ABSTRACT
This case report describes rare concomitant allergic fungal rhinosinusitis (AFRS) and chronic granulomatous invasive fungal sinusitis (CGIFS) in a 34-year-old woman with acute lymphoblastic leukemia and graft-versus-host disease (GVHD) post bone marrow transplantation. Initially presenting with rhinorrhea and nasal obstruction, the patient was diagnosed with AFRS in the right maxillary sinus, followed by a postoperative course of CGIFS in the left nasal cavity, showcasing the unique occurrence. She was not immunocompromised during diagnosis. CGIFS may have occurred because of surgery; however, voriconazole led to significant improvement. This case highlights noninvasive and invasive fungal infections in patients with chronic rhinosinusitis and a history of GVHD and underscores the complexity of diagnosing and managing such cases.
ABSTRACT
Non-thermal atmospheric-pressure plasma (NTAPP) has been widely studied for clinical applications, e.g., disinfection, wound healing, cancer therapy, hemostasis, and bone regeneration. It is being revealed that the physical and chemical actions of plasma have enabled these clinical applications. Based on our previous report regarding plasma-stimulated bone regeneration, this study focused on Achilles tendon repair by NTAPP. This is the first study to reveal that exposure to NTAPP can accelerate Achilles tendon repair using a well-established Achilles tendon injury rat model. Histological evaluation using the Stoll's and histological scores showed a significant improvement at 2 and 4 weeks, with type I collagen content being substantial at the early time point of 2 weeks post-surgery. Notably, the replacement of type III collagen with type I collagen occurred more frequently in the plasma-treated groups at the early stage of repair. Tensile strength test results showed that the maximum breaking strength in the plasma-treated group at two weeks was significantly higher than that in the untreated group. Overall, our results indicate that a single event of NTAPP treatment during the surgery can contribute to an early recovery of an injured tendon.
Subject(s)
Achilles Tendon , Plasma Gases , Tendon Injuries , Wound Healing , Animals , Achilles Tendon/injuries , Rats , Plasma Gases/pharmacology , Plasma Gases/therapeutic use , Wound Healing/drug effects , Tendon Injuries/therapy , Male , Helium/pharmacology , Rats, Sprague-Dawley , Collagen Type I/metabolism , Tensile Strength , Atmospheric Pressure , Collagen Type III/metabolismABSTRACT
Purpose: The purpose of this study was to investigate the incidence of anomalies in patients who underwent endoscopic carpal tunnel release and their relationship with clinical outcomes. Methods: This retrospective study included 65 hands of 57 patients (8 men and 49 women; mean age, 64.9 years) who underwent endoscopic carpal tunnel release for carpal tunnel syndrome at our hospital between March 2016 and April 2022. The patients were diagnosed with carpal tunnel syndrome based on clinical observations and electrophysiological studies. On T2-weighted magnetic resonance axial images, the height of the hook of the hamate was measured from the bottom to the tip of the hook, and the total height of the hamate was measured from the dorsal surface of the hamate to the tip of the hook. A hook-to-height ratio of less than 0.34 was defined as hypoplastic, and its incidence was investigated. In addition, electrodiagnostic testing of sensory and motor nerve conduction of the median nerve and patient-reported outcome measurements, including Quick Disabilities of the Arm, Shoulder and Hand score, Boston carpal tunnel questionnaire, and visual analog scale score, were investigated at 6 months after surgery. Adverse events were collected from patient records. Results: The mean hook-to-height ratio was 0.40. Hypoplasia with a ratio ≤0.34 was observed in seven hands (10.8%), and adverse events were observed only in the two cases that had a hypoplastic hook of the hamate (3.07%). The patient-reported outcome measurements and the result of electrodiagnostic testing at 6 months after surgery did not correlate with the height of the hook of the hamate. Conclusions: The incidence of a hypoplastic hook of the hamate is common in patients with carpal tunnel syndrome, and preoperative evaluation of the morphology of the hooks and indications for endoscopic carpal tunnel release in cases of hypoplastic hooks may help predict adverse events. Type of study/level of evidence: Therapeutic â £.
ABSTRACT
Non-thermal atmospheric-pressure plasma (NTAPP) is attracting widespread interest for use in medical applications. The tissue repair capacity of NTAPP has been reported in various fields; however, little is known about its effect on fracture healing. Non-union or delayed union after a fracture is a clinical challenge. In this study, we aimed to investigate how NTAPP irradiation promotes fracture healing in a non-union fracture model and its underlying mechanism, in vitro and in vivo. For the in vivo study, we created normal and non-union fracture models in LEW/SsNSlc rats to investigate the effects of NTAPP. To create a fracture, a transverse osteotomy was performed in the middle of the femoral shaft. To induce the non-union fracture model, the periosteum surrounding the fracture site was cauterized after a normal fracture model was created. The normal fracture model showed no significant difference in bone healing between the control and NTAPP-treated groups. The non-union fracture model demonstrated that the NTAPP-treated group showed consistent improvement in fracture healing. Histological and biomechanical assessments confirmed the fracture healing. The in vitro study using pre-osteoblastic MC3T3-E1 cells demonstrated that NTAPP irradiation under specific conditions did not reduce cell proliferation but did enhance osteoblastic differentiation. Overall, these results suggest that NTAPP is a novel approach to the treatment of bone fractures.
Subject(s)
Femoral Fractures , Fractures, Bone , Plasma Gases , Rats , Animals , Fracture Healing , Plasma Gases/pharmacology , Plasma Gases/therapeutic use , Cell Differentiation , Cell Proliferation , Femoral Fractures/surgeryABSTRACT
Validation of biomarker assays is crucial for effective drug development and clinical applications. Interlaboratory reproducibility is vital for reliable comparison and combination of data from different centers. This review summarizes interlaboratory studies of quantitative LC-MS-based biomarker assays using reference standards for calibration curves. The following points are discussed: trends in reports, reference and internal standards, evaluation of analytical validation parameters, study sample analysis and normalization of biomarker assay data. Full evaluation of these parameters in interlaboratory studies is limited, necessitating further research. Some reports suggest methods to address variations in biomarker assay data among laboratories, facilitating organized studies and data combination. Method validation across laboratories is crucial for reducing interlaboratory differences and reflecting target biomarker responses.
Subject(s)
Liquid Chromatography-Mass Spectrometry , Tandem Mass Spectrometry , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Reproducibility of Results , Reference StandardsABSTRACT
Lipid mediators have been suggested to play important roles in the pathogenesis of rheumatoid arthritis (RA). Lipidomics has recently allowed for the comprehensive analysis of lipids and has revealed the potential of lipids as biomarkers for the early diagnosis of RA and prediction of therapeutic responses. However, the relationship between disease activity and the lipid profile in RA remains unclear. In the present study, we performed a plasma lipidomic analysis of 278 patients with RA during treatment and examined relationships with disease activity using the Disease Activity Score in 28 joints (DAS28)-erythrocyte sedimentation rate (ESR). In all patients, five lipids positively correlated and seven lipids negatively correlated with DAS28-ESR. Stearic acid [FA(18:0)] (r = -0.45) and palmitic acid [FA(16:0)] (r = -0.38) showed strong negative correlations. After adjustments for age, body mass index (BMI), and medications, stearic acid, palmitic acid, bilirubin, and lysophosphatidylcholines negatively correlated with disease activity. Stearic acid inhibited osteoclast differentiation from peripheral blood monocytes in in vitro experiments, suggesting its contribution to RA disease activity by affecting bone metabolism. These results indicate that the lipid profile correlates with the disease activity of RA and also that some lipids may be involved in the pathogenesis of RA.
ABSTRACT
BACKGROUND: Transoral videolaryngoscopic surgery (TOVS) is widely used in Japan, and conventional two-dimensional (2D) endoscopic methods have been established. Three-dimensional (3D) endoscopic surgery offers superior distance perception because it provides stereoscopic views. Recently, we have developed 3D endoscopy for TOVS (3D TOVS). METHODS: This study included 46 patients with pharyngeal cancer who underwent 3D TOVS. The perioperative complications and survival curves were retrospectively analyzed. RESULTS: One patient with oropharyngeal cancer who underwent neck dissection and transoral resection simultaneously experienced postoperative hemorrhage of the neck. Another patient with oropharyngeal cancer underwent hemostasis for postoperative pharyngeal hemorrhage. There was one case of aspiration pneumonia. One patient developed cervical lymph node recurrence; however, there was no local recurrence or primary mortality. The 2-year overall survival, disease-specific survival, local control rates, locoregional control rate, and invasive disease-free survival were 90.9%, 100%, 100%, 97.4%, and 79.9%, respectively. CONCLUSIONS: Three-dimensional endoscopy can be safely applied to TOVS.
Subject(s)
Laryngoscopy , Video-Assisted Surgery , Humans , Male , Female , Middle Aged , Aged , Retrospective Studies , Laryngoscopy/methods , Video-Assisted Surgery/methods , Imaging, Three-Dimensional , Adult , Aged, 80 and over , Treatment Outcome , Natural Orifice Endoscopic Surgery/methods , Natural Orifice Endoscopic Surgery/adverse effects , Pharyngeal Neoplasms/surgery , Pharyngeal Neoplasms/mortality , Pharyngeal Neoplasms/pathology , Japan , Neck Dissection , Disease-Free SurvivalABSTRACT
BACKGROUND: Drug-induced interstitial lung disease (DILD) is a lung injury caused by various types of drugs and is a serious problem in both clinical practice and drug development. Clinical management of the condition would be improved if there were DILD-specific biomarkers available; this study aimed to meet that need. METHODS: Biomarker candidates were identified by non-targeted metabolomics focusing on hydrophilic molecules, and further validated by targeted approaches using the serum of acute DILD patients, DILD recovery patients, DILD-tolerant patients, patients with other related lung diseases, and healthy controls. RESULTS: Serum levels of kynurenine and quinolinic acid (and kynurenine/tryptophan ratio) were elevated significantly and specifically in acute DILD patients. The diagnostic potentials of these biomarkers were superior to those of conventional lung injury biomarkers, Krebs von den Lungen-6 and surfactant protein-D, in discriminating between acute DILD patients and patients with other lung diseases, including idiopathic interstitial pneumonia and lung diseases associated with connective tissue diseases. In addition to identifying and evaluating the biomarkers, our data showed that kynurenine/tryptophan ratios (an indicator of kynurenine pathway activation) were positively correlated with serum C-reactive protein concentrations in patients with DILD, suggesting the potential association between the generation of these biomarkers and inflammation. Our in vitro experiments demonstrated that macrophage differentiation and inflammatory stimulations typified by interferon gamma could activate the kynurenine pathway, resulting in enhanced kynurenine levels in the extracellular space in macrophage-like cell lines or lung endothelial cells. Extracellular quinolinic acid levels were elevated only in macrophage-like cells but not endothelial cells owing to the lower expression levels of metabolic enzymes converting kynurenine to quinolinic acid. These findings provide clues about the molecular mechanisms behind their specific elevation in the serum of acute DILD patients. CONCLUSIONS: The serum concentrations of kynurenine and quinolinic acid as well as kynurenine/tryptophan ratios are promising and specific biomarkers for detecting and monitoring DILD and its recovery, which could facilitate accurate decisions for appropriate clinical management of patients with DILD.
Subject(s)
Lung Diseases, Interstitial , Lung Injury , Humans , Kynurenine/metabolism , Tryptophan/metabolism , Tryptophan/pharmacology , Quinolinic Acid/metabolism , Endothelial Cells/metabolism , Lung Diseases, Interstitial/chemically induced , Lung Diseases, Interstitial/diagnosis , BiomarkersABSTRACT
Recently, animal welfare has been attracting worldwide attention, and implementation of 3Rs (replacement, reduction, and refinement) is prioritized in every way possible in the drug development. Microsampling, in which small amounts of blood are collected, is attracting attention in this context. ICH S3A Q&A focused on microsampling was published in November 2017 to help accelerate the application of microsampling for toxicokinetic assessment. The increased sensitivity of drug measurement apparatuses such as mass spectrometers has made it possible to measure drug concentrations with small amounts of blood samples. In this review, we summarized the reports on toxicological influence of microsampling in rodents (rats and mice) with or without drug administration or recovery period after blood collection and influences that may arise from differences in the blood sampling site or blood sampling volume. We also summarized some perspectives on further implementation of microsampling in toxicology studies. The use of microsampling in regulatory toxicology studies has gradually increased, although at a lower rate than in discovery studies. Since more animals are used in GLP toxicology studies than in discovery studies, the effect of reducing the number of animals by microsampling is expected to be greater in the toxicology studies. This report aims to promote the application of microsampling to nonclinical studies, as it is beneficial for improving animal welfare and can contribute to the 3Rs.
Subject(s)
Blood Specimen Collection , Rodentia , Rats , Mice , Animals , Mass SpectrometryABSTRACT
Isolated injury to the deep motor branch of the ulnar nerve caused by stabbing is sporadic, with only one reported case in the English-language literature. We report one such case treated successfully using nerve grafting. A 33-year-old patient had sustained a stab wound to the right hypothenar eminence and showed a claw hand deformity. Needle electromyography study revealed denervation potentials with no voluntary motor unit action potentials (MUAPs) in the first dorsal interosseous (FDI) muscles. Nerve exploration revealed a neuroma-in-continuity in the intrinsic motor branch of the ulnar nerve. Intraoperative nerve stimulation confirmed the absence of compound muscle action potentials in the FDI. The damaged scarred nerve was resected, and the 15-mm defects were reconstructed with cable autografting. Two years and 5 months after the surgery, voluntary MUAPs were observed in the FDI. The pinch strengths recovered. Laceration of the deep branch of the ulnar nerve caused by stabbing can sometimes remain hidden as the hand sensation remains intact. Pre- and intraoperative electrophysiological examination is essential to assess the severity of the injured nerve and determine an appropriate surgical option. Even nerve grafting can facilitate satisfactory results as target intrinsic muscles are quite close to the repair site.
ABSTRACT
ATSP-7041, a stapled α-helical peptide that inhibits murine double minute-2 (MDM2) and MDMX activities, is a promising modality targeting protein-protein interactions. As peptides of molecular weights over 1000 Da are not usually evaluated, data on the drug-drug interaction (DDI) potential of stapled α-helical peptides remain scarce. Here, we evaluate the interaction of ATSP-7041 with hepatic cytochrome P450s (CYPs; CYP1A2, CYP2C9, CYP2C19, CYP3A4, and CYP2D6) and transporters (organic anion transporting polypeptides (OATPs; OATP1B1 and OATP1B3), P-glycoprotein (P-gp), and breast cancer resistance protein (BCRP)). ATSP-7041 demonstrated negligible metabolism in human liver S9 fraction and a limited inhibition of CYP activities in yeast microsomes or S9 fractions. On the contrary, a substantial uptake by OATPs in HEK 293 cells, a strong inhibition of OATP activities in the cells, and an inhibition of P-gp and BCRP activities in reversed membrane vesicles were observed for ATSP-7041. A recent report describes that ALRN-6924, an ATSP-7041 analog, inhibited OATP activities in vivo; therefore, we focused on the interaction between ATSP-7041 and OATP1B1 to demonstrate that ATSP-7041, as a higher molecular weight stapled peptide, is a substrate and strong inhibitor of OATP1B1 activity. Our findings demonstrated the possibility of transporter-mediated DDI potential by high molecular weight stapled peptides and the necessity of their evaluation for drug development.
Subject(s)
Neoplasm Proteins , Organic Anion Transporters , Humans , Mice , Animals , ATP Binding Cassette Transporter, Subfamily G, Member 2 , HEK293 Cells , Neoplasm Proteins/metabolism , Peptides/pharmacology , Peptides/metabolism , Membrane Transport Proteins/metabolism , Organic Anion Transporters/metabolism , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Cytochrome P-450 Enzyme System/metabolismABSTRACT
Large animal experiments are important for preclinical studies of regenerative stem cell transplantation therapy. Therefore, we investigated the differentiation capacity of pig skeletal muscle-derived stem cells (Sk-MSCs) as an intermediate model between mice and humans for nerve muscle regenerative therapy. Enzymatically extracted cells were obtained from green-fluorescence transgenic micro-mini pigs (GFP-Tg MMP) and sorted as CD34+/45- (Sk-34) and CD34-/45-/29+ (Sk-DN) fractions. The ability to differentiate into skeletal muscle, peripheral nerve, and vascular cell lineages was examined via in vitro cell culture and in vivo cell transplantation into the damaged tibialis anterior muscle and sciatic nerves of nude mice and rats. Protein and mRNA levels were analyzed using RT-PCR, immunohistochemistry, and immunoelectron microscopy. The myogenic potential, which was tested by Pax7 and MyoD expression and the formation of muscle fibers, was higher in Sk-DN cells than in Sk-34 cells but remained weak in the latter. In contrast, the capacity to differentiate into peripheral nerve and vascular cell lineages was significantly stronger in Sk-34 cells. In particular, Sk-DN cells did not engraft to the damaged nerve, whereas Sk-34 cells showed active engraftment and differentiation into perineurial/endoneurial cells, endothelial cells, and vascular smooth muscle cells, similar to the human case, as previously reported. Therefore, we concluded that Sk-34 and Sk-DN cells in pigs are closer to those in humans than to those in mice.
Subject(s)
Endothelial Cells , Muscle Fibers, Skeletal , Mice , Humans , Rats , Animals , Swine , Mice, Nude , Swine, Miniature , Muscle Fibers, Skeletal/metabolism , Muscle, Skeletal/metabolism , Cell Differentiation/genetics , Stem Cells/metabolism , Cells, Cultured , Sciatic NerveABSTRACT
Background: Although the fit-for-purpose approach has been proposed for biomarker assay validation, practical data should be compiled to facilitate the predetermination of acceptance criteria. Methods: Immunoaffinity LC-MS was used to analyze glucagon-like peptide-1 as a model biomarker in six laboratories. Calibration curve, carryover, parallelism, precision, relative accuracy and processed sample stability were evaluated, and their robustness among laboratories was assessed. The rat glucagon-like peptide-1 concentrations in four blinded samples were also compared. Results: The obtained results and determined concentrations in the blinded samples at all laboratories were similar, with a few exceptions, and robust, despite the difference in optimization techniques among laboratories. Conclusion: The results provide insights into the predefinition of the acceptance criteria of immunoaffinity LC-MS-based biomarker assays.
Subject(s)
Laboratories , Tandem Mass Spectrometry , Rats , Animals , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Glucagon-Like Peptide 1 , BiomarkersABSTRACT
Magnetic resonance imaging (MRI) is a more useful diagnostic modality for detecting paranasal tumors compared with computed tomography (CT). We encountered a case of malignant lymphoma of the maxillary sinus. Although CT findings suggested malignancy, MRI findings indicated an inflammatory disease. The patient was a 51-year-old man with a chief complaint of right maxillary toothache. Edema in the right middle meatus and bloody rhinorrhea were observed. CT revealed r ight maxillary s inus shadow with partial bone loss, suggesting malignancy. However, MRI performed two weeks later showed an internal homogeneous lesion with neither contrast effect, nor invasion outside the maxillary sinus. The patient also had no fever, weight loss, or night sweats. Additionally, no palpable cervical lymphadenopathy was observed. Endoscopic sinus surgery was performed to confirm the diagnosis. Upon opening the maxillary sinus, highly viscous retention and a large amount of yellowish-white debris were observed. Allergic fungal rhinosinusitis was suspected. However, histopathological analysis of the debris established a diagnosis of malignant lymphoma. The debris exhibited pathological findings of necrosis. The patient remained in remission after undergoing radiochemotherapy. Malignant lymphomas of the paranasal sinuses, which have a minimal tendency for invasion but with considerable predominance of necrosis, may be diagnosed as an inflammatory disease, based on MRI findings. In cases in which a thorough physical examination could not rule out malignant lymphomas, an endoscopic biopsy should be immediately considered.
Subject(s)
Lymphoma , Male , Humans , Middle Aged , Lymphoma/diagnostic imaging , Lymphoma/pathology , Maxillary Sinus/pathology , Maxillary Sinus/surgery , Tomography, X-Ray Computed , Magnetic Resonance Imaging , NecrosisABSTRACT
Although microsampling of blood is recommended to promote the 3Rs in toxicokinetic (TK) evaluation, there are few reports applying microsampling in actual toxicity evaluation. Here, we assessed the effects of microsampling on toxicological evaluation of methapyrilene hydrochloride, a hepatotoxic substance. Female SD rats received methapyrilene hydrochloride orally at dose levels of 0 (vehicle), 10, and 30 mg/kg BW, once daily for 4 weeks. Each dose level included a microsampling group and a non-microsampling group (n = 5). In the microsampling groups, blood sampling (50 µL/time point) was performed at 6 time points on day 1 of administration and 7 time points on day 27-28; all the animals underwent necropsy on day 29. Toxicity studies and TK analysis were performed, and through these studies in 2 organizations, cross-organization validation of the effect on toxicity evaluation was conducted. In one organization, microsampling obscured changes in some parameters in hematology due to the administration of methapyrilene hydrochloride. In the other organization, although the relationship between the developing pattern of histopathological findings in the liver and the blood sampling was suspected, it was associated with poor reproducibility; this was considered as a change within a variation range of biological reactions. Each of these phenomena was observed in only one organization without consistency. In both organizations, no effect of blood microsampling was observed in other endpoints. In conclusion, microsampling is considered to be a technique applicable to safety studies of drugs showing hepatotoxicity, as it did not show a marked influence on the toxicological evaluation of methapyrilene hydrochloride.
Subject(s)
Drug-Related Side Effects and Adverse Reactions , Methapyrilene , Rats , Female , Animals , Rats, Sprague-Dawley , Methapyrilene/pharmacology , Reproducibility of Results , Blood Specimen Collection/methods , LiverABSTRACT
A palladium-catalysed C-H arylation of benzophospholes with aryl halides has been developed. The reaction with aryl iodides and bromides proceeds well even under phosphine ligand-free Pd(OAc)2 catalysis whereas the Pd(PCy3)2 is effective for the coupling with less reactive aryl chlorides. The optimal conditions are also applicable to the double arylations with organic dihalides and annulation reaction with ortho-dihalogenated benzenes, making the corresponding benzophosphole-based acceptor-donor-acceptor-type molecules and highly condensed heteroacene-type molecules of potent interest in materials chemistry. Although there are many reports of catalytic C-H functionalisations of related benzoheteroles such as indoles, benzothiophenes, and benzofurans, this is the first successful example of the catalytic direct C-H transformation of benzophospholes, to the best of our knowledge. The preliminary optoelectronic properties of some newly synthesized benzophosphole derivatives are also investigated.
ABSTRACT
Drug-induced interstitial lung disease (DILD) occurs when drug exposure causes inflammation of the lung interstitium. DILD can be caused by different types of drugs, and some DILD patterns results in a high mortality rate; hence, DILD poses a serious problem in clinical practice as well as drug development, and strategies to diagnose and distinguish DILD from other lung diseases are necessary. We aimed to identify novel biomarkers for DILD by performing lipidomics analysis on plasma samples from patients with acute and recovery phase DILD. Having identified lysophosphatidylcholines (LPCs) as candidate biomarkers for DILD, we determined their concentrations using validated liquid chromatography/mass spectrometry biomarker assays. In addition, we evaluated the ability of LPCs to discriminate patients with acute phase DILD from those with recovery phase DILD, DILD-tolerant, or other lung diseases, and characterized their association with clinical characteristics. Lipidomics analysis revealed a clear decrease in LPC concentrations in the plasma of patients with acute phase DILD. In particular, LPC(14:0) had the highest discriminative index against recovery phase and DILD-tolerant patients. LPC(14:0) displayed no clear association with causal drugs, or subjects' backgrounds, but was associated with disease severity. Furthermore, LPC(14:0) was able to discriminate between patients with DILD and other lung diseases, including idiopathic interstitial pneumonia and lung disease associated with connective tissue disease. LPC(14:0) is a promising biomarker for DILD that could improve the diagnosis of DILD and help to differentiate DILD from other lung diseases, such as idiopathic interstitial pneumonia and connective tissue disease.
Subject(s)
Connective Tissue Diseases , Idiopathic Interstitial Pneumonias , Lung Diseases, Interstitial , Humans , Lysophosphatidylcholines , Lung Diseases, Interstitial/chemically induced , Lung Diseases, Interstitial/diagnosis , BiomarkersABSTRACT
Background: Many bioanalytical methods for antisense oligonucleotides (ASOs) using LC-MS have been reported. However, no data have been available on the reproducibility and robustness of a single bioanalytical method for ASOs. As such, in the current study, we evaluated the reproducibility and robustness of LC-MS-based bioanalytical methods for ASOs in multiple laboratories. Methods/Results: Seven independent laboratories were included in this study. Mipomersen was measured by ion-pairing LC-MS (IP-LC-MS) as a model ASO using different LC-MS. The validation results of calibration curve, accuracy, precision and selectivity met the criteria of conventional bioanalytical method validation guidelines using LC/GC-MS for drugs in all laboratories. Meanwhile, carryover (>20%) was detected in three laboratories. Conclusion: We first demonstrated the multicenter-validated IP-LC-MS bioanalytical method for ASOs. Our data showed that the method was sensitive, robust and reproducible. However, the occurrence of carryover should be carefully monitored in its future application.
Subject(s)
Biological Therapy , Tandem Mass Spectrometry , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Reproducibility of Results , CalibrationABSTRACT
Among the various histopathological patterns of drug-induced interstitial lung disease (DILD), diffuse alveolar damage (DAD) is associated with poor prognosis. However, there is no reliable biomarker for its accurate diagnosis. Here, we show stratifin/14-3-3σ (SFN) as a biomarker candidate found in a proteomic analysis. The study includes two independent cohorts (including totally 26 patients with DAD) and controls (total 432 samples). SFN is specifically elevated in DILD patients with DAD, and is superior to the known biomarkers, KL-6 and SP-D, in discrimination of DILD patients with DAD from patients with other DILD patterns or other lung diseases. SFN is also increased in serum from patients with idiopathic DAD, and in lung tissues and bronchoalveolar lavage fluid of patients with DAD. In vitro analysis using cultured lung epithelial cells suggests that extracellular release of SFN occurs via p53-dependent apoptosis. We conclude that serum SFN is a promising biomarker for DAD diagnosis.
Subject(s)
Lung Diseases, Interstitial , Pulmonary Surfactant-Associated Protein D , 14-3-3 Proteins , Biomarkers , Exoribonucleases , Humans , Lung Diseases, Interstitial/chemically induced , Lung Diseases, Interstitial/diagnosis , Lung Diseases, Interstitial/pathology , Proteomics , Tumor Suppressor Protein p53ABSTRACT
The coronavirus disease 2019 (COVID-19) pandemic has been associated with high mortality worldwide. Owing to its complicated pathophysiology, diagnostic and prognostic biomarkers for effective patient management remain scarce. We analyzed kynurenine, tryptophan, and serotonin levels in the serum of patients with COVID-19 via liquid chromatography/mass spectrometry analysis. Serum serotonin levels were decreased in patients with more severe COVID-19, along with increased kynurenine and decreased tryptophan concentrations. Patients with moderate disease who subsequently worsened showed significantly lower serotonin concentrations compared with those who did not experience severe disease. Serum serotonin levels may represent a valuable biomarker for COVID-19 severity and prognosis.