at Glu84. These findings suggest that compound 5 as well as 4 can be considered as novel potent and reversible hMAO-A and/or hMAO-B inhibitors or useful lead compounds for future development of hMAO inhibitors in neurological disorder therapies. [ABSTRACT FROM AUTHOR]

Seaweeds are considered a potential source of antiobesity agents. Because Caulerpa , a seaweed, has been consumed for food in Japan, China, South Korea, and Australia, we hypothesized that Caulerpa okamurae may have antiobesity effects in an animal model of high-fat diet (HFD)–induced obesity in C57BL/6 mice. Herein, we found that the ethanolic extract of C okamurae (COE) significantly inhibited lipid accumulation and reduced the expression of the master regulator of adipogenesis, peroxisome proliferator-activated receptor-γ, sterol regulatory element binding protein-1c, and CCAAT/enhancer-binding protein-α in 3T3-L1 adipocytes. Moreover, COE significantly decreased body weight, fat weight, and liver weight in HFD-fed mice. This effect is comparable to that of positive control Garcinia cambogia extract, which has been approved by the Korean Food and Drug Administration as a weight loss food supplement in South Korea. Similarly, markers of weight gain such as free fatty acids, triglyceride, total cholesterol, glucose, and insulin in the plasma and free fatty acid, triglyceride, total cholesterol, and total lipid in the liver are significantly reduced in COE-treated HFD-fed mice. We found significantly reduced peroxisome proliferator-activated receptor-γ, CCAAT/enhancer-binding protein-α, fatty acid synthase, sterol regulatory element binding protein-1c, cluster of differentiation 36, and acetyl-CoA synthetase in the adipose tissue of COE-treated HFD-fed mice. In conclusion, our results demonstrated that COE is effective in preventing body weight gain and fat accumulation and reduces plasma and hepatic lipid profiles. Together, these findings suggest that C okamurae may be used as a possible treatment option for the management of obesity and associated metabolic disorders. [ABSTRACT FROM AUTHOR]

[Display omitted] The COVID-19 pandemic caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) continues to threaten human health and create socioeconomic problems worldwide. A library of 200,000 small molecules from the Korea Chemical Bank (KCB) were evaluated for their inhibitory activities against SARS-CoV-2 in a phenotypic-based screening assay to discover new therapeutics to combat COVID-19. A primary hit of this screen was the quinolone structure-containing compound 1. Based on the structure of compound 1 and enoxacin, which is a quinolone-based antibiotic previously reported to have weak activity against SARS-CoV-2, we designed and synthesized 2-aminoquinolone acid derivatives. Among them, compound 9b exhibited potent antiviral activity against SARS-CoV-2 (EC 50 = 1.5 µM) without causing toxicity, while having satisfactory in vitro PK profiles. This study shows that 2-aminoquinolone acid 9b provides a promising new template for developing anti-SARS-CoV-2 entry inhibitors. [ABSTRACT FROM AUTHOR]

The role of gold nanoparticles supported on ZnO in photocatalytic activity for dye degradation was investigated. To this end, gold nanoparticles supported on ZnO (Au-ZnO) were prepared using a simple co-precipitation method. The prepared nanocatalyst was characterized by high-resolution transmission electron microscopy, X-ray diffraction, temperature-programmed reduction, X-ray photoelectron spectroscopy, UV-Vis absorption, and photoluminescence. The photocatalytic activity of Au-ZnO was examined by the degradation of methylene blue in aqueous solution using a light source that has more than 95 % (i.e., energy) of emitted photons between 400 and 800 nm. Highly enhanced photocatalytic degradation of methylene blue in air at room temperature was observed from these Au-ZnO nanocatalysts with gold particle size ranging from 2 to 7 nm, with an average size of 3.8 nm. The observed rate constant for MB degradation on Au-ZnO was 0.0118/min compared with 0.0007/min for pure ZnO. Furthermore, the charge transfer pathway for the degradation of methylene blue in Au-ZnO is suggested. [ABSTRACT FROM AUTHOR]

Point-of-care testing (POCT) coagulometers are increasingly being used in the hospital setting. We investigated whether the prothrombin time international normalized ratio (INR) results by CoaguChek XS Plus (Roche Diagnostics GmbH, Mannheim, Germany) can be used reliably without being confirmed with the INR results by STA-R system (Diagnostica Stago S.A.S, Asnières sur Seine, France).~Background~Background~A total of 118 INR measurements by CoaguChek XS Plus and STA-R were compared using Passing/Bablok regression analysis and Bland-Altman plot. Agreement of the INR measurements was further assessed in relation to dosing decision.~Methods~Methods~The correlation of INR measurements between CoaguChek XS Plus and STA-R was excellent (correlation coefficient = 0.964). The mean difference tended to increase as INR results increased and was 0.25 INR in the therapeutic range (2.0-3.0 INR). The overall agreement was fair to good (kappa = 0.679), and 21/118 (17.8%) INR measurements showed a difference in dosing decision.~Results~Results~The positive bias of CoaguChek XS Plus may be obvious even in the therapeutic INR range, and dosing decision based on the CoaguChek XS Plus INR results would be different from that based on the STA-R results. The INR measurements by POCT coagulometers still need to be confirmed with the laboratory INR measurements.~Conclusion~Conclusions [ABSTRACT FROM AUTHOR]

Abstract: Background: Whole blood viscosity (WBV) is the resistance of blood flow in blood vessels. Increased WBV may be a cardiovascular risk factor. The proper screening of WBV can help the early detection of cardiovascular disease. We investigated the performance of a new scanning capillary tube viscometer (SCTV) for the measurement of WBV. Methods: We evaluated the total precision of the SCTV for 20days using three control viscosity materials, and the within-day precision with the whole blood samples of three different individuals. For the linearity evaluation, serial dilutions of a high concentration standard material were used. For the method comparison, the results of the SCTV method were compared to those of Brookfield rotating viscometer on 227 subjects. Results: The SCTV had good within-run and total-run coefficient of variant (CV)s at low-, medium-, and high-concentration samples, at shear rates of 1 and 300s−1. The within-day CVs with the three human blood samples were 6.3%, 3.7% and 3.8% at a shear rate of 1s−1, and 3.2%, 3.0% and 4.1% at a shear rate of 300s−1. The SCTV method showed an excellent linearity in the range of 84.9 to 558.2 milliPoise (mP) and 28.8 to 71.0 mP at shear rates of 1 and 300s−1, respectively. For the comparison study, the SCTV and the rotating viscometer showed comparable results. Conclusions: The SCTV showed a stable analytical performance, and was comparable with the rotational viscometer. This new SCTV method can be used in the clinical laboratory for various needs. [Copyright &y& Elsevier]

Objective: This study aimed to establish reference intervals for lymphocyte subsets including CD4+CD25highFoxP3+ regulatory T-cells (Tregs) in umbilical cord blood. Methods: Umbilical cord blood was obtained after birth from 120 healthy full-term neonates, who were born between November 2010 and November 2011. Lymphocyte subsets including Tregs were analysed using flow cytometer (Beckman Coulter, Fullerton, CA, USA), and the reference intervals were defined using non-parametrical percentile methods according to the Clinical and Laboratory Standard Institute guideline (C28-A3). Results: The reference intervals for lymphocyte subsets were: helper T-cells (CD3+/CD4+), 15.40-70.06%; cytotoxic T-cells (CD3+/CD8+), 9.65-34.28%; B-cells (CD19+), 4.50-29.59%; and natural killer cells (CD3−/CD16+/CD56+), 1.42-28.03%. The reference interval for Tregs was 0.35-9.07%. Conclusions: This study provides the reference intervals for lymphocyte subsets including Tregs in umbilical cord blood from healthy full-term neonates. These results could be used as fundamental data for clinical laboratory tests as well as future researches. [ABSTRACT FROM AUTHOR]

Abstract: Background: Leukocyte reduction filters are widely used for platelet transfusion therapy, and effective leukocyte removal is mandatory for transfusion safety. We evaluated both the performance of leukocyte reduction filters for platelets and the effect of filtration on platelet function. Methods: A total of 100 pooled products (eight platelet concentrates were randomly pooled for each product) were used in this study: 50 were filtered by BioP-plus (Fresenius Kabi AG, Homburg, Germany) and 50 by PXL-8 (Pall Corporation, East Hills, NY, USA). Leukocyte reduction, platelet recovery, and filtration time were evaluated in each leukocyte reduction filter. Platelet aggregation responses to thrombin receptor activation peptide stimulation were compared in pre- and post-filtration products using impedance aggregometry (Multiplate Analyzer, Dynabyte Medical, Munich, Germany). Results: Leukocyte counts were uniformly less than 8.3×105 in all the post-filtration products, except one filtered by the PXL-8. Leukocyte reduction was 99.1% for BioP-plus and 99.7% for PXL-8, and platelet recovery was 84.2% for the BioP-plus and 86.7% for PXL-8. Filtration time of the BioP-plus was significantly shorter than that of PXL-8. Post-filtration platelet aggregation tended to decrease in both filters, showing no difference between them. Conclusions: Both BioP-plus and PXL-8 leukocyte reduction filters for platelets performed well with effective leukocyte reduction and satisfactory platelet recovery. Although platelet function was decreased after filtration procedures, its clinical relevance is uncertain. [Copyright &y& Elsevier]

Abstract: Objectives: The performance of recently developed vitamin D total assays (ADVIA Centaur and Elecsys) was compared to that of liquid chromatography–tandem mass spectrometry (LC–MS/MS) and LIASON 25-OH Vitamin D total assays. Design and methods: A total of 157 clinical samples and standard reference material (SRM) 972 were analyzed. Results: The correlations of LC–MS/MS with the three immunoassays were acceptable. However, compared to LC–MS/MS, LIAISON and ADVIA Centaur showed negative bias, and Elecsys showed positive bias. There was a lack of agreement among the four methods with only LC–MS/MS results close to the certified values of SRM 972. The prevalence of vitamin D insufficiency (<50nmol/L) was higher with ADVIA Centaur (51.6%) and LIAISON (52.2%) and lower with Elecsys (37.6%), compared with that of LC–MS/MS (44.6%). Conclusion: The new, automated total vitamin D assays show acceptable correlation with LC–MS/MS, and could be used in routine laboratories. However, standardization of vitamin D assays and consideration of assay-specific decision limits should be addressed. [Copyright &y& Elsevier]

The rhizome of Dryopteris crassirhizoma Nakai. (Dryopteridaceae) has been used in traditional medicine in East Asia and has recently been reported to have anticancer, anti-inflammation, and antibacterial activity as well as antiviral activity. Natural phloroglucinols from D. crassirhizoma, dryocrassin ABBA and filixic acid ABA were reported to inhibit influenza virus infection with an inhibitory activity on neuraminidase. In this study, we found that dryocrassin ABBA and filixic acid ABA have an inhibitory activity against the main protease of SARS-CoV-2. Therefore, dryocrassin ABBA and filixic acid ABA exhibited inhibitory activity against SARS-CoV-2 infection in Vero cells dose-dependently using the immunofluorescence-based antiviral assays. Moreover, these compounds inhibited SARS-CoV and MERS-CoV infection, suggesting their broad-spectrum anticoronaviral activity. In addition, a 5-day repeated-dose toxicity study of dryocrassin ABBA and filixic acid ABA suggested that an approximately lethal dose of these compounds in mice was >10 mg/kg. Pharmacokinetic studies of dryocrassin ABBA showed good microsomal stability, low hERG inhibition, and low CYP450 inhibition. In vivo pharmacokinetic properties of dryocrassin ABBA showed a long half-life (5.5–12.6 h) and high plasma exposure (AUC 19.3–65 μg·h/mL). Therefore, dryocrassin ABBA has therapeutic potential against emerging coronavirus infections, including COVID-19. [ABSTRACT FROM AUTHOR]

Cardiotonic steroids are steroid-like natural compounds known to inhibit Na+/K+-ATPase pumps. To develop a broad-spectrum antiviral drug against the emerging coronavirus infection, this study assessed the antiviral properties of these compounds. The activity of seven types of cardiotonic steroids against the MERS-CoV, SARS-CoV, and SARS-CoV-2 coronavirus varieties was analyzed using immunofluorescence antiviral assay in virus-infected cells. Bufalin, cinobufagin, and telocinobufagin showed high anti-MERS-CoV activities (IC50, 0.017~0.027 μM); bufalin showed the most potent anti-SARS-CoV and SARS-CoV-2 activity (IC50, 0.016~0.019 μM); cinobufotalin and resibufogenin showed comparatively low anti-coronavirus activity (IC50, 0.231~1.612 μM). Differentially expressed genes in Calu3 cells treated with cinobufagin, telocinobufagin, or bufalin, which had high antiviral activity during MERS-CoV infection were analyzed using QuantSeq 3′ mRNA-Seq analysis and data showed similar gene expression patterns. Furthermore, the intraperitoneal administration of 10 mg/kg/day bufalin, cinobufagin, or digitoxin induced 100% death after 1, 2, and 4 days in 5-day repeated dose toxicity studies and it indicated that bufalin had the strongest toxicity. Pharmacokinetic studies suggested that telocinobufagin, which had high anti-coronavirus activity and low toxicity, had better microsomal stability, lower CYP inhibition, and better oral bioavailability than cinobufagin. Therefore, telocinobufagin might be the most promising cardiotonic steroid as a therapeutic for emerging coronavirus infections, including COVID-19. [ABSTRACT FROM AUTHOR]

Simple Summary: Despite the importance of vaccination against Coronavirus disease 2019 (COVID-19) in cancer patients, general vaccine uptake rates among cancer patients are known to be low. Here, we tried to investigate the attitude and acceptance rates for COVID-19 vaccine in cancer patients and identify predictive factors for vaccination that could be modified to promote vaccine uptake rates. Between February and April 2021, a total of 1001 cancer patients from five institutions participated in a paper-based survey, consisting of 58 items over six domains. Among the respondents, 61.8% were willing to receive the COVID-19 vaccine. Along with the previously reported predictors for COVID-19 vaccination, including male gender, older age, and influenza vaccination history, we distinctively found that patient's disease status and health status (absence of cancer recurrence, time since cancer diagnosis over 5 years, and higher EuroQol Visual Analogue Scale scores) were associated with higher acceptance rates of vaccination. Furthermore, physician's recommendations effectively reduced patient's vaccine hesitancy. Considering the high morbidity and mortality of Coronavirus disease 2019 (COVID-19) in patients with malignancy, they are regarded as a priority for COVID-19 vaccination. However, general vaccine uptake rates among cancer patients are known to be lower than in their healthy counterparts. Thus, we aimed to investigate the attitude and acceptance rates for the COVID-19 vaccine in cancer patients and identify predictive factors for vaccination that could be modified to increase vaccine uptake rates, via a paper-based survey (58 items over six domains). A total of 1001 cancer patients participated in this nationwide, multicenter survey between February and April 2021. We observed that 61.8% of respondents were willing to receive the COVID-19 vaccine. Positive predictive factors found to be independently associated with vaccination were male gender, older age, obesity, previous influenza vaccination history, absence of cancer recurrence, time since cancer diagnosis over 5 years, and higher EuroQol Visual Analogue Scale scores. Along with the well-known factors that are positively correlated with vaccination, here, we report that patients' disease status and current health status were also associated with their acceptance of the COVID-19 vaccination. Moreover, 91.2% of cancer patients were willing to be vaccinated if their attending physicians recommend it, indicating that almost 30% could change their decision upon physicians' recommendation. Unlike other factors, which are unmodifiable, physicians' recommendation is the single modifiable factor that could change patients' behavior. In conclusion, we firstly report that Korean cancer patients' acceptance rate of the COVID-19 vaccination was 61.8% and associated with disease status and current health status. Physicians should play a major role in aiding cancer patients' decision-making concerning COVID-19 vaccines. [ABSTRACT FROM AUTHOR]

[Display omitted] Despite the rising threat of fatal coronaviruses, there are no general proven effective antivirals to treat them. 2-Aminoquinazolin-4(3 H)-one derivatives were newly designed, synthesized, and investigated to show the inhibitory effects on SARS-CoV-2 and MERS-CoV. Among the synthesized derivatives, 7-chloro-2-((3,5-dichlorophenyl)amino)quinazolin-4(3 H)-one (9g) and 2-((3,5-dichlorophenyl)amino)-5-hydroxyquinazolin-4 (3 H)-one (11e) showed the most potent anti-SARS-CoV-2 activities (IC 50 < 0.25 μM) and anti-MERS-CoV activities (IC 50 < 1.1 μM) with no cytotoxicity (CC 50 > 25 μM). In addition, both compounds showed acceptable results in metabolic stabilities, hERG binding affinities, CYP inhibitions, and preliminary PK studies. [ABSTRACT FROM AUTHOR]

Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) continues to spread worldwide, with 25 million confirmed cases and 800 thousand deaths. Effective treatments to target SARS-CoV-2 are urgently needed. In the present study, we have identified a class of cyclic sulfonamide derivatives as novel SARS-CoV-2 inhibitors. Compound 13c of the synthesized compounds exhibited robust inhibitory activity (IC 50 = 0.88 μM) against SARS-CoV-2 without cytotoxicity (CC 50 > 25 μM), with a selectivity index (SI) of 30.7. In addition, compound 13c exhibited high oral bioavailability (77%) and metabolic stability with good safety profiles in hERG and cytotoxicity studies. The present study identified that cyclic sulfonamide derivatives are a promising new template for the development of anti-SARS-CoV-2 agents. [ABSTRACT FROM AUTHOR]

Streptococcus pneumoniae is one of Gram-positive pathogen that causes invasive pneumococcal disease. Nowadays, many S. pneumoniae strains are resistant to commonly used antibiotics such as β-lactams and macrolides. 3-Acyl-2-phenylamino-1,4-dihydroquinolin-4-one (APDQ) derivatives are known as novel chemicals having anti-pneumococcal activity against S. pneumoniae. The underlying mechanism of the anti-pneumococcal activity of this inhibitor remains unknown. Therefore, we tried to find the anti-pneumococcal mechanism of APDQ230122, one of the APDQ derivatives active against S. pneumoniae. We performed transcriptomic analysis (RNA-Seq) and proteomic analysis (LC–MS/MS analysis) to get differentially expressed genes (DEG) and differentially expressed proteins (DEP) of S. pneumoniae 521 treated with sub-inhibitory concentrations of APDQ230122 and elucidated the comprehensive expression changes of genes and proteins using multi-omics analysis. As a result, genes or proteins of peptidoglycan biosynthesis and DNA replication were significantly down-regulated. Electron microscopy analysis revealed that the structure of peptidoglycan was damaged by APDQ230122 in a chemical concentration-dependent manner. Therefore, we suggest peptidoglycan biosynthesis is a major target of APDQ230122. Multi-omics analysis can provide us useful information to elucidate anti-pneumococcal activity of APDQ230122. [ABSTRACT FROM AUTHOR]

New therapies for treating coronaviruses are urgently needed. A series of 4-anilino-6-aminoquinazoline derivatives were synthesized and evaluated to show high anti-MERS-CoV activities. N4 -(3-Chloro-4-fluorophenyl)- N6 -(3-methoxybenzyl)quinazoline-4,6-diamine (1) has been identified in a random screen as a hit compound for inhibiting MERS-CoV infection. Throughout optimization process, compound 20 was found to exhibit high inhibitory effect (IC 50 = 0.157 μM, SI = 25) with no cytotoxicity and moderate in vivo PK properties. [ABSTRACT FROM AUTHOR]

3-Acyl-2-phenylamino-1,4-dihydroquinolin-4(1 H)-one derivatives were synthesized and evaluated to show high anti-MERS-CoV inhibitory activities. Among them, 6,8-difluoro-3-isobutyryl-2-((2,3,4-trifluorophenyl)amino)quinolin-4(1 H)-one (6u) exhibits high inhibitory effect (IC 50 = 86 nM) and low toxicity (CC 50 > 25 μM). Moreover, it shows good metabolic stability, low hERG binding affinity, no cytotoxicity, and good in vivo PK properties. [ABSTRACT FROM AUTHOR]

Stephania tetrandra and other related species of Menispermaceae are the major sources of the bis-benzylisoquinoline alkaloids tetrandrine (TET), fangchinoline (FAN), and cepharanthine (CEP). Although the pharmacological properties of these compounds include anticancer and anti-inflammatory activities, the antiviral effects of these compounds against human coronavirus (HCoV) remain unclear. Hence, the aims of the current study were to assess the antiviral activities of TET, FAN, and CEP and to elucidate the underlying mechanisms in HCoV-OC43-infected MRC-5 human lung cells. These compounds significantly inhibited virus-induced cell death at the early stage of virus infection. TET, FAN, and CEP treatment dramatically suppressed the replication of HCoV-OC43 as well as inhibited viral S and N protein expression. The virus-induced host response was reduced by compound treatment as compared with the vehicle control. Taken together, these findings demonstrate that TET, FAN, and CEP are potential natural antiviral agents for the prevention and treatment of HCoV-OC43 infection. [ABSTRACT FROM AUTHOR]