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1.
Chemosphere ; 361: 142430, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38844105

ABSTRACT

In the present study, algicidal bacteria cultivated in an aqueous medium were utilized as a surface modification agent to develop an efficient adsorbent for the removal of Microcystis aeruginosa. The modification considerably enhanced M. aeruginosa cell removal efficiency. Moreover, the introduction of bio-compounds ensured specificity in the removal of M. aeruginosa. Additionally, the cyanotoxin release and acute toxicity tests demonstrated that the adsorption process using the developed adsorbent is environmentally safe. Furthermore, the practical feasibility of the adsorptive removal of M. aeruginosa was confirmed through cell removal tests performed using the developed adsorbent in a scaled-up reactor (50 L and 10 tons). In these tests, the effects of the adsorbent application type, water temperature, and initial cell concentration on the M. aeruginosa removal efficiency were evaluated. The results of this study provide novel insights into the valorization strategy of biological algicides repurposed as adsorbents, and provide practical operational data for effective M. aeruginosa removal in scaled-up conditions.


Subject(s)
Microcystis , Adsorption , Microcystins/chemistry , Microcystins/metabolism , Microcystins/isolation & purification , Cyanobacteria/metabolism , Water Purification/methods
2.
J Vet Sci ; 25(2): e22, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38568824

ABSTRACT

BACKGROUND: Achilles tendon is composed of dense connective tissue and is one of the largest tendons in the body. In veterinary medicine, acute ruptures are associated with impact injury or sharp trauma. Healing of the ruptured tendon is challenging because of poor blood and nerve supply as well as the residual cell population. Platelet-rich plasma (PRP) contains numerous bioactive agents and growth factors and has been utilized to promote healing in bone, soft tissue, and tendons. OBJECTIVE: The purpose of this study was to evaluate the healing effect of PRP injected into the surrounding fascia of the Achilles tendon after allograft in rabbits. METHODS: Donor rabbits (n = 8) were anesthetized and 16 lateral gastrocnemius tendons were fully transected bilaterally. Transected tendons were decellularized and stored at -80°C prior to allograft. The allograft was placed on the partially transected medial gastrocnemius tendon in the left hindlimb of 16 rabbits. The allograft PRP group (n = 8) had 0.3 mL of PRP administered in the tendon and the allograft control group (n = 8) did not receive any treatment. After 8 weeks, rabbits were euthanatized and allograft tendons were transected for macroscopic, biomechanical, and histological assessment. RESULTS: The allograft PRP group exhibited superior macroscopic assessment scores, greater tensile strength, and a histologically enhanced healing process compared to those in the allograft control group. CONCLUSIONS: Our results suggest administration of PRP on an allograft tendon has a positive effect on the healing process in a ruptured Achilles tendon.


Subject(s)
Achilles Tendon , Platelet-Rich Plasma , Tendon Injuries , Rabbits , Animals , Achilles Tendon/surgery , Achilles Tendon/injuries , Achilles Tendon/pathology , Tendon Injuries/therapy , Tendon Injuries/veterinary , Tendon Injuries/pathology , Wound Healing , Allografts/pathology
3.
Article in English | MEDLINE | ID: mdl-38376819

ABSTRACT

Human intestinal epithelial cells (IECs) play an important role in maintaining gut homeostasis by producing antimicrobial peptides (AMPs). Bacillus subtilis, a commensal bacterium, is considered a probiotic. Although its protective effects on intestinal health are widely reported, the key component of B. subtilis responsible for its beneficial effects remains elusive. In this study, we tried to identify the key molecules responsible for B. subtilis-induced AMPs and their molecular mechanisms in a human IEC line, Caco-2. B. subtilis increased human beta defensin (HBD)-2 mRNA expression in a dose- and time-dependent manner. Among the B. subtilis microbe-associated molecular patterns, lipoprotein (LPP) substantially increased the mRNA expression and protein production of HBD-2, whereas lipoteichoic acid and peptidoglycan did not show such effects. Those results were confirmed in primary human IECs. In addition, both LPP recognition and HBD-2 secretion mainly took place on the apical side of fully differentiated and polarized Caco-2 cells through Toll-like receptor 2-mediated JNK/p38 MAP kinase/AP-1 and NF-κB pathways. HBD-2 efficiently inhibited the growth of the intestinal pathogens Staphylococcus aureus and Bacillus cereus. Furthermore, LPPs pre-incubated with lipase or proteinase K decreased LPP-induced HBD-2 expression, suggesting that the lipid and protein moieties of LPP are crucial for HBD-2 expression. Q Exactive Plus mass spectrometry identified 35 B. subtilis LPP candidates within the LPP preparation, and most of them were ABC transporters. Taken together, these results suggest that B. subtilis promotes HBD-2 secretion in human IECs mainly with its LPPs, which might enhance the protection from intestinal pathogens.

4.
Vet Sci ; 10(11)2023 Oct 27.
Article in English | MEDLINE | ID: mdl-37999457

ABSTRACT

Intra-abdominal pressure (IAP) elevation during capnoperitoneum can cause adverse cardiovascular and respiratory effects. This study aimed to determine if a sequentially increased IAP affects cardiovascular and respiratory variables in anesthetized dogs and evaluate the effects of the constant-rate infusion of dexmedetomidine (Dex) on cardiovascular and respiratory variables with increased IAP. Five dogs were anesthetized and instrumented, and a Veress needle was equipped to adjust the IAP using a carbon dioxide insufflator. Stabilization was conducted for 1 h, and physiological variables were measured at IAPs of 0, 5, 10, 15, and 20 mmHg and after desufflation. After the washout period, the dogs underwent similar procedures along with a constant-rate infusion of dexmedetomidine. The cardiovascular effects of increased IAP up to 20 mmHg were not significant in healthy beagle dogs and those administered with dexmedetomidine. When comparing the control and dexmedetomidine groups, the overall significant effects of dexmedetomidine were noted on heart rate, cardiac output, and systemic vascular resistance during the experiment. Respiratory effects were not observed during abdominal insufflation when compared between different IAPs and between the two groups. Overall, an increased IAP of up to 20 mmHg did not significantly affect cardiovascular and respiratory variables in both the control and dexmedetomidine groups. This study suggests that the administration of a dexmedetomidine infusion is applicable in laparoscopic procedures in healthy dogs.

5.
Med Phys ; 50(12): 7498-7512, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37669510

ABSTRACT

BACKGROUND: The bowtie-filter in cone-beam CT (CBCT) causes spatially nonuniform x-ray beam often leading to eclipse artifacts in the reconstructed image. The artifacts are further confounded by the patient scatter, which is therefore patient-dependent as well as system-specific. PURPOSE: In this study, we propose a dual-domain network for reducing the bowtie-filter-induced artifacts in CBCT images. METHODS: In the projection domain, the network compensates for the filter-induced beam-hardening that are highly related to the eclipse artifacts. The output of the projection-domain network was used for image reconstruction and the reconstructed images were fed into the image-domain network. In the image domain, the network further reduces the remaining cupping artifacts that are associated with the scatter. A single image-domain-only network was also implemented for comparison. RESULTS: The proposed approach successfully enhanced soft-tissue contrast with much-reduced image artifacts. In the numerical study, the proposed method decreased perceptual loss and root-mean-square-error (RMSE) of the images by 84.5% and 84.9%, respectively, and increased the structure similarity index measure (SSIM) by 0.26 compared to the original input images on average. In the experimental study, the proposed method decreased perceptual loss and RMSE of the images by 87.2% and 92.1%, respectively, and increased SSIM by 0.58 compared to the original input images on average. CONCLUSIONS: We have proposed a deep-learning-based dual-domain framework to reduce the bowtie-filter artifacts and to increase the soft-tissue contrast in CBCT images. The performance of the proposed method has been successfully demonstrated in both numerical and experimental studies.


Subject(s)
Neural Networks, Computer , Quality Improvement , Humans , Cone-Beam Computed Tomography/methods , Image Processing, Computer-Assisted/methods , X-Rays , Algorithms , Phantoms, Imaging , Artifacts
6.
Animals (Basel) ; 12(21)2022 Oct 28.
Article in English | MEDLINE | ID: mdl-36359090

ABSTRACT

The release of reproductive hormones in the hypothalamic-pituitary-gonadal (HPG) axis is regulated by its upstream regulator, kisspeptin, and influenced by external stresses, including heat stress. Since the effect of heat stress (summer infertility) on hypothalamic kisspeptin expression in domestic sows is not yet understood, the present study attempted to identify changes in kisspeptin expression in different seasons (summer and spring). The high atmospheric temperature in summer decreased the pregnancy rate and litter size and increased stress-related hormones as a chronic stressor to domestic sows. The hypothalamic kisspeptin expression in summer was decreased regardless of the estrus phase and negatively correlated with atmospheric temperature, indicating that high temperature decreased kisspeptin. When the activity of hypothalamic kisspeptin neurons in the follicular phase was assessed using c-Fos staining, a decreased number of kisspeptin neurons coexpressing c-Fos was observed in domestic sows in summer. Accordingly, lower expression of kisspeptin induced decreased levels of HPG axis-related reproductive hormones, such as gonadotropins and estrogen, and fewer large ovarian follicles. In conclusion, the present study demonstrated that reduced kisspeptin expression and its neuronal activity in the hypothalamus under heat stress in summer induced downregulation of the HPG axis and caused summer infertility in domestic sows.

8.
Anim Biosci ; 35(12): 1850-1859, 2022 Dec.
Article in English | MEDLINE | ID: mdl-35507843

ABSTRACT

OBJECTIVE: The quantitative reverse transcription polymerase chain reaction (qPCR) is the most accurate and reliable technique for analysis of gene expression. Endogenous reference genes (RGs) have been used to normalize qPCR data, although their expression may vary in different tissues and experimental conditions. Verification of the stability of RGs in selected samples is a prerequisite for reliable results. Therefore, we attempted to identify the most stable RGs in the hypothalamic-pituitary-gonadal (HPG) axis in sows. METHODS: The cycle threshold values of nine commonly used RGs (18S, HPRT1, GAPDH, RPL4, PPIA, B2M, YWHAZ, ACTB, and SDHA) from HPG axis-related tissues in the domestic sows in the different stages of estrus cycle were analyzed using two RG-finding programs, geNorm and Normfinder, to rank the stability of the pool of RGs. In addition, the effect of the most and least stable RGs was examined by normalization of the target gene, gonadotropin-releasing hormone (GnRH), in the hypothalamus. RESULTS: PPIA, HPRT1, and YWHAZ were the most stable RGs in the HPG axis-related tissues in sows regardless of the stages of estrus cycle. In contrast, traditional RGs, including 18S and ACTB, were found to be the least stable under these experimental conditions. In particular, in the normalization of GnRH expression in the hypothalamus against several stable RGs, PPIA, HPRT1, and YWHAZ, could generate significant (p<0.05) elevation of GnRH in the preovulatory phase compared to the luteal phase, but the traditional RGs with the least stability (18S and ACTB) did not show a significant difference between groups. CONCLUSION: These results indicate the importance of verifying RG stability prior to commencing research and may contribute to experimental design in the field of animal reproductive physiology as reference data.

9.
J Xray Sci Technol ; 30(3): 549-566, 2022.
Article in English | MEDLINE | ID: mdl-35253722

ABSTRACT

BACKGROUND: Dual-energy computed tomography (DECT) is a widely used and actively researched imaging modality that can estimate the physical properties of an object more accurately than single-energy CT (SECT). Recently, iterative reconstruction methods called one-step methods have received attention among various approaches since they can resolve the intermingled limitations of the conventional methods. However, the one-step methods typically have expensive computational costs, and their material decomposition performance is largely affected by the accuracy in the spectral coefficients estimation. OBJECTIVE: In this study, we aim to develop an efficient one-step algorithm that can effectively decompose into the basis material maps and is less sensitive to the accuracy of the spectral coefficients. METHODS: By use of a new loss function that employs the non-linear forward model and the weighted squared errors, we propose a one-step reconstruction algorithm named generalized simultaneous algebraic reconstruction technique (GSART). The proposed algorithm was compared with the image-domain material decomposition and other existing one-step reconstruction algorithm. RESULTS: In both simulation and experimental studies, we demonstrated that the proposed algorithm effectively reduced the beam-hardening artifacts thereby increasing the accuracy in the material decomposition. CONCLUSIONS: The proposed one-step reconstruction for material decomposition in dual-energy CT outperformed the image-domain approach and the existing one-step algorithm. We believe that the proposed method is a practically very useful addition to the material-selective image reconstruction field.

10.
Differentiation ; 125: 18-26, 2022.
Article in English | MEDLINE | ID: mdl-35349880

ABSTRACT

Mouse embryonic stem cells (mESCs) are characterized by self-renewal and pluripotency and can undergo differentiation into the three germ layers (ectoderm, mesoderm, and endoderm). Melanoma-associated antigen D1 (Maged1), which is expressed in all developing and adult tissues, modulates tissue regeneration and development. In the present study, we examined the expression and function of Maged1 in mESCs. Maged1 protein and mRNA expression increased during mESC differentiation. The pluripotency of mESCs was significantly reduced through extracellular signal-regulated kinase 1/2 phosphorylation upon knockdown of Maged1, and through G1 cell cycle arrest during cell division, resulting in significantly reduced mESC proliferation. Moreover, the diameter of the embryoid bodies was significantly reduced, accompanied by increased levels of ectodermal differentiation markers and decreased levels of mesodermal and endodermal differentiation markers. Maged1-knockdown mESC lines showed significantly reduced teratoma volumes and inhibition of teratoma formation in nude mice. Additionally, we observed increased ectodermal markers but decreased mesodermal and endodermal markers in teratoma tissues. These findings show that Maged1 affects mESC pluripotency, proliferation, cell cycle, and differentiation, thereby contributing to our understanding of the basic molecular biological mechanisms and potential roles of Maged1 as a regulator of various mESC properties.


Subject(s)
Mouse Embryonic Stem Cells , Animals , Antigens, Differentiation/metabolism , Cell Cycle/genetics , Cell Death , Cell Differentiation/genetics , Cell Division , Mice , Mice, Nude , Mouse Embryonic Stem Cells/cytology , Mouse Embryonic Stem Cells/metabolism , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Teratoma/genetics , Teratoma/metabolism , Teratoma/pathology
11.
Microbiol Spectr ; 10(1): e0161421, 2022 02 23.
Article in English | MEDLINE | ID: mdl-35171037

ABSTRACT

The antigen-based rapid diagnostic test (Ag-RDT) using saliva specimens is fast, noninvasive, and suitable for SARS-CoV-2 self-testing, unlike nasopharyngeal swab (NPS) testing. We evaluated a novel Beanguard gargle (BG)-based virus collection method that can be applied to Ag-RDT as an alternative to the current RT-PCR with an NPS for early diagnosis of COVID-19. This clinical trial comprised 102 COVID-19-positive patients hospitalized after a governmental screening process and 100 healthy individuals. Paired NPS and BG-based saliva specimens from COVID-19 patients and healthy individuals were analyzed using NPS-RT-PCR, BG-RT-PCR, and BG-Ag-RDTs, whose diagnostic performance for detecting SARS-CoV-2 was compared. BG-Ag-RDTs showed high sensitivity (97.8%) and specificity (100%) in 45 patients within 6 days of illness and detected all cases of SARS-CoV-2 Alpha and Delta variants. In 11 asymptomatic active COVID-19 cases, both BG-Ag-RDTs and BG-RT-PCR showed sensitivities and specificities of 100%. Sensitivities of BG-Ag-RDT and BG-RT-PCR toward salivary viral detection were highly concordant, with no discrimination between symptomatic (97.0%), asymptomatic (100%), or SARS-CoV-2 variant (100%) cases. The intermolecular interactions between SARS-CoV-2 spike proteins and truncated canavalin, an active ingredient from the bean extract (BE), were observed in terms of physicochemical properties. The detachment of the SARS-CoV-2 receptor-binding domain from hACE2 increased as the BE concentration increased, allowing the release of the virus from hACE2 for early diagnosis. Using BG-based saliva specimens remarkably enhances the Ag-RDT diagnostic performance as an alternative to NPS and enables noninvasive, rapid, and accurate COVID-19 self-testing and mass screening, supporting efficient COVID-19 management. IMPORTANCE An Ag-RDT is less likely to be accepted as an initial test method for early diagnosis owing to its low sensitivity. However, our self-collection method, Ag-RDT using BG-based saliva specimens, showed significantly enhanced detection sensitivity and specificity toward SARS-CoV-2 including the Alpha and Delta variants in all patients tested within 6 days of illness. The method represents an attractive alternative to nasopharyngeal swabs for the early diagnosis of symptomatic and asymptomatic COVID-19 cases. The evidence suggests that the method could have a potential for mass screening and monitoring of COVID-19 cases.


Subject(s)
COVID-19 Serological Testing/methods , COVID-19/diagnosis , SARS-CoV-2/isolation & purification , Saliva/virology , Adult , Aged , Aged, 80 and over , COVID-19/virology , COVID-19 Nucleic Acid Testing , COVID-19 Serological Testing/instrumentation , Female , Humans , Male , Middle Aged , Nasopharynx/virology , Republic of Korea , SARS-CoV-2/genetics , SARS-CoV-2/immunology , Sensitivity and Specificity , Young Adult
12.
Life Sci ; 288: 120170, 2022 Jan 01.
Article in English | MEDLINE | ID: mdl-34826438

ABSTRACT

AIMS: Antitumor effects of veratramine in prostate and liver cancers has been investigated, but it is still unclear whether veratramine can be used as an effective therapeutic agent for glioma. The aim of this study was to evaluate the potential pharmacological mechanism of veratramine in glioma. MAIN METHODS: Using four types of human glioblastoma cell lines, including A172, HS-683, T98G, and U-373-MG the dose-dependent antitumor effect of veratramine was evaluated. The cytotoxicity and cell proliferation were examined by CCK-8, and cell proliferation was further confirmed by anchorage-independent colony formation assay. The cell cycle distribution and apoptotic rate was assessed by flow cytometry, and apoptosis was further evaluated by apoptosis assay. The migration and invasiveness capacity were analyzed by using transwell. Protein and mRNA levels of related factors were determined by western blotting and RT-qPCR, respectively. KEY FINDINGS: Veratramine markedly induced apoptosis, suppressed the cell proliferation via the cell cycle G0/G1 phase arrest, and reduced the capacity for the migration and invasion in human glioblastoma multiforme cell lines. Moreover, veratramine was sufficient to affect the phosphatidylinositol-3-kinase/serine-threonine kinase/mechanistic target of rapamycin signaling pathway and its downstream Mdm2/p53/p21 pathway in human glioblastoma cell lines. SIGNIFICANCE: Antitumor effects of veratramine in suppression of glioma progression was mediated by the regulation of PI3K/Akt/mTOR and Mdm2/p53/p21 signaling pathway.


Subject(s)
Gene Expression Regulation, Neoplastic/drug effects , Glioblastoma/drug therapy , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , TOR Serine-Threonine Kinases/metabolism , Veratrum Alkaloids/pharmacology , Apoptosis , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Cell Cycle , Cell Movement , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p21/genetics , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Glioblastoma/genetics , Glioblastoma/metabolism , Glioblastoma/pathology , Humans , Neoplasm Invasiveness , Phosphatidylinositol 3-Kinases/genetics , Proto-Oncogene Proteins c-akt/genetics , Proto-Oncogene Proteins c-mdm2/genetics , Proto-Oncogene Proteins c-mdm2/metabolism , TOR Serine-Threonine Kinases/genetics , Tumor Cells, Cultured , Tumor Suppressor Protein p53/genetics , Tumor Suppressor Protein p53/metabolism
13.
Appl Microbiol Biotechnol ; 106(2): 663-673, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34971409

ABSTRACT

For the efficient production of heterologous proteins in the yeast Saccharomyces cerevisiae, we screened for a novel fusion partner from the yeast secretome. From twenty major proteins identified from the yeast secretome, we selected Scw4p, a cell wall protein with similarity to glucanase, and modified to develop a general fusion partner for the secretory expression of heterologous proteins in yeast. The optimal size of the SCW4 gene to act as an efficient fusion partner was determined by C-terminal truncation analysis; two of the variants, S1 (truncated at codon 115Q) and S2 (truncated at codon 142E), were further used for the secretion of heterologous proteins. When fused with S2, the secretion of three target proteins (hGH, exendin-4, and hPTH) significantly increased. Conserved O-glycosylation sites (Ser/Thr-rich domain) and hydrophilic sequences of S2 were deemed important for the function of S2 as a secretion fusion partner. Approximately 5 g/L of the S2-exendin-4 fusion protein was obtained from fed-batch fermentation. Intact target proteins were easily purified by affinity chromatography after in vitro processing of the fusion partner. This system may be of general application for the secretory production of heterologous proteins in S. cerevisiae. KEY POINTS : • Target proteins were efficiently secreted with their N-terminus fused to Scw4p. • O-glycosylation and hydrophilic stretches in Scw4p were important for protein secretion. • A variant of Scw4p (S2) was successfully applied for the secretory expression of heterologous proteins.


Subject(s)
Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism , Secretome
14.
J Exp Clin Cancer Res ; 40(1): 291, 2021 Sep 18.
Article in English | MEDLINE | ID: mdl-34537073

ABSTRACT

BACKGROUND: The progression of prostate cancer (PC) to the highly aggressive metastatic castration-resistant prostate cancer (mCRPC) or neuroendocrine prostate cancer (NEPC) is a fatal condition and the underlying molecular mechanisms are poorly understood. Here, we identified the novel transcriptional factor ZNF507 as a key mediator in the progression of PC to an aggressive state. METHODS: We analyzed ZNF507 expression in the data from various human PC database and high-grade PC patient samples. By establishment of ZNF507 knockdown and overexpression human PC cell lines, we assessed in vitro PC phenotype changes including cell proliferation, survival, migration and invasion. By performing microarray with ZNF507 knockdown PC cells, we profiled the gene clusters affected by ZNF507 knockdown. Moreover, ZNF507 regulated key signal was evaluated by dual-luciferase reporter and chromatin immunoprecipitation (ChIP) assays. Finally, we performed xenograft and in vivo metastasis assay to confirm the effect of ZNF507 knockdown in PC cells. RESULTS: We found that ZNF507 expression was increased, particularly in the highly graded PC. ZNF507 was also found to be associated with metastatic PC of a high grade. Loss- or gain-of-function-based analysis revealed that ZNF507 promotes the growth, survival, proliferation, and metastatic properties of PC (e.g., epithelial-mesenchymal transition) by upregulating TGF-ß signaling. Profiling of gene clusters affected by ZNF507 knockdown revealed that ZNF507 positively regulated the transcription of TGFBR1, MAP3K8, and FURIN, which in turn promoted the progression of PC to highly metastatic and aggressive state. CONCLUSIONS: Our findings suggest that ZNF507 is a novel key regulator of TGF-ß signaling in the progression of malignant PC and could be a promising target for studying the development of advanced metastatic PCs.


Subject(s)
DNA-Binding Proteins/metabolism , MAP Kinase Kinase Kinases/metabolism , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/pathology , Proto-Oncogene Proteins/metabolism , Receptor, Transforming Growth Factor-beta Type I/metabolism , Signal Transduction , Transforming Growth Factor beta/metabolism , Animals , Apoptosis/genetics , Biomarkers , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation , DNA-Binding Proteins/genetics , Disease Progression , Disease Susceptibility , Gene Expression , Gene Knockdown Techniques , Heterografts , Humans , Male , Mice , Models, Biological , Prognosis , Prostatic Neoplasms/etiology
15.
Vet Sci ; 8(9)2021 Sep 03.
Article in English | MEDLINE | ID: mdl-34564576

ABSTRACT

Adipose tissue-derived mesenchymal stem cells (AD-MSCs) release extracellular vesicles such as exosomes, apoptotic bodies, and microparticles. In particular, exosomes are formed inside cells via multivesicular bodies (MVBs), thus their protein, DNA, and RNA content are similar to those of the parent cells. Exosome research is rapidly expanding, with an increase in the number of related publications observed in recent years; therefore, the function and application of MSC-derived exosomes could emerge as cell-free therapeutics. Exosomes have been isolated from feline AD-MSCs and feline fibroblast cell culture media using ultracentrifugation. Feline exosomes have been characterized by FACS, nanoparticle tracking analysis, and transmission electron microscopy imaging. Moreover, cytokine levels were detected by sandwich enzyme-linked immunosorbent assay in exosomes and LPS-induced THP-1 macrophages. The size of the isolated exosomes was that of a typical exosome, i.e., approximately 150 nm, and they expressed tetraspanins CD9 and CD81. The anti-inflammatory factor IL-10 was increased in feline AD-MSC-derived exosomes. However, pro-inflammatory factors such as IL-1ß, IL-8, IL-2, RANTES, and IFN-gamma were significantly decreased in feline AD-MSC-derived exosomes. This was the first demonstration that feline AD-MSC-derived exosomes enhance the inflammatory suppressive effects and have potential for the treatment of immune diseases or as an inflammation-inhibition therapy.

16.
Biomed Opt Express ; 12(8): 4837-4851, 2021 Aug 01.
Article in English | MEDLINE | ID: mdl-34513228

ABSTRACT

Diffuse optical tomography (DOT) is a non-invasive functional imaging modality that uses near-infrared (NIR) light to measure the oxygenation state and the concentration of hemoglobin. By complementarily using DOT with other anatomical imaging modalities, physicians can diagnose more accurately through additional functional image information. In breast imaging, diagnosis of dense breasts is often challenging because the bulky fibrous tissues may hinder the correct tumor characterization. In this work, we proposed a three-compartment-breast (3CB) decomposition-based prior-guided optical tomography for enhancing DOT image quality. We conjectured that the 3CB prior would lead to improvement of the spatial resolution and also of the contrast of the reconstructed tumor image, particularly for the dense breasts. We conducted a Monte-Carlo simulation to acquire dual-energy X-ray projections of a realistic 3D numerical breast phantom and performed digital breast tomosynthesis (DBT) for setting up a 3CB model. The 3CB prior was then used as a structural guide in DOT image reconstruction. The proposed method resulted in the higher spatial resolution of the recovered tumor even when the tumor is surrounded by the fibroglandular tissues compared with the typical two-composition-prior method or the standard Tikhonov regularization method.

17.
Biomed Res Int ; 2021: 4604856, 2021.
Article in English | MEDLINE | ID: mdl-34527737

ABSTRACT

IFN-γ licensing to mesenchymal stem cells (MSCs) is applied to enhance the therapeutic potential of MSCs. However, although the features of MSCs are affected by several stimuli, little information is available on changes to the therapeutic potential of IFN-γ-licensed differentiated MSCs during xenogeneic applications. Therefore, the present study is aimed at clarifying the effects of adipogenic/osteogenic differentiation and IFN-γ licensing on the in vitro immunomodulatory and migratory properties of porcine bone marrow-derived MSCs in xenogeneic applications using human peripheral blood mononuclear cells (PBMCs). IFN-γ licensing in differentiated MSCs lowered lineage-specific gene expression but did not affect MSC-specific cell surface molecules. Although indoleamine 2,3 deoxygenase (IDO) activity and expression were increased after IFN-γ licensing in undifferentiated MSCs, they were reduced after differentiation. IFN-γ licensing to differentiated MSCs elevated the reduced IDO expression in differentiated MSCs; however, the increase was not sufficient to reach to the level achieved by undifferentiated MSCs. During a mixed lymphocyte reaction with quantification of TNF-α concentration, proliferation and activation of xenogeneic PBMCs were suppressed by undifferentiated MSCs but inhibited to a lesser extent by differentiated MSCs. IFN-γ licensing increasingly suppressed proliferation of PBMCs in undifferentiated MSCs but it was incapable of elevating the reduced immunosuppressive ability of differentiated MSCs. Migratory ability through a scratch assay and gene expression study was reduced in differentiated MSCs than their undifferentiated counterparts; IFN-γ licensing was unable to enhance the reduced migratory ability in differentiated MSCs. Similar results were found in a Transwell system with differentiated MSCs in the upper chamber toward xenogeneic PBMCs in the lower chamber, despite IFN-γ licensing increased the migratory ability of undifferentiated MSCs. Overall, IFN-γ licensing did not enhance the reduced immunomodulatory and migratory properties of differentiated MSCs in a xenogeneic application. This study provides a better understanding of the ways in which MSC therapy can be applied.


Subject(s)
Interferon-gamma/metabolism , Mesenchymal Stem Cell Transplantation/methods , Mesenchymal Stem Cells/metabolism , Animals , Bone Marrow/metabolism , Bone Marrow Cells/metabolism , Cell Differentiation/drug effects , Cell Movement/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Coculture Techniques , Cytokines/metabolism , Heterografts/metabolism , Humans , Immunomodulation/drug effects , Interferon-gamma/physiology , Mesenchymal Stem Cells/physiology , Osteogenesis/drug effects , Swine , Tumor Necrosis Factor-alpha/metabolism
18.
Int J Mol Sci ; 22(5)2021 Feb 26.
Article in English | MEDLINE | ID: mdl-33652881

ABSTRACT

Senescence marker protein 30 (SMP30) is a cell survival factor playing an important role in vitamin C synthesis and antiapoptosis. Moreover, its cytoprotective role suggests a possibility to be related to cancer cell survival. Mammary carcinoma is a common cancer in both humans and animals. Because of its histopathological diversity, especially in the early stage, histopathological diagnosis may be complicated; therefore, a diagnostic marker is helpful for confirmation. The present study analyzed the expression pattern of SMP30 in mammary carcinoma in humans, dogs, and cats. Immunohistochemistry, immunofluorescence, and western blot analysis were used to investigate SMP30 expression patterns. The expression was specifically observed in neoplastic glandular epithelial cells. The expression increased with the malignancy of glandular epithelial cells with a highly proliferative status. However, SMP30 expression was low in normal mammary gland tissues or well-differentiated adenoma tissues. The patterns were consistently reproduced in canine primary mammary carcinoma cells and MCF-7 and MDA-MB-231 human carcinoma cell lines. This study provides useful information to understand SMP30 expression in various stages of mammary carcinoma and to suggest its utility as a pan-species diagnostic marker, thereby helping to establish strategies for diagnosing mammary carcinoma in several species.


Subject(s)
Breast Neoplasms/pathology , Calcium-Binding Proteins/analysis , Cat Diseases/pathology , Dog Diseases/pathology , Intracellular Signaling Peptides and Proteins/analysis , Mammary Neoplasms, Animal/pathology , Animals , Biomarkers, Tumor/analysis , Breast/pathology , Breast Neoplasms/diagnosis , Cat Diseases/diagnosis , Cats , Cell Line, Tumor , Dog Diseases/diagnosis , Dogs , Female , Humans , MCF-7 Cells , Mammary Neoplasms, Animal/diagnosis , Prognosis
19.
Vet Sci ; 8(2)2021 Feb 07.
Article in English | MEDLINE | ID: mdl-33562192

ABSTRACT

Wharton's jelly is a well-known mesenchymal stem cell source in many species, including humans. However, there have been no reports confirming the presence of mesenchymal stem cells in Wharton's jelly in cats. The purpose of this study was to isolate mesenchymal stem cells (MSCs) from the Wharton's jelly of cats and to characterize stem cells. In this study, feline Wharton's jelly-derived mesenchymal stem cells (fWJ-MSCs) were isolated and successfully cultured. fWJ-MSCs were maintained and the proliferative potential was measured by cumulative population doubling level (CPDL) test, scratch test, and colony forming unit (CFU) test. Stem cell marker, karyotyping and immunophenotyping analysis by flow cytometry showed that fWJ-MSCs possessed characteristic mesenchymal stem cell markers. To confirm the differentiation potential, we performed osteogenic, adipogenic and chondrogenic induction under each differentiation condition. fWJ-MSCs has the ability to differentiate into multiple lineages, including osteogenic, adipogenic and chondrogenic differentiation. This study shows that Wharton's jelly of cat can be a good source of mesenchymal stem cells. In addition, fWJ-MSCs may be useful for stem cell-based therapeutic applications in feline medicine.

20.
J Proteome Res ; 19(1): 391-400, 2020 01 03.
Article in English | MEDLINE | ID: mdl-31689115

ABSTRACT

Motile cilia of multiciliated epithelial cells have important roles in animal development and cell homeostasis. Although several studies have identified and reported proteins localized in this complex organelle and the related immotile primary cilia from various cell types, it is still challenging to isolate high quantities of ciliary proteins for proteomic analysis. In this study, African clawed frog (Xenopus laevis) embryos, which have many multiciliated cells in the epidermis, were treated with a simple ionic buffer to identify 1009 proteins conserved across vertebrates; these proteins were putatively localized in motile cilia. Using two ciliary proteome databases, we confirmed that previously validated cilia-associated proteins are highly enriched in our ciliary proteome. Proteins localized at the transition zone and Ellis-van Creveld zone, which are distinct regions at the base of cilia, near the junction with the apical cell surface, were isolated using our method. Among the newly identified ciliary proteins, we report that KRT17 may have an unrecognized function in motile cilia. Hence, the method developed in this study would be useful for understanding the ciliary proteome.


Subject(s)
Cilia/metabolism , Keratin-17/metabolism , Proteomics/methods , Xenopus Proteins/analysis , Animals , Cilia/physiology , Embryo, Nonmammalian/cytology , Epidermis/metabolism , Female , Keratin-17/genetics , Male , Reproducibility of Results , Xenopus/embryology , Xenopus Proteins/metabolism , Xenopus laevis/embryology
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