Recent discovery of natural substances with cathepsin L-inhibitory activity for cancer metastasis suppression.

Cathepsin L (CTSL), a cysteine cathepsin protease of the papain superfamily, plays a crucial role in cancer progression and metastasis. Dysregulation of CTSL is frequently observed in tumor malignancies, leading to the degradation of extracellular matrix and facilitating epithelial-mesenchymal transition (EMT), a key process in malignant cancer metastasis. This review mainly provides a comprehensive information about recent findings on natural inhibitors targeting CTSL and their anticancer effects, which have emerged as potent anticancer therapeutic agents or metastasis-suppressive adjuvants. Specifically, inhibitors are categorized into small-molecule and macromolecule inhibitors, with a particular emphasis on cathepsin propeptide-type macromolecules. Additionally, the article explores the molecular mechanisms of CTSL involvement in cancer metastasis, highlighting its regulation at transcriptional, translational, post-translational, and epigenetic levels. This work underscores the importance of understanding natural CTSL inhibitors and provides researchers with practical insights to advance the relevant fields and discover novel CTSL-targeting inhibitors from natural sources.

Clinical and Immunological Predictors of Hemorrhagic Fever with Renal Syndrome Outcome during the Early Phase.

The ability to accurately predict the early progression of hemorrhagic fever with renal syndrome (HFRS) is crucial for reducing morbidity and mortality rates in severely affected patients. However, the utility of biomarkers for predicting clinical outcomes remains elusive in HFRS. The aims of the current study were to analyze the serum levels of immune function-related proteins and identify novel biomarkers that may help ascertain clinical outcomes of HFRS. Enzyme-linked immunosorbent assay, Luminex, and bioanalyzer assays were used to quantitatively detect 15 biomarkers in 49 serum samples of 26 patients with HFRS. High hemoglobin (HGB) and low urine output (UO) levels were identified as potential biomarkers associated with the acute HFRS. The serum soluble urokinase plasminogen activator receptor (suPAR) and C-X-C motif chemokine ligand 10 (CXCL10) values increased in the early phase of diseases. Elevated suPAR, interleukin-10 (IL-10), CXCL10, and decreased transforming growth factor-beta 3 (TGF-ß3) were representative predictors of the disease severity. Upregulation of the HGB showed a significant correlation with high levels of suPAR and CXCL10. Reduced UO positively correlated with increased suPAR, CXCL10, and TGF-ß2, and decreased vascular endothelial growth factor and TGF-ß3. The changing HGB and UO criteria, high suPAR, IL-10, CXCL10, and low TGF-ß3 of HFRS raise significant awareness for physicians regarding prospective biomarkers for monitoring early warning signs of HFRS. This study provides critical insights into the clinical and immunological biomarkers for disease severity and progression in patients with HFRS to identify early predictions of fatal outcomes.

Morphological Features and Cold-Response Gene Expression in Mesophilic Bacillus cereus Group and Psychrotolerant Bacillus cereus Group under Low Temperature.

At low temperatures, psychrotolerant B. cereus group strains exhibit a higher growth rate than mesophilic strains do. However, the different survival responses of the psychrotolerant strain (BCG34) and the mesophilic strain (BCGT) at low temperatures are unclear. We investigated the morphological and genomic features of BCGT and BCG34 to characterize their growth strategies at low temperatures. At low temperatures, morphological changes were observed only in BCGT. These morphological changes included the elongation of rod-shaped cells, whereas the cell shape in BCG34 was unchanged at the low temperature. A transcriptomic analysis revealed that both species exhibited different growth-related traits during low-temperature growth. The BCGT strain induces fatty acid biosynthesis, sulfur assimilation, and methionine and cysteine biosynthesis as a survival mechanism in cold systems. Increases in energy metabolism and fatty acid biosynthesis in the mesophilic B. cereus group strain might explain its ability to grow at low temperatures. Several pathways involved in carbohydrate mechanisms were downregulated to conserve the energy required for growth. Peptidoglycan biosynthesis was upregulated, implying that a change of gene expression in both RNA-Seq and RT-qPCR contributed to sustaining its growth and rod shape at low temperatures. These results improve our understanding of the growth response of the B. cereus group, including psychrotolerant B. cereus group strains, at low temperatures and provide information for improving bacterial inhibition strategies in the food industry.

Novel Paju Apodemus paramyxovirus 1 and 2, harbored by Apodemus agrarius in the Republic of Korea.

Paramyxoviruses harbored by multiple natural reservoirs pose a potential threat to public health. Jeilongvirus has been proposed as a novel paramyxovirus genus found in rodents, bats, and cats. Paramyxovirus RNA was detected in 108/824 (13.1%) Apodemus agrarius captured at 14 trapping sites in the Republic of Korea. We first present two genetically distinct novel paramyxoviruses, Paju Apodemus paramyxovirus 1 (PAPV-1) and 2 (PAPV-2). The disparity between PAPV-1 (19,716 nucleotides) and -2 (17,475 nucleotides) revealed the presence of the SH gene and length of the G gene in the genome organization. The phylogeny of PAPV-1 and -2 belonged to distinct genetic lineages of Jeilongvirus, respectively, even though these viruses were originated from A. agrarius. PAPV-1 infected human epithelial and endothelial cells, facilitating the induction of type I/III interferons, interferon-stimulated genes, and pro-inflammatory cytokines. Therefore, this study provides insights into the molecular epidemiology, genetic diversity, and virus-host interactions of novel rodent-borne paramyxoviruses.

Variables that prolong total operative time for robotic-assisted laparoscopic myomectomy: A 10-year tertiary hospital study in Korea.

STUDY OBJECTIVE: To identify factors that prolong total operative time (TOT) in robotic-assisted laparoscopic myomectomy (RALM). DESIGN: Retrospective cohort study. SETTING: Tertiary university hospital. PATIENTS: Women who underwent RALM between April 2009 and May 2019 conducted by a single high-volume gynecologic surgeon. INTERVENTIONS: Patients' demographic data and intraoperative records were obtained. The association between the perioperative characteristics and TOT was analyzed. MEASUREMENTS AND MAIN RESULTS: A total of 584 cases met the inclusion criteria, with a mean TOT of 231.6 ± 86.7 min. The mean patient age was 36.3 ± 5.5 years, and the patients had a mean of 4.2 ± 4.0 myomas. The dominant myoma had a mean diameter of 7.6 ± 2.6 cm. The mean total weight of the extracted myomas removed was 202.2 ± 152.6 g. From multiple regression analysis, the following perioperative factors were intimately associated with the TOT: ① body mass index, ② the number of myomas, ③ weight of total myomas, ④ location of dominant myoma, ⑤ type of da Vinci robot system, ⑥ endometrial cavity opening during the operation, ⑦ intraoperative blood loss, and ⑧ patient hospitalization period. The number of myoma was most closely related to the TOT, with an R2 value of 0.330. All of the above factors with the exception of the type of robot system and location of dominant myoma were related to the console time. Age, parity, history of previous abdominal surgery, surgical indication, diameter, and FIGO classification were not associated with the TOT. CONCLUSION: With an accurate identification of the perioperative parameters above, we can improve the quality of RALM by counselling, selecting an appropriate patient selection, and preoperative planning.

Efficient myometrial defect closure in a layer by layer fashion after robot-assisted laparoscopic adenomyomectomy: a novel technique.

OBJECTIVE: In this video, we present our novel technique for myometrial defect closure following robot-assisted laparoscopic adenomyomectomy. METHODS: A narrated video demonstration of our technique. Our patient was a 47-year-old single woman with severe dysmenorrhea, who did not respond to medical therapy and wished to preserve her uterus. Surgery was performed after thorough counseling and obtaining informed consent from the patient (Institutional Review Board number: KC17OESI0238; approval date: March 19, 2018). After removal of the adenomyotic tissue during surgical intervention, the myometrial defect was closed in three steps. First, the defect between the anterior and posterior innermost myometrial layers was closed using a 2-0 Stratafix suture, CT-1 (circle taper) needle (Ethicon, Somerville, NJ, USA). Next, the two sides were approximated using a 2-0 PDS® (polydioxanone) Suture (Ethicon, Somerville, NJ, USA) and V-34 (TAPERCUT®) surgical needle (Ethicon, Somerville, NJ, USA). Finally, the serosa was sutured in a baseball fashion using a 2-0 PDS suture, slim half-circle [SH] needle (Ethicon, Somerville, NJ, USA). RESULTS: The patient had no postoperative complications, and her pain was greatly improved. The CA125 level decreased from 434 U/mL to 45.99 U/mL, and the transvaginal ultrasound showed a reduction in posterior myometrial thickness from 5.61 cm to 2.69 cm. CONCLUSION: This technique maintained the integrity of the endometrial cavity, posterior myometrial thickness, and uterine layer alignment. We believe that it is a feasible technique and may be a solution for adenomyosis in patients seeking for fertility preservation.

Lipase-catalyzed synthesis of lauroyl tripeptide-KHA with multi-functionalities: Its surface-active, antibacterial, and antioxidant properties.

Lipase-catalyzed acylation of a hydrophilic tripeptide-KHA (TP-KHA; amino acid sequence Lys-His-Ala) with a lipophilic lauric acid was performed to produce a multi-functional compound, lauroyl tripeptide-KHA (TPL-KHA), with surface, antibacterial, and antioxidant activities. The significant acylation reaction parameters were optimized as follows: organic solvent of 2-methyl-2-butanol, reaction temperature at 55 °C, substrate molar ratio (lauric acid:TP-KHA) of 4.0, and reaction time for 72 h. Structural analyses by LC-ESI-MS and 1H NMR identified that Nε-lauroyl tripeptide-KHA was chemo-selectively synthesized by the acylation reaction under the optimum conditions. TPL-KHA showed the surface activity at the air-water interface with critical micelle concentration (CMC) of 2.71 mM and γCMC of 30.44 mN/m. TPL-KHA exhibited bacteriostatic and bactericidal effects on Gram-positive and Gram-negative foodborne pathogens (minimum inhibitory concentrations: 2.83-4.00 mM, minimum bactericidal concentrations: 3.17-5.83 mM). Moreover, it was demonstrated that TPL-KHA had the ability to scavenge ABTS+ radicals and inhibit the lipid oxidation.

Phc2 controls hematopoietic stem and progenitor cell mobilization from bone marrow by repressing Vcam1 expression.

The timely mobilization of hematopoietic stem and progenitor cells (HSPCs) is essential for maintaining hematopoietic and tissue leukocyte homeostasis. Understanding how HSPCs migrate between bone marrow (BM) and peripheral tissues is of great significance in the clinical setting, where therapeutic strategies for modulating their migration capacity determine the clinical outcome. Here, we identify an epigenetic regulator, Phc2, as a critical modulator of HSPC trafficking. The genetic ablation of Phc2 in mice causes a severe defect in HSPC mobilization through the derepression of Vcam1 in bone marrow stromal cells (BMSCs), ultimately leading to a systemic immunodeficiency. Moreover, the pharmacological inhibition of VCAM-1 in Phc2-deficient mice reverses the symptoms. We further determine that Phc2-dependent Vcam1 repression in BMSCs is mediated by the epigenetic regulation of H3K27me3 and H2AK119ub. Together, our data demonstrate a cell-extrinsic role for Phc2 in controlling the mobilization of HSPCs by finely tuning their bone marrow niche.

In Vitro Bovine Liver Experiment of Cisplatin-Infused and Normal Saline-Infused Radiofrequency Ablation with an Internally Cooled Perfusion Electrode.

PURPOSE: To evaluate the efficacy of cisplatin-infused and normal saline-infused radiofrequency ablation (RFA) with internally cooled perfusion (ICP) electrode. MATERIALS AND METHODS: Using a 200 W generator, thirty ablation zones were created and divided into three groups of 10 each as follows: group A, RFA alone with 16 gauge monopolar internally cooled (IC) electrode; group B, cisplatin-infused RFA with 16 gauge ICP electrode; and group C, normal saline-infused RFA with 16 gauge ICP electrode. Radiofrequency was applied to the explanted bovine liver for 12 min. During RFA, cisplatin and normal saline were injected into tissue at a rate of 0.5 mL/min through the ICP electrode by injection pump. Dimensions of the ablation zone and technical parameters were compared between the three groups. RESULT: In the cisplatin-infused RFA group, the ablation zone size was significantly larger than that of the RFA-alone group but significantly smaller than normal saline-infused RFA group. The width of longitudinal section and volume were 3.39 ± 0.22 cm2 and 26.55 ± 4.62 cm3 in RFA-alone group, 3.88 ± 0.32 cm2 and 36.45 ± 5.46 cm3 in cisplatin-infused RFA group, and 4.52 ± 0.50 cm2 and 49.44 ± 7.55 cm3 in normal saline-infused RFA group, respectively (p < 0.05 between any two groups). The mean impedance in group A, B, and C were 60.0 ± 7.2, 50.3 ± 2.5, and 40.3 ± 4.0 Ω, respectively (p < 0.05 between any two groups). CONCLUSION: Cisplatin-infused RFA with ICP electrode created the larger size of ablation zone than that of monopolar RFA with an IC electrode, but created the smaller size of ablation zone than that of normal saline-infused RFA.

Lipase-catalyzed solvent-free synthesis of erythorbyl laurate in a gas-solid-liquid multiphase system.

Erythorbyl laurate is a potential food additive as a multi-functional emulsifier having antioxidant and antimicrobial activities. In this study, a gas-solid-liquid multiphase system (GSL-MPS) was established to enhance the production yield of erythorbyl laurate in a lipase-catalyzed solvent-free synthesis. The significant reaction variables were optimized as follows: substrate molar ratio of 2:1 (lauric acid:erythorbic acid) and enzyme concentration of 120 mg/mL (840 PLU/mL). Under these conditions, the maximum production yield in GSL-MPS was 13.974 mg/mL, which is 8.60- and 4.26-fold higher than the yields obtained in an organic solvent monophase system (OS-MPS) and a solid-liquid biphase system (SL-BPS), respectively. Moreover, the operational stability of the immobilized lipase was significantly improved in GSL-MPS compared with OS-MPS. These results indicate that GSL-MPS can be an enzymatic reaction system facilitating efficient production of ester compounds as a means of increasing production yields and the reusability of the immobilized lipase.

Molecular cloning and anti-invasive activity of cathepsin L propeptide-like protein from Calotropis procera R. Br. against cancer cells.

Cathepsin L of cancer cells has been shown to play an important role in degradation of extracellular matrix for metastasis. In order to reduce cell invasion, cathepsin L propeptide-like proteins which are classified as the I29 family in the MEROPS peptidase database were characterized from Calotropis procera R. Br., rich in cysteine protease. Of 19 candidates, the cloned and expressed recombinant SnuCalCp03-propeptide (rSnuCalCp03-propeptide) showed a low nanomolar Ki value of 2.3 ± 0.2 nM against cathepsin L. A significant inhibition of tumor cell invasion was observed with H1975, HT29, MDA-BM-231, PANC1, and PC3 with a 76, 67, 67, 63, and 79% reduction, respectively, in invasion observed in the presence of 400 nM of the rSnuCalCp03-propeptide. In addition, thermal and pH study showed rSnuCalCp03-propeptide consisting of secondary structures was stable at a broad range of temperatures (30-70 °C) and pH (2-10, except for 5 which is close to the isoelectric point of 5.2).

Erythorbyl laurate as a potential food additive with multi-functionalities: Interfacial characteristics and antioxidant activity.

The interfacial characteristics and antioxidant activities of erythorbyl laurate were investigated to provide information on practical applications as a multi-functional food additive. The critical micelle concentration (CMC) of erythorbyl laurate was 0.101mM and its foam stability was three times (half-life 24.33±0.94h) higher than that of Tween 20 (8.00±1.63h). In free radical scavenging assay, the negligible decrease in EC50 of erythorbyl laurate compared to erythorbic acid manifested that C-5 selective esterification of erythorbic acid with an acyl group (lauric acid) did not reduce the inherent antioxidant activity of the donor (erythorbic acid). Erythorbyl laurate formed lipid peroxides slower (i.e. retarded oxidation) in an emulsion system than did erythorbic acid. The localization of erythorbyl laurate as an emulsifier allowed the antioxidant molecules to be concentrated at the oil-water interface where oxidation is prevalent, which led to more effective retardation of lipid oxidation.

Pseudonotohymena antarctica n. g., n. sp. (Ciliophora, Hypotricha), a New Species from Antarctic Soil.

A new soil ciliate, Pseudonotohymena antarctica n. g., n. sp., from King George Island, Antarctica, is described based on live observation, protargol impregnation, and its 18S rRNA gene. The new genus Pseudonotohymena is morphologically similar to the genus Notohymena Blatterer and Foissner in the following characteristics: 18 fronto-ventral-transverse cirri, a flexible body, undulating membranes, dorsomarginal kineties, and the number of cirri in the marginal rows. However, Pseudonotohymena differs from Notohymena particularly in the dorsal ciliature, that is, in possessing a nonfragmented dorsal kinety (vs. fragmented). In addition, the molecular phylogenetic relationship of the new species differs from that of Notohymena species. On the basis of the morphological features, the genetic data, and morphogenesis, we establish P. antarctica n. g., n. sp. In addition, the cyst morphology of this species is described.

Molecular Cloning and Expression Analysis of Pig Cd90.

The CD90 (Thy-1) is a glycosylphosphatidylinositol (GPI)-anchored glycoprotein that transfers signals involved in many biological events including cell activation, cell migration, cell adhesion, and tumor suppression. In this study, we cloned pig CD90 cDNA and determined its complete cDNA sequence. Pig CD90 cDNA contained an open reading frame (486 bp) encoding 161 amino acids with three putative N-glycosylation sites and four well-conserved cysteine residues, which form a possible disulfide bond within the extracellular domain among mammalian species. Pig CD90 mRNA was detected in various tissues, indicating the multicellular functions of CD90 in pigs. Flow cytometry analyses demonstrated that anti-human CD90 antibody recognizes a pig CD90 on the cell surface. Moreover, immunohistochemistry analysis revealed that CD90 expression is widely diffused in several pig tissues. Further studies will be necessary to define the functional contribution of CD90 during specific infectious diseases in pigs.

Glabretal-type triterpenoid from the root bark of Dictamnus dasycarpus ameliorates collagen-induced arthritis by inhibiting Erk-dependent lymphocyte proliferation.

ETHNOPHARMACOLOGICAL RELEVANCE: The root bark of Dictamnus dasycarpus Turcz. (Rutaceae) has been used as a traditional herbal medicine to treat various inflammatory diseases in East Asia. We have showed previously that a glabretal type triterpenoid (dictabretol A) from D. dasycarpus root bark has immunosuppressive activity. AIM OF THE STUDY: This study was conducted to define the molecular mechanism of how dictabretol A inhibits lymphocyte proliferation and to evaluate the therapeutic efficacy of dictabretol A in an animal model of rheumatoid arthritis. MATERIALS AND METHODS: Various murine immune cells (T cells, B cells, and macrophages) and splenocytes were used to study the anti-proliferative effect of dictabretol A in vitro. A collagen-induced arthritis model was also used to examine the therapeutic effect of dictabretol A in vivo. RESULTS: Dictabretol A specifically inhibited lymphocyte proliferation by blocking the cell cycle transition from the G1 to the S phase. This effect was achieved by blocking Erk1/2, nuclear factor kappa B, and the C-myc axis of cell cycle progression. Further dictabretol A treatment alleviated the severity of collagen-induced arthritis. CONCLUSION: Our results reveal the molecular mechanism for the anti-lymphoproliferative effect of dictabretol A and show the therapeutic efficacy of dictabretol A for rheumatoid arthritis.

Cysteine Protease Profiles of the Medicinal Plant Calotropis procera R. Br. revealed by de novo transcriptome analysis.

Calotropis procera R. Br., a traditional medicinal plant in India, is a promising source of commercial proteases, because the cysteine proteases from the plant exhibit high thermo-stability, broad pH optima, and plasma-clotting activity. Though several proteases such as Procerain, Procerain B, CpCp-1, CpCp-2, and CpCp-3 have been isolated and characterized, the information of their transcripts is limited to cDNAs encoding their mature peptides. Due to this limitation, in this study, to determine the cDNA sequences encoding full open reading frame of these cysteine proteases, transcripts were sequenced with an Illumina Hiseq2000 sequencer. A total of 171,253,393 clean reads were assembled into 106,093 contigs with an average length of 1,614 bp and an N50 of 2,703 bp, and 70,797 contigs with an average length of 1,565 bp and N50 of 2,082 bp using Trinity and Velvet-Oases software, respectively. Among these contigs, we found 20 unigenes related to papain-like cysteine proteases by BLASTX analysis against a non-redundant NCBI protein database. Our expression analysis revealed that the cysteine protease contains an N-terminal pro-peptide domain (inhibitor region), which is necessary for correct folding and proteolytic activity. It was evident that expression yields using an inducible T7 expression system in Escherichia coli were considerably higher with the pro-peptide domain than without the domain, which could contribute to molecular cloning of the Calotropis procera protease as an active form with correct folding.

Morphology, morphogenesis, and molecular phylogeny of Anteholosticha multicirrata n. sp. (Ciliophora, Spirotrichea) with a note on morphogenesis of A. pulchra (Kahl, 1932) Berger, 2003.

Two marine urostylid ciliates, Anteholosticha multicirrata n. sp. and Anteholosticha pulchra (Kahl, 1932) Berger, 2003, were collected from South Korea. These species were identified based on morphology, morphogenesis, and SSU rRNA gene sequence comparison. Anteholosticha multicirrata n. sp. is characterized by the following features: body size 90-125 × 30-45 µm in vivo, shape slender to ellipsoidal in outline, with yellow-greenish cortical granules distributed along and between dorsal kineties and cirri; single contractile vacuole positioned on left at mid-body; three frontal, five to seven frontoterminal, one buccal, one to two pretransverse and four to six transverse cirri; three complete dorsal kineties; one left and one right marginal cirral row; about 117 macronuclear nodules; and three to four micronuclei observed during morphogenesis. In addition, based on the observations of morphogenesis, we found that A. pulchra has pretransverse cirri, which were not described in detail in previous studies. Nuclear small subunit ribosomal RNA (SSU rRNA) gene was used to analyse their phylogenetic relationship, and the gene tree supports that the genus Anteholosticha is a highly polyphyletic group.

Inhibitory effects of arbutin-beta-glycosides synthesized from enzymatic transglycosylation for melanogenesis.

To develop a new skin whitening agent, arbutin-beta-glycosides were synthesized and evaluated for their melanogenesis inhibitory activities. Three active compounds were synthesized via the transglycosylation reaction of Thermotoga neapolitana beta-glucosidase and purified by recycling preparative HPLC. As compared with arbutin (IC(50 )= 6 mM), the IC(50 )values of these compounds were 8, 10, and 5 mM for beta-D -glucopyranosyl-(1-->6)-arbutin, beta-D: -glucopyranosyl-(1-->4)-arbutin, and beta-D -glucopyranosyl-(1-->3)-arbutin, respectively. beta-D: -Glucosyl-(1-->3)-arbutin also exerted the most profound inhibitory effects on melanin synthesis in B16F10 melanoma cells. Melanin synthesis was inhibited to a significant degree at 5 mM, at which concentration the melanin content was reduced to below 70% of that observed in the untreated cells. Consequently, beta-D: -glucopyranosyl-(1-->3)-arbutin is a more effective depigmentation agent and is also less cytotoxic than the known melanogenesis inhibitor, arbutin.