A potential role of protein extractions from Metagonimus yokogawai in amelionating inflammation in patients with ankylosing spondylitis.

Helminth infections and their components has been recognized to have a positive impact on the immune system. This study aimed to investigate the potential of Metagonimus yokogawai-derived proteins (MYp) to provide protection against ankylosing spondylitis (AS) through modulation of immune responses. The cytotoxicity of MYp at various doses was first assessed using MTS and flow cytometry. Peripheral blood mononuclear cells (PBMCs) were collected from AS patients, and the production of inflammatory cytokines was analyzed through flow cytometry. In the experiments with SKG mice, MYp or vehicle was administered and inflammation was evaluated through immunohistochemistry and enzyme-linked immunosorbent assay. The results showed that MYp did not decrease cell viability of PBMCs even after 48 h. Additionally, the frequencies of IFN-γ and IL-17A producing cells were significantly reduced after MYp treatment in the PBMC cultures. Furthermore, MYp treatment significantly suppressed arthritis and enthesitis in the SKG mouse model. The results suggest the first evidence that MYp can effectively alleviate clinical symptoms and restore cytokine balance in patients with AS.

Breathing-Driven Self-Powered Pyroelectric ZnO Integrated Face Mask for Bioprotection.

Rapid spread of infectious diseases is a global threat and has an adverse impact on human health, livelihood, and economic stability, as manifested in the ongoing coronavirus disease 2019 (COVID-19) pandemic. Even though people wear a face mask as protective equipment, direct disinfection of the pathogens is barely feasible, which thereby urges the development of biocidal agents. Meanwhile, repetitive respiration generates temperature variation wherein the heat is regrettably wasted. Herein, a biocidal ZnO nanorod-modified paper (ZNR-paper) composite that is 1) integrated on a face mask, 2) harvests waste breathing-driven thermal energy, 3) facilitates the pyrocatalytic production of reactive oxygen species (ROS), and ultimately 4) exhibits antibacterial and antiviral performance is proposed. Furthermore, in situ generated compressive/tensile strain of the composite by being attached to a curved mask is investigated for high pyroelectricity. The anisotropic ZNR distortion in the bent composite is verified with changes in ZnO bond lengths and OZnO bond angles in a ZnO4 tetrahedron, resulting in an increased polarization state and possibly contributing to the following pyroelectricity. The enhanced pyroelectric behavior is demonstrated by efficient ROS production and notable bioprotection. This study exploring the pre-strain effect on the pyroelectricity of ZNR-paper might provide new insights into the piezo-/pyroelectric material-based applications.

CCL20 inhibition for treating inflammation in ankylosing spondylitis.

OBJECTIVES: Th17 cells are known to play a significant role in AS. C-C motif chemokine ligand 20 (CCL20) binds to C-C chemokine receptor 6 (CCR6) on Th17 cells, promoting their migration to inflammation sites. The aim of this research is to examine the effectiveness of CCL20 inhibition in treating inflammation in AS. METHODS: Mononuclear cells from peripheral blood (PBMC) and SF (SFMC) were collected from healthy individuals and AS. Flow cytometry was used to analyse cells producing inflammatory cytokines. CCL20 levels were determined using ELISA. The impact of CCL20 on Th17 cell migration was verified using a Trans-well migration assay. The in vivo efficacy of CCL20 inhibition was evaluated using an SKG mouse model. RESULTS: The presence of Th17 cells and CCL20 expressing cells was higher in SFMCs from AS patients compared with their PBMCs. The CCL20 level in AS SF was significantly higher than in OA patients. The percentage of Th17 cells in PBMCs from AS patients increased when exposed to CCL20, whereas the percentage of Th17 cells in SFMCs from AS patients decreased when treated with CCL20 inhibitor. The migration of Th17 cells was found to be influenced by CCL20, and this effect was counteracted by the CCL20 inhibitor. In the SKG mouse model, the use of CCL20 inhibitor significantly reduced joint inflammation. CONCLUSION: This research validates the critical role of CCL20 in AS and suggests that targeting CCL20 inhibition could serve as a novel therapeutic approach for AS treatment.

Simultaneous analysis of acylcarnitines using MALDI-TOF mass spectrometry based on a parylene matrix chip.

Short and medium chain acylcarnitines have been used for the diagnosis of various fatty acid oxidation and organic acid disorders. This report presents a multiplex and quantitative analysis of acylcarnitines using MALDI-TOF MS based on a parylene matrix chip. The parylene matrix chip was fabricated by the deposition of a nanoporous film of parylene on an organic matrix array, which reduced the number of mass peaks from the organic matrix in the low m/z range. Quantitative analysis was possible using the parylene matrix chip because of the formation of nano-sized sample crystals on the nanoporous parylene film. Seven acylcarnitines were quantitatively analyzed using the chip; the method detection range included the cut-off values for metabolic disorders. The seven acylcarnitines of different concentrations were simultaneously detected using the parylene matrix chip and the interference from the mixed carnitines was estimated. Real L-carnitine (C0) samples were analyzed using serial dilution, and the recoveries were calculated by comparisons with a standard curve.

Photothermal Structural Dynamics of Au Nanofurnace for In Situ Enhancement in Desorption and Ionization.

Fundamental properties of nanostructured substrates govern the performance of laser desorption/ionization mass spectrometry (LDI-MS); however, limited studies have elucidated the desorption/ionization mechanism based on the physicochemical properties of substrates. Herein, the enhancement in desorption/ionization is investigated using a hybrid matrix of Au nanoisland-functionalized ZnO nanotubes (AuNI-ZNTs). The underlying origin is explored in terms of the photo-electronic and -thermal properties of the matrix. This is the first study to report the effect of laser-induced surface restructuring/melting phenomenon on the LDI-MS performance. AuNI plays a central role as a photothermal nanofurnace, which facilitates the internal energy transfer from the AuNI to the adsorbed analytes by reconstruction in the structurally dynamic AuNI and therefore favors the desorption process. Moreover, piezoelectricity is driven in situ in the AuNI-ZNT hybrid, which modulates the overall band structure and thereby promotes the ionization process. Ultimately, high LDI-MS performance is demonstrated by analyzing small metabolites of fatty acids and monosaccharides, which are challenged to be detected in conventional LDI-MS. This study emphasizing the understanding of matrix properties can provide insights into the design and development of a novel nanomaterial as an efficient LDI matrix. Furthermore, the developed hybrid matrix can overcome the major hurdles existing in conventional LDI-MS.

STAT3 phosphorylation inhibition for treating inflammation and new bone formation in ankylosing spondylitis.

OBJECTIVE: AS is a rheumatic disease characterized by chronic inflammation and bony ankylosis. This study was to evaluate whether a signal transducer and activator of transcription 3 phosphorylation inhibitor (stat3-p Inh) could treat both chronic inflammation and bone formation in AS. METHODS: Primary AS osteoprogenitor cells and spinal entheseal cells were examined for osteogenic differentiation. SF mononuclear cells (SFMCs) and lamina propria mononuclear cells (LPMCs) were obtained from AS patients. Inflammatory cytokine-producing cells were analysed using flow cytometry and ELISA. Female SKG mice were treated with stat3-p Inh, IL-17A blocker or vehicle. Inflammation and new bone formation were evaluated using immunohistochemistry, PET and micro-CT. RESULTS: In the SKG mouse model, stat3-p Inh significantly suppressed arthritis, enthesitis, spondylitis and ileitis. In experiments culturing SFMCs and LPMCs, the frequencies of IFN-γ-, IL-17A- and TNF-α-producing cells were significantly decreased after stat3-p Inh treatment. When comparing current treatments for AS, stat3-p Inh showed a comparable suppression effect on osteogenesis to Janus kinase inhibitor or IL-17A blocker in AS-osteoprogenitor cells. Stat3-p Inh suppressed differentiation and mineralization of AS-osteoprogenitor cells and entheseal cells toward osteoblasts. Micro-CT analysis of hind paws revealed less new bone formation in stat3-p Inh-treated mice than vehicle-treated mice (P = 0.005). Hind paw and spinal new bone formation were similar between stat3-p Inh- and anti-IL-17A-treated SKG mice (P = 0.874 and P = 0.117, respectively). CONCLUSION: Stat-3p inhibition is a promising treatment for both inflammation and new bone formation in AS.

Intralymphatic Administration of Metagonimus yokogawai-Extracted Protein Attenuates Experimental Murine Allergic Rhinitis Model.

OBJECTIVES: This study aimed to evaluate potential therapeutic effect of Metagonimus yokogawai on the OVA-induced allergic rhinitis model. METHODS: OVA-sensitized mice were used to assess potential therapeutic effect of the extract protein of M. yokogawai (My-TP). My-TP was administrated via the intralymphatic route to cervical lymph nodes. The frequencies of sneezing or nasal rubbing were recorded. Histopathologic evaluation was performed for eosinophil infiltrations in the tissues of the nasal mucosa and skin. The mRNA relative expressions of the cytokine profiles including Th1, Th2, Th17, and Treg subsets in the nasal mucosa, cervical lymph nodes, and spleen were analyzed by quantitative real-time reverse-transcriptase polymerase chain reaction. The potential underlying mechanism was investigated by examining cytokine profiles including IL-4 and Treg subsets from lymphocytes of the spleen by flow cytometry. RESULTS: Intralymphatic injection of My-TP reduced allergic symptoms and eosinophil infiltration in the nasal mucosa. My-TP-treated group showed markedly decreased levels of OVA-specific IgE and WBC counts in nasal lavage. My-TP-treated group showed the decreased expression levels of IL-4, while those of IL-10 were increased in both the nasal mucosa. The levels of IFN-γ and IL-17 were also decreased in the nasal mucosa and cervical lymph nodes. The immunological mechanism may involve the downregulation of Th2 response and upregulation of Tregs in the nasal mucosa and cervical lymph nodes. CONCLUSIONS: Our results provide the first evidence of potential therapeutic effect of M. yokogawai in OVA-sensitized allergic rhinitis mice, suggesting that a Treg/Th2 reorganization may play a role in clinical course of allergic rhinitis.

Molecular Detection and Subtyping of Human Blastocystis and the Clinical Implications: Comparisons between Diarrheal and Non-diarrheal Groups in Korean Populations.

Blastocystis has recently been recognized as the most common eukaryotic microbe of the human gut. We investigated the prevalence of Blastocystis and their subtypes in diarrheal and non-diarrheal groups and the associated clinical parameters. A total of 324 stool samples were obtained from 196 diarrheal and 128 non-diarrheal subjects. Blastocystis subtypes were determined by sequencing the small subunit ribosomal DNA (SSU rRNA) gene. Demographic, clinical and laboratory data were collected and analyzed by diarrhea and Blastocystis status. The overall rate of Blastocystis positivity was 9.0% (29/324) but was significantly higher in the non-diarrheal group (18.0% vs. 3.1%, P<0.0001). Of the 6 Blastocystis-positive diarrheal patients, 3 (50.0%), none (0.0%), 2 (33.3%), and 1 (16.7%) were infected with subtypes ST1, ST2, ST3, and multiple subtypes, respectively. Of the 23 Blastocystis-positive non-diarrheal patients, 4 (17.4%), 1 (4.3%), and 18 (78.3%) were infected with subtypes ST1, ST2, and ST3, respectively. Blastocystis was less common in the diarrheal than the non-diarrheal group (odds ratio, 0.144; 95% confidence interval, 0.057-0.365, P<0.001). Of the 3 subtypes, ST3 was more frequently observed in the non-diarrheal than diarrheal group (78.3% vs. 33.3%, P=0.0341). Collectively, Blastocystis was found in both the diarrheal and non-diarrheal groups and ST3 was the most common subtype in Korea.

Morphological and Molecular Characteristics of Clinostomid Metacercariae from Korea and Myanmar.

Morphological and molecular characterization of clinostomid metacercariae (CMc) was performed with the specimens collected in fish from Korea and Myanmar. Total 6 batches of clinostomid specimens by the fish species and geographical localities, 5 Korean and 1 Myanmar isolates, were analyzed with morphological (light microscopy and SEM) and molecular methods (the cytochrome c oxidase 1 gene and internal transcribed spacer 1/5.8S rRNA sequence). There were some morphological variations among CMc specimens from Korea. However, some morphometrics, i.e., the size of worm body and each organ, ratio of body length to body width, and morphology of cecal lumens, were considerably different between the specimens from Korea and Myanmar. The surface ultrastructures were somewhat different between the specimens from Korea and Myanmar. The CO1 sequences of 5 Korean specimens ranging 728-736 bp showed 99.6-100% identity with Clinostomum complanatum (GenBank no. KM923964). They also showed 99.9-100% identity with C. complanatum (FJ609420) in the ITS1 sequences ranging 692-698 bp. Meanwhile, the ITS1 sequences of Myanmar specimen showed 99.9% identity with Euclinostomum heterostomum (KY312847). Five sequences from Korean specimens clustered with the C. complanatum genes, but not clustered with Myanmar specimens. Conclusively, it was confirmed that CMc from Korea were morphologically and molecularly identical with C. complanatum and those from Myanmar were E. heterostomum.

Nanostructured TiO2 Materials for Analysis of Gout-Related Crystals Using Laser Desorption/Ionization Time-of-Flight (LDI-ToF) Mass Spectrometry.

Crystals of monosodium urate monohydrate (MSU) and calcium pyrophosphate dihydrate (CPPD) are known to induce arthropathic diseases called gout and pseudogout, respectively. These crystals are deposited in various joints or tissues, causing severe pain. Correct identification of crystals is crucial for the appropriate treatment of gout and pseudogout, which exhibit very similar symptoms. Herein, a novel approach of laser desorption/ionization time-of-flight (LDI-ToF) mass spectrometry (MS) was introduced to analyze MSU and CPPD crystals with three different types of nanostructured TiO2 materials including TiO2 nanoparticles (P25), TiO2 nanowires synthesized from wet-corrosion method, and the mixture of P25 and TiO2 nanowires (P25/TiO2 nanowires) as inorganic solid matrices. Furthermore, the feasibility of LDI-ToF MS based on these TiO2 nanostructures for the analysis of the two arthropathy-related crystals was tested using spiked samples in synovial fluid at known crystal concentrations. The mass analysis results of MSU and CPPD crystals demonstrated that (1) the electrostatic interaction between analytes and solid matrices was key for the analyte ionization and (2) LDI-ToF MS with nanostructured TiO2 materials has the potential to be a practical approach for the diagnosis of gout and pseudogout.

MALDI-TOF Mass Spectrometry Based on Parylene-Matrix Chip for the Analysis of Lysophosphatidylcholine in Sepsis Patient Sera.

In this work, medical diagnosis of sepsis was conducted via quantitative analysis of lysophosphatidylcholine 16:0 (LPC 16:0) by using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry based on a parylene-matrix chip. In the first step, specific mass peaks for the diagnosis of sepsis were searched by comparing MALDI-TOF mass spectra of sepsis patient sera with healthy controls and pneumonia patient sera. Two mass peaks at m/z = 496.3 and 518.3 were chosen as those that are specifically different for sepsis sera to compare with healthy controls and pneumonia patient sera. These mass peaks were identified to be protonated and sodium adducts of LPC 16:0 by using tandem mass spectra (MS2 and MS3) of purely synthesized LPC 16:0 and extracted LPC 16:0 from a healthy control and a sepsis patient. In the next step, a standard curve for LPC 16:0 for the quantitative analysis of LPC 16:0 with MALDI-TOF MS based on the parylene-matrix chip was prepared, and the statistical correlation to the LC-MS analysis results was demonstrated by using the Bland-Altman test and Passing-Bablok regression. Finally, MALDI-TOF MS based on the parylene-matrix chip was used for the quantification of LPC 16:0 with sera from patients with severe sepsis and septic shock (n = 143), pneumonia patients (n = 12), and healthy sera (n = 31). The sensitivity and the selectivity of medical diagnosis of sepsis was estimated to be 97.9% and 95.5% by using MALDI-TOF MS based on the parylene-matrix chip, respectively.

Gold-nanoparticle-coated magnetic beads for concentration and ionization of analytes for laser desorption/ionization mass spectrometry.

RATIONALE: Magnetic particles coated with gold nanoparticles (Au NPs) (Au-MAGs) were developed and used (1) for sample concentration and (2) as a solid matrix for laser adsorption/desorption mass spectrometry (LDI-MS). METHODS: The Au-MAGs were prepared by (1) coating polystyrene on iron oxide NPs (PS-MNPs), (2) coating poly-l-lysine on the PS-MNPs (PLL-coated PS-MNPs), and (3) coating negatively charged Au NPs on the PLL-coated PS-MNPs (Au-MAGs). RESULTS: The Au-MAGs were used to concentrate the target analyte by means of electrostatic interactions between the positively charged GHP9 and the negatively charged Au-MAGs as well as selective interactions (such as gold-sulfur interactions) between glutathione (GSH) and Au-MAGs. Then, the concentrated analyte could be ionized for LDI-MS. CONCLUSIONS: The Au-MAGs were demonstrated (1) to concentrate the target analyte in a sample solution, tested by electrostatic interactions and selective interactions between gold and sulfur and (2) to ionize the concentrated analyte for LDI-MS.

Four Taeniasis saginata Cases Diagnosed at a University Hospital in Korea.

In recent years, the taeniasis has been rarely reported in the Republic of Korea (Korea). But in this study, we intend to report 4 taeniasis cases caused by Taenia saginata during a 5-month period (February to June 2018) at a unversity hospital in Gwangju, Korea. Worm samples (proglottids) discharged from all cases were identified by phenotypic and molecular diagnostics. Mitochondrial cytochrome c oxidase subunit I sequences showed 99.4-99.9% identity with T. saginata but, differed by 4% from T. asiatica and by 7% from T. multiceps, respectively. We found that tapeworms in 2 cases (Cases 2 and 3) yielded exactly the same sequences between them, which differed from those in Cases 1 and 4, suggesting intra-species variation in tapeworms. These taeniasis cases by T. saginata infection in this study, which occurred within a limited time period and region, suggest the possibility of a mini-outbreak. This study highlights the need for further epidemiological investigation of potentially overlooked cases of T. saginata infection in Korea.

Crosscultural Adaptation and Validation of the Korean Version of the New Knee Society Knee Scoring System.

BACKGROUND: The 2011 Knee Society Score© (2011 KS Score©) is used to characterize the expectations, symptoms, physical activity, and satisfaction of patients who undergo TKA and is widely used to assess the outcome of TKA. However, it has not been adapted or validated for use in Korea. QUESTIONS/PURPOSES: We developed a Korean version of the 2011 KS Score and evaluated the (1) test-retest reliability, (2) convergent validity, and (3) responsiveness of the Korean version. METHODS: The Korean version of the 2011 KS Score was derived by using a well-established translational procedure based on international guidelines, which include translation, synthesis, back-translation, expert committee review, pretesting, and submission for appraisal. A total of 123 patients with knee osteoarthritis who were scheduled to undergo TKA were recruited for the study. Ninety percent of the patients (111 of 123) were women, which is an exact representation of the Korean population having TKAs. To evaluate reliability, the patients were evaluated twice during a 4-week interval using the questionnaire. Reliability was assessed by using intraclass correlation coefficients (ICCs) and internal consistency by using Cronbach's alpha to determine the validity of the Korean version of the 2011 KS Score. The patients were evaluated by using the validated Korean versions of the WOMAC and SF-36 questionnaires. Spearman's correlation coefficient was used for validation. Responsiveness was determined by calculating the standardized response mean from the preoperative and postoperative test scores in the Korean version of the 2011 KS Score. To address the gender disparity in our study we identified 53 males who underwent TKA for osteoarthritis after completion of this study and generated age-matched controlled groups to evaluate construct validity and responsiveness in Korean males. RESULTS: The reliability proved good to excellent with an ICC between 0.69 and 0.85, depending on the clinical properties tested, which included the following: symptoms, satisfaction, expectation, and total functional activity consisting of functional activity, standard activity, advanced activity, and discretionary activity. All subscales showed good to excellent internal consistency indicated by Chronbach's alpha (range, 0.83-0.92). For validity, three of the four domains (the exception was expectation) of the 2011 KS Score, correlated either strongly or moderately with the Korean WOMAC score (r ≥ 0.35). When compared with the SF-36, the satisfaction domain showed a weak positive correlation with all the subscales of the SF-36 except general health (r < 0.35). The activity domain showed a strong positive correlation with physical function (r = 0.62) and physical component summary (r = 0.52), moderate with physical role (r = 0.46), and weak with bodily pain (r = 0.26) and social function (r = 0.31). The symptom domain also exhibited a similar moderate positive correlation with physical function (r = 0.41) and weak positive correlation with bodily pain, social function, and physical component summary (r = 0.22, 0.20, and 0.26, respectively). For responsiveness, all the domains of Korean version of the 2011 KS Score, except for expectation, showed large changes (> 0.8), calculated as standardized response mean. The total amount of the Korean version of the 2011 KS Score (2.03, p < 0.001) showed higher responsiveness when compared with the WOMAC total (1.88, p < 0.001) and SF-36 physical and mental component summaries (1.14, p < 0.001; and 0.68, p < 0.001, respectively). CONCLUSIONS: The Korean version of the 2011 KS Score was successfully developed using a process of crosscultural adaptation for the Korean-speaking population who had undergone TKA for osteoarthritis of the knee. The Korean version of the 2011 KS Score was shown to be a reliable, valid, and responsive tool and can be used to assess functional outcomes and expectations of Korean patients who undergo TKA. The demographic features of TKA in the Korean population should be taken into account with additional studies recommended to further investigate these psychometric properties in Korean men. LEVEL OF EVIDENCE: Level II, diagnostic study.

Mucosal-associated invariant T cell deficiency in systemic lupus erythematosus.

Mucosal-associated invariant T (MAIT) cells contribute to protection against certain microorganism infections and play an important role in mucosal immunity. However, the role of MAIT cells remains enigmatic in autoimmune diseases. In this study, we examined the level and function of MAIT cells in patients with rheumatic diseases. MAIT cell, cytokine, and programmed death-1 (PD-1) levels were measured by flow cytometry. Circulating MAIT cell levels were significantly reduced in systemic lupus erythematosus (SLE) and rheumatoid arthritis patients. In particular, this MAIT cell deficiency was more prominent in CD8(+) and double-negative T cell subsets, and significantly correlated with disease activity, such as SLE disease activity index and 28-joint disease activity score. Interestingly, MAIT cell frequency was significantly correlated with NKT cell frequency in SLE patients. IFN-γ production in MAIT cells was impaired in SLE patients, which was due to an intrinsic defect in the Ca(2+)/calcineurin/NFAT1 signaling pathway. In SLE patients, MAIT cells were poorly activated by α-galactosylceramide-stimulated NKT cells, thereby showing the dysfunction between MAIT cells and NKT cells. Notably, an elevated expression of PD-1 in MAIT cells and NKT cells was associated with SLE. In rheumatoid arthritis patients, MAIT cell levels were significantly higher in synovial fluid than in peripheral blood. Our study primarily demonstrates that MAIT cells are numerically and functionally deficient in SLE. In addition, we report a novel finding that this MAIT cell deficiency is associated with NKT cell deficiency and elevated PD-1 expression. These abnormalities possibly contribute to dysregulated mucosal immunity in SLE.

Mucosal-Associated Invariant T Cell Deficiency in Chronic Obstructive Pulmonary Disease.

Mucosal-associated invariant T (MAIT) cells have been reported to play an important role in mucosal immunity. However, little is known about the roles of MAIT cells in chronic obstructive pulmonary disease (COPD). The aims of this study were to examine the levels of circulating MAIT cells and their subsets in COPD patients and to investigate the potential relationship between clinical parameters and MAIT cell levels. Forty-five COPD patients and 57 healthy control subjects were enrolled in the study. Circulating MAIT cells and their subset levels in the peripheral blood were measured by flow cytometry. Disease grades were classified according to the GOLD criteria for the assessment of severity of COPD. Circulating MAIT cell levels were found to be significantly reduced in COPD patients. In particular, this MAIT cell deficiency was more prominent in CD8+ and double-negative T cell subsets. Interestingly, elevated serum C-reactive protein level and reduced FEV1/FVC ratio were associated with MAIT cell deficiency in COPD patients. Furthermore, the circulating MAIT levels were found to be significantly lower in patients with moderate to severe COPD than in patients with mild COPD. Our data shows that MAIT cells are numerically deficient in the peripheral blood of patients with COPD. In addition, this MAIT cell deficiency was found to reflect inflammatory activity and disease severity. These findings provide important information for monitoring the changes in MAIT cell levels and for predicting the prognosis during the disease course.

Analysis of predictors influencing indeterminate whole-blood interferon-gamma release assay results in patients with rheumatic diseases.

Triggers of indeterminate results from interferon-gamma release assays (IGRA) in patients with rheumatic diseases are still elusive. The aim of the present study was to describe predictors of indeterminate results from IGRA in the field of rheumatology. This cross-sectional study was retrospectively performed by using a database of patients with a request for QuantiFERON-TB Gold-In Tube test (QFT-GIT) for screening of latent tuberculosis infection. The study cohort included 631 patients with rheumatic diseases. All variables influencing indeterminate QFT-GIT results were investigated by logistic regression analysis. The overall frequency of indeterminate IGRA results was 6.8 % (43/631). Those with indeterminate results were more likely to be aged ≥70 years, female, visitors in winter, suffering from systemic lupus erythematosus (SLE), and using sulfasalazine or a tumor necrosis factor (TNF)-α inhibitor. In addition, a longer incubation time of >6 h increased the odds ratio of indeterminate IGRA results. In contrast, the automated ELISA processor, ankylosing spondylitis, and the use of a non-steroidal anti-inflammatory drug decreased the likelihood of indeterminate IGRA results. Lymphopenia, thrombocytopenia, anemia, and hypoalbuminemia were significantly associated with indeterminate IGRA results. Multivariate analysis revealed that SLE, use of sulfasalazine or a TNF-α inhibitor, and a manual ELISA system were significantly independent predictors of indeterminate IGRA results. The proportion of indeterminate results in patients with rheumatic diseases is not infrequent. Careful attention to the pre-analytical conditions should minimize the indeterminate results. Automation of the ELISA process seems to be a promising solution to decrease the rate of indeterminate response.

The effectiveness of tumor necrosis factor-α blocker therapy in patients with axial spondyloarthritis who failed conventional treatment: a comparative study focused on improvement in ASAS Health Index.

Objective: The purpose of this study is to evaluate the impact of tumor necrosis factor (TNF)-α blocker therapy on the Assessment of SpondyloArthritis international Society Health Index (ASAS-HI) among patients who have failed conventional nonsteroidal anti-inflammatory drugs. Methods: A comparative study was conducted involving axial spondyloarthritis (axSpA) patients treated with either TNF-α blocker or conventional therapy. Patient data, including demographics, disease characteristics, and ASAS-HI scores, were collected before and after treatment. Statistical analysis was performed to compare changes in ASAS-HI scores between the TNF-α blocker and the conventional therapy group. Results: The study population consisted of patients with axSpA, with a mean age of 38.3 years in conventional treatment group and 29.3 years in TNF-α blocker group. Most variables, including C-reactive protein levels, other comorbidities, and disease assessment scores showed no significant difference between groups. Longitudinal analysis within each treatment group from Week 0 to 12 showed no significant change in the conventional treatment group, whereas the TNF-α blocker group experienced a significant reduction in ASAS-HI scores, demonstrating the effectiveness of the treatment. The TNF-α blocker group exhibited a significantly greater improvement in ASAS-HI scores compared to the conventional therapy group. The Bath Ankylosing Spondylitis Functional Index and the Bath Ankylosing Spondylitis Disease Activity Index demonstrated strong positive correlations with ASAS-HI scores, indicating higher disease activity and functional limitation are associated with worse health outcomes in patients. Conclusion: The research demonstrates that ASAS-HI scores significantly improve with TNF-α blocker therapy in axSpA patients, underscoring ASAS-HI's effectiveness as a tool for evaluating drug responses.

Lower respiratory tract microbiota in patients with clinically suspected nontuberculous mycobacterial pulmonary disease according to the presence of gastroesophageal reflux.

Although gastroesophageal reflux has been recognized as one of the risk factors of nontuberculous mycobacterial pulmonary disease (NTM-PD) progression, the effect of reflux on the lower respiratory tract microbiota has not been studied in detail. We investigated the composition of the lower respiratory tract microbiota in patients with clinically suspected NTM-PD, comparing them based on the presence of reflux. Forty-seven patients suspected of having NTM-PD were enrolled and assigned according to presence of reflux (n = 22) and non- reflux (n = 25). We performed a pepsin ELISA assay to identify the presence of reflux and 16S ribosomal RNA gene amplicon sequencing to evaluate the microbiota in bronchoalveolar lavage fluid. There were no significant differences in the diversity or composition of the lower respiratory microbiota between the NTM-PD and non-NTM-PD groups. Bacterial richness was observed in the non-reflux group than in the reflux group [P = 0.03] and a cluster in the reflux group was observed. The reflux group showed a predominance for Pseudomonas aeruginosa or Staphylococcus aureus among the NTM-PD group and for P. aeruginosa, Haemophilus influenzae, Klebsiella pneumoniae, or Eikenella species among the non-NTM-PD group. The non-reflux groups presented diverse patterns. A linear discriminant analysis and volcano plot demonstrated that P. aeruginosa, H. haemolyticus, Selenomonas artemidis, and Dolosigranulum pigrum were specifically associated with the NTM-PD reflux group, while P. aeruginosa was specifically associated with the non-NTM-PD reflux group. These observations confirm that the lower respiratory microbiota is consistently altered by reflux but not in NTM-PD.

Exosome-mediated delivery of super-repressor IκBα alleviates inflammation and joint damages in rheumatoid arthritis.

BACKGROUND: This study aims to investigate the potential anti-inflammatory effects of exosomes engineered to carry super-repressor IκB (Exo-srIκB), an exosome-based NF-κB inhibitor, in the context of RA. METHODS: Peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) were collected from patients diagnosed with RA and treated with Exo-srIκB to test the therapeutic potential. Flow cytometry analysis was performed to assess the production of inflammatory cytokines (IL-17A and GM-CSF) by the cells. ELISA was utilized to measure the levels of TNF-α, IL-17A, IL-6, and GM-CSF. Arthritis was induced in SKG mice by intraperitoneal injection of curdlan. DBA/1 J mice were used in collagen-induced arthritis (CIA) experiments. After the development of arthritis, mice were injected with either Exo-Naïve (control exosome) or Exo-srIκB. Arthritis scores were recorded biweekly, and histological observations of the ankle joint were conducted using H&E and safranin-O staining. Additionally, bone erosion was evaluated using micro-CT imaging. RESULTS: In the ex vivo study involving human PBMCs and SFMCs, treatment with Exo-srIκB demonstrated a notable reduction in inflammatory cytokines. Furthermore, in both the SKG and CIA models, Exo-srIκB treatment exhibited significant reductions in inflammation, cartilage destruction, and bone erosion within the joint tissues when compared to the Exo-Naive control group. Additionally, the radiographic score assessed through microCT showed a significant decrease compared to the Exo-Naive control group. CONCLUSION: Overall, these findings suggest that Exo-srIκB possesses anti-inflammatory properties in human RA cells and animal models, making it a promising therapeutic candidate for the treatment of RA.