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AbstractThe nonspecific binding of peptides to material surfaces is a well-known phenomenon that makes measurement of these substances challenging. Peptide adsorption can be reduced with suitable materials or additives, but the complex nature of biomolecules makes it challenging to find an universal sample preparation method. This study focused on investigating the surface adsorption behavior of motilin to enhance the sensitivity and precision of analytical methods. A simple and specific HPLC method was developed for the quantification of motilin. Using this chromatographic method, it was discovered that motilin binds to both polypropylene and glass surfaces at high levels, with recovery rates of 32 and 24%, respectively. To mitigate this, the effect of protein (bovine serum albumin), salt (sodium chloride), and surfactant (Tween 80) on surface adsorption was investigated. Although all additives reduced adsorption, only Tween 80 achieved nearly 100% recovery with consistent results (RSD ≤2%). Consequently, the sample preparation procedure developed using Tween 80 was integrated into the HPLC method, and the method was validated according to the International Conference on Harmonization guideline [ICH Q2(R1)]. This method is expected to have potential applications in analytical chemistry, drug development, biotechnology, and the pharmaceutical industry for providing accurate and dependable peptide measurements. [ABSTRACT FROM AUTHOR]

Simple Summary: Immune checkpoint inhibitors are used in combination with other drugs as the first-line therapy in the treatment of metastatic renal cell carcinoma. Although these cancer immunotherapies have demonstrated high therapeutic efficacy, half of patients develop treatment resistance. In this study, we investigated the enhancement of the antitumor effect of immunosuppressants by a histone deacetylase (HDAC) inhibitor. A novel HDAC inhibitor, OBP-801, upregulated LMP2, an immunoproteasome subunit, in clear cell renal cell carcinoma (ccRCC) cells, resulting in enhanced MHC class I expression, and a combined treatment of OBP-801 and anti-PD-1 antibody effectively alleviated ccRCC in mice. Our results suggest that manipulating LMP2 expression using OBP-801 may have therapeutic implications for enhancing MHC class I presentation and improving the anti-tumor immune response in ccRCC. In particular, the combination of OBP-801 with anti-PD-1 antibody appears to be promising for ccRCC treatment. Background: Histone deacetylase (HDAC) inhibitors have been reported to exhibit immunomodulatory activities, including the upregulation of major histocompatibility complex class I (MHC class I). Although the immunoproteasome plays a pivotal role in MHC class I antigen presentation, its effect on immunotherapy for clear cell renal cell carcinoma (ccRCC) remains unclear. Methods: This study assessed whether OBP-801, a novel HDAC inhibitor, affects the expression of immunoproteasome subunits and subsequently the MHC class-I-mediated anti-cancer immunity in ccRCC. We analyzed the data of 531 patients with ccRCC from the Cancer Genome Atlas Kidney Clear Cell Carcinoma database. We further evaluated the treatment efficacy of the combination of OBP-801 and anti-PD-1 in a ccRCC mouse model. Results: Low molecular mass polypeptide (LMP) 2 was correlated most positively with CD3E, CD8A, and CD8B expression and estimated CD8+ T cell number. In vitro studies showed that OBP-801 upregulated MHC class I presentation by inducing LMP2 expression in the ccRCC cell lines RENCA, 786-O, and Caki-1. In vivo studies in a syngeneic mouse model with subcutaneous implantation of RENCA cells showed that OBP-801 treatment increased the percentage of CD45+CD3e+ T cells in tumor-infiltrating lymphocytes. The combination of anti-PD-1 antibody and OBP-801 enhanced the anti-tumor effect, LMP2 protein expression, and MHC class I presentation in tumor cells. MHC class I presentation in the tumors of each mouse was positively correlated with the percentage of CD45+CD3e+ T cells. Conclusions: Our results demonstrate that OBP-801 promotes MHC class I presentation through LMP2 upregulation in tumor cells and thereby potentiates PD-1-targeting therapy. These data suggest that the combination of OBP-801 and anti-PD-1 treatment is a promising therapeutic strategy for ccRCC. [ABSTRACT FROM AUTHOR]

Simple Summary: Human epidermal growth factor receptor 2, a protein found on cell surfaces, is often overproduced in aggressive forms of cancers like breast, gastric, ovarian, and lung cancers. This overproduction can lead to rapid tumor growth and poor patient outcomes. Current treatments target this protein to slow cancer progression, but more effective and precise options are needed. This study explores a new approach by designing small protein-like molecules called peptides to specifically block this receptor. Using advanced computer-based methods, researchers identified and optimized several peptides, selecting the ones most likely to bind effectively to this protein. Through further testing, one peptide showed promise as a potential therapy due to its stable binding and favorable properties. This research could lead to new, targeted treatment options for patients with these types of cancers and contribute to ongoing efforts to improve cancer therapies through precision medicine. Background/Objectives: Human epidermal growth factor receptor 2 (HER2) is overexpressed in several malignancies, such as breast, gastric, ovarian, and lung cancers, where it promotes aggressive tumor proliferation and unfavorable prognosis. Targeting HER2 has thus emerged as a crucial therapeutic strategy, particularly for HER2-positive malignancies. The present study focusses on the design and optimization of peptide inhibitors targeting HER2, utilizing machine learning to identify and enhance peptide candidates with elevated binding affinities. The aim is to provide novel therapeutic options for malignancies linked to HER2 overexpression. Methods: This study started with the extraction and structural examination of the HER2 protein, succeeded by designing the peptide sequences derived from essential interaction residues. A machine learning technique (XGBRegressor model) was employed to predict binding affinities, identifying the top 20 peptide possibilities. The candidates underwent further screening via the FreeSASA methodology and binding free energy calculations, resulting in the selection of four primary candidates (pep-17, pep-7, pep-2, and pep-15). Density functional theory (DFT) calculations were utilized to evaluate molecular and reactivity characteristics, while molecular dynamics simulations were performed to investigate inhibitory mechanisms and selectivity effects. Advanced computational methods, such as QM/MM simulations, offered more understanding of peptide–protein interactions. Results: Among the four principal peptides, pep-7 exhibited the most elevated DFT values (−3386.93 kcal/mol) and the maximum dipole moment (10,761.58 Debye), whereas pep-17 had the lowest DFT value (−5788.49 kcal/mol) and the minimal dipole moment (2654.25 Debye). Molecular dynamics simulations indicated that pep-7 had a steady binding free energy of −12.88 kcal/mol and consistently bound inside the HER2 pocket during a 300 ns simulation. The QM/MM simulations showed that the overall total energy of the system, which combines both QM and MM contributions, remained around −79,000 ± 400 kcal/mol, suggesting that the entire protein–peptide complex was in a stable state, with pep-7 maintaining a strong, well-integrated binding. Conclusions: Pep-7 emerged as the most promising therapeutic peptide, displaying strong binding stability, favorable binding free energy, and molecular stability in HER2-overexpressing cancer models. These findings suggest pep-7 as a viable therapeutic candidate for HER2-positive cancers, offering a potential novel treatment strategy against HER2-driven malignancies. [ABSTRACT FROM AUTHOR]

Introduction: Gestational diabetes mellitus (GDM) occurs on the background of increased insulin resistance. We aimed to investigate the levels of plasma pancreastatin (PST) levels and its association with metabolic, demographic, and anthropometric parameters in gestational diabetic and normal glucose-tolerant pregnant women. Materials and methods: A total of 165 pregnant women in the 24th–28th week of pregnancy were enrolled in this cross-sectional study. PST levels were measured using ELISA method. Results: Median PST levels were similar between GDM (n = 62, 37.6%) and normal glucose-tolerant control group (n = 103, 62.4%). In the GDM group, PST level showed a significant positive correlation with fasting insulin and the homeostasis model assessment of insulin resistance (HOMA-IR). In binary logistic regression analysis, PST levels did not predict the presence of GDM significantly. Conclusion: Positive correlation of PST with fasting insulin and HOMA-IR suggests that PST could be associated with insulin resistance. Further studies are needed as regards to the role of PST in GDM pathogenesis. [ABSTRACT FROM AUTHOR]

Backgrounds: Various physiological functions and reaction cascades, as well as disease progression in the living systems, are controlled by the activity of specific proteolytic enzymes. We conducted the study to evaluate protease activity by assessing peptide fragments from either conserved or labeled red blood cells (RBCs) with aminofluorescein (AF) in the reaction media. Methods: RBCs were incubated in media containing trypsin. Subsequently, the concentration of peptide fragments in the reaction media, resulted by the digestion with trypsin from conserved cells, was estimated by 3‐(4‐carboxybenzoyl)quinoline‐2‐carboxaldehyde (CBQCA) as an amine‐reactive fluorogenic reagent. In a second approach, we conjugated AF to the conserved RBCs and then exposed AF‐labeled RBCs to trypsin. This was followed by directly measuring the fluorescence intensity (FI) in the reaction media to estimate the concentration of AF‐labeled peptide fragments resulting from the enzyme's activity. Results: Show a concentration‐ and time‐dependent increase in FIs, reflecting the activity of trypsin as a proteolytic enzyme. The FIs increased significantly by 4 to 5 folds in samples treated with different enzyme concentrations, and by over 11 folds after 2 h incubation in media containing a 50 μL trypsin, as evidenced by CBQCA assays. Conclusion: These fast and affordable approaches could be applied with high reliability for the general estimation of protease activity in samples and customized for diagnostic purposes and prognostic evaluation in various diseases. [ABSTRACT FROM AUTHOR]

Objective: Interstitial lung diseases (ILDs) are a heterogeneous group of disorders that can develop in patients with connective tissue diseases. Establishing autoimmunity in ILD impacts prognosis and treatment. Patients with ILD are screened for autoimmunity by measuring antinuclear autoantibodies, rheumatoid factors, and other nonspecific tests. However, this approach may miss autoimmunity that manifests as autoantibodies to tissue antigens not previously defined in ILD. Methods: We use Phage Immunoprecipitation‐Sequencing (PhIP‐Seq) to conduct an autoantibody discovery screen of patients with ILD and controls. We screened for novel autoantigen candidates using PhIP‐Seq. We next developed a radio‐labeled binding assay and validated the leading candidate in 398 patients with ILD recruited from two academic medical centers and 138 blood bank individuals that formed our reference cohort. Results: PhIP‐Seq identified 17 novel autoreactive targets, and machine learning classifiers derived from these targets discriminated ILD serum from controls. Among the 17 candidates, we validated CDHR5 and found CDHR5 autoantibodies in patients with rheumatologic disorders and importantly, patients not previously diagnosed with autoimmunity. Using survival and transplant free–survival data available from one of the two centers, patients with CDHR5 autoantibodies showed worse survival compared with other patients with connective tissue disease ILD. Conclusion: We used PhIP‐Seq to define a novel CDHR5 autoantibody in a subset of select patients with ILD. Our data complement a recent study showing polymorphisms in the CDHR5‐IRF7 gene locus strongly associated with titer of anticentromere antibodies in systemic sclerosis, creating a growing body of evidence suggesting a link between CDHR5 and autoimmunity. [ABSTRACT FROM AUTHOR]

Along with the recent acknowledgement of the World Anti‐Doping Agency to use dried blood spot (DBS) samples for routine doping control purposes, there have been propositions to use DBS as a matrix that allows regular proactive remotely supervised self‐sampling, providing potential longitudinal monitoring of an athlete's exposure to doping agents. However, several organizational aspects have to be considered before implementation, such as the verification of the sample collections time point. Based on a previous untargeted proteomics workflow utilizing liquid chromatography–high‐resolution mass spectrometry (LC–HRMS) to identify protein/peptide markers to define the time since deposition of a bloodstain, the aim of the current study was to develop a targeted LC–HRMS/MS analytical method for promising peptidic target analytes. A long‐term DBS storage experiment was carried out over a 3‐month period (sample collection time points: 0, 2, 4, 7, 14, 21, 28, 42, 56, 70, 84 and 91 days) with DBS samples of 10 volunteers for longitudinal investigation of signal abundance changes of targeted peptide sequences at different storage temperatures (room temperature [RT], 4°C and −20°C). Prior to experimental analysis, LC–HRMS/MS method characteristics were successfully assessed, including intraday precision, carryover and sample extract stability. For estimation of DBS sample collection time points, ratios of two peptides that originate from the same protein prior to tryptic digestion were created. Two targeted peptide area ratios were found to significantly increase after being stored at RT for 28 days, representing potential markers for future use in routine doping controls that contribute to advancing complementary avenues in anti‐doping. [ABSTRACT FROM AUTHOR]

The development of selective and sensitive probes for monitoring caspase-3 activity-a critical enzyme involved in apoptosis-remains an area of significant interest in biomedical research. Herein, we report the synthesis and characterisation of a novel ratiometric fluorescent probe, Ac-DEVD-PABCNaph, designed to detect caspase-3 activity. The probe utilises a 1,8-naphthalimide fluorophore covalently linked to a peptide sequence via a self-immolative p-aminobenzyl alcohol (PABA) linker. Upon enzymatic cleavage by caspase-3, the probe undergoes spontaneous degradation, releasing the free naphthalimide fluorophore, resulting in a ratiometric change in fluorescence emission. Spectroscopic studies revealed a time-dependent ratiometric fluorescent response, demonstrating the probe's ability to visualise caspase-3 activity with high sensitivity. Enzyme kinetics such as Km (Michaelis constant), kcat (turnover number), and LOD (Limit of Detection) were obtained, suggesting that the probe possesses comparable kinetic data to other probes in literature, but with the added benefits of ratiometric detection. Selectivity studies also demonstrated the probe's specificity for caspase-3 over other endogenous species and enzymes. Ac-DEVD-PABC-Naph may be a promising tool for the quantitative detection and fluorescent visualisation of caspase-3 activity in biological systems, with potential applications in apoptosis research and drug development. [ABSTRACT FROM AUTHOR]

Objective: Opiorphin is a pentapeptide secreted in saliva and has a strong analgesic effect. Salivary opiorphin has been shown to increase in orofacial pain and may act as a pain reliever in pain caused by denture‐related ulcers. The current study aimed to evaluate the salivary opiorphin levels in traumatic ulcers caused by ill‐fitting dentures and demonstrate whether there is any correlation between trauma‐related pain levels and salivary opiorphin levels. Materials and Methods: Twenty‐two individuals with new full dentures and a complaint of pain due to ill‐fitting were included in this study. Patients were asked to rate their level of pain on a visual analog scale (VAS). Then, saliva specimens were collected at the first visit with the complaint of pain and 7 days following the denture adjustment. Results: The average saliva opiorphin level before and after denture adjustment were 19.29 ± 5.44 and 15.78 ± 3.95 ng/mL, respectively. A dependent (paired) t‐test determined that the mean salivary opiorphin level differed statistically significantly before and after the adjustment of the dentures. Conclusions: The findings show that salivary opiorphin levels increase in pain associated with denture‐related traumatic ulcers. Adjusting the dentures resulted in pain relief and a statistically significant reduction in opiorphin levels. [ABSTRACT FROM AUTHOR]

Introduction: Human leukocyte antigen (HLA)-DPB1 mismatches during hematopoietic stem cell transplantation (HSCT) with an unrelated donor result in an increased risk for the development of graft-versus-host disease (GvHD). The number of CD8+ T-cell epitopes available for indirect allorecognition as predicted by the PIRCHE algorithm has been shown to be associated with GvHD development. As a proof of principle, PIRCHE-I predictions for HLA-DPB1 mismatches were validated in vitro and in vivo. Methods: PIRCHE-I analysis was performed to identify HLA-DPB1-derived peptides that could theoretically bind to HLA-A*02:01. PIRCHE-I predictions for HLA-DPB1 mismatches were validated in vitro by investigating binding affinities of HLA-DPB1-derived peptides to the HLA-A*02:01 in a competition-based binding assay. To investigate the capacity of HLA-DPB1-derived peptides to elicit a T-cell response in vivo, mice were immunized with these peptides. T-cell alloreactivity was subsequently evaluated using an interferon-gamma ELISpot assay. Results: The PIRCHE-I algorithm identified five HLA-DPB1-derived peptides (RMCRHNYEL, YIYNREEFV, YIYNREELV, YIYNREEYA, and YIYNRQEYA) to be presented by HLA-A*02:01. Binding of these peptides to HLA-A*02:01 was confirmed in a competition-based peptide binding assay, all showing an IC50 value of 21 μm or lower. The peptides elicited an interferon-gamma response in vivo. Conclusion: Our results indicate that the PIRCHE-I algorithm can identify potential immunogenic HLA-DPB1-derived peptides present in recipients of an HLA-DPB1-mismatched donor. These combined in vitro and in vivo observations strengthen the validity of the PIRCHE-I algorithm to identify HLA-DPB1 mismatch-related GvHD development upon HSCT. [ABSTRACT FROM AUTHOR]

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The new natural products reported in 2022 in peer-reviewed articles in journals with good reputations were reviewed and analyzed. The advances made by Asian research groups in the field of natural products chemistry in 2022 were summarized. Compounds with unique structural features and/or promising bioactivities originating from Asian natural sources were discussed based on their structural classification. [ABSTRACT FROM AUTHOR]

This study investigated sex differences in Fe status, and associations between Fe status and endurance and musculoskeletal outcomes, in military training. In total, 2277 British Army trainees (581 women) participated. Fe markers and endurance performance (2·4 km run) were measured at the start (week 1) and end (week 13) of training. Whole-body areal body mineral density (aBMD) and markers of bone metabolism were measured at week 1. Injuries during training were recorded. Training decreased Hb in men and women (mean change (–0·1 (95 % CI –0·2, –0·0) and –0·7 (95 % CI –0·9, –0·6) g/dl, both P < 0·001) but more so in women (P < 0·001). Ferritin decreased in men and women (–27 (95 % CI –28, –23) and –5 (95 % CI –8, –1) µg/l, both P ≤ 0·001) but more so in men (P < 0·001). Soluble transferrin receptor increased in men and women (2·9 (95 % CI 2·3, 3·6) and 3·8 (95 % CI 2·7, 4·9) nmol/l, both P < 0·001), with no difference between sexes (P = 0·872). Erythrocyte distribution width increased in men (0·3 (95 % CI 0·2, 0·4)%, P < 0·001) but not in women (0·1 (95 % CI –0·1, 0·2)%, P = 0·956). Mean corpuscular volume decreased in men (–1·5 (95 % CI –1·8, –1·1) fL, P < 0·001) but not in women (0·4 (95 % CI –0·4, 1·3) fL, P = 0·087). Lower ferritin was associated with slower 2·4 km run time (P = 0·018), sustaining a lower limb overuse injury (P = 0·048), lower aBMD (P = 0·021) and higher beta C-telopeptide cross-links of type 1 collagen and procollagen type 1 N-terminal propeptide (both P < 0·001) controlling for sex. Improving Fe stores before training may protect Hb in women and improve endurance and protect against injury. [ABSTRACT FROM AUTHOR]

Background: Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) is involved in the stress response and may play a key role in mood disorders, but no information is available on PACAP for the human brain in relation to mood disorders. Methods: PACAP-peptide levels were determined in a major stress-response site, the hypothalamic paraventricular nucleus (PVN), of people with major depressive disorder (MDD), bipolar disorder (BD) and of a unique cohort of Alzheimer's disease (AD) patients with and without depression, all with matched controls. The expression of PACAP-(Adcyap1mRNA) and PACAP-receptors was determined in the MDD and BD patients by qPCR in presumed target sites of PACAP in stress-related disorders, the dorsolateral prefrontal cortex (DLPFC) and anterior cingulate cortex (ACC). Results: PACAP cell bodies and/or fibres were localised throughout the hypothalamus with differences between immunocytochemistry and in situ hybridisation. In the controls, PACAP-immunoreactivity-(ir) in the PVN was higher in women than in men. PVN-PACAP-ir was higher in male BD compared to the matched male controls. In all AD patients, the PVN-PACAP-ir was lower compared to the controls, but higher in AD depressed patients compared to those without depression. There was a significant positive correlation between the Cornell depression score and PVN-PACAP-ir in all AD patients combined. In the ACC and DLPFC, alterations in mRNA expression of PACAP and its receptors were associated with mood disorders in a differential way depending on the type of mood disorder, suicide, and psychotic features. Conclusion: The results support the possibility that PACAP plays a role in mood disorder pathophysiology. [ABSTRACT FROM AUTHOR]

Simple Summary: Colorectal cancer is a leading cause of worldwide cancer-related morbidity and mortality. Conventional white light colonoscopy is routinely used to identify and remove colonic polyps. However, premalignant lesions that are flat and subtle in appearance are often missed. Cells alter in molecular expression well in advance of morphological changes. cMet is highly overexpressed by colonic adenomas. We aim to implant patient-derived organoids in the colon of immunocompromised mice to demonstrate a model system for evaluating the specific uptake of a near-infrared labeled peptide ligand. Human adenoma and normal colonoids were imaged in vivo using a small animal endoscope. A high target-to-background ratio was observed ~1 h post-injection followed by clearance after ~24 h. The mean intensity measured was significantly greater for the adenoma versus normal colonoids. These results demonstrate promise for a targeted imaging approach using fluorescently labeled peptides to identify precancerous lesions that would otherwise be missed using white light illumination alone. Colorectal cancer is a leading cause of cancer-related morbidity and mortality worldwide. Premalignant lesions that are flat and subtle in morphology are often missed in conventional colonoscopies. Patient-derived adenoma colonoids with high and low cMet expression and normal colonoids were implanted orthotopically in the colon of immunocompromised mice to serve as a preclinical model system. A peptide specific for cMet was labeled with IRDye800, a near-infrared (NIR) fluorophore. This peptide was administered intravenously, and in vivo imaging was performed using a small animal fluorescence endoscope. Quantified intensities showed a peak target-to-background ratio at ~1 h after intravenous peptide injection, and the signal cleared by ~24 h. The peptide was stable in serum with a half-life of 3.6 h. Co-staining of adenoma and normal colonoids showed a high correlation between peptide and anti-cMet antibody. A human-specific cytokeratin stain verified the presence of human tissues implanted among surrounding normal mouse colonic mucosa. Peptide biodistribution was consistent with rapid renal clearance. No signs of acute toxicity were found on either animal necropsy or serum hematology and chemistries. Human colonoids provide a clinically relevant preclinical model to evaluate the specific uptake of a NIR peptide to detect premalignant colonic lesions in vivo. [ABSTRACT FROM AUTHOR]

Two new linear peptides, penicamides A and B (1 and 2), together with four known analogous were isolated from the extracts of the marine-derived fungus Penicillium sp. SCSIO 41512. Their structures were elucidated by analysis of 1D/2D NMR data and HRESI-MS. Their absolute configurations were established by Marfey's methods and quantum chemical calculations. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to investigate the association of GCF TREM‐1, PGLYRP1, and IL‐1β levels with periodontal health in pre‐ and postmenopausal women. Background: Triggering receptor expressed on myeloid cells 1 (TREM‐1), activated through its ligand peptidoglycan recognition protein 1 (PGLYRP1), stimulates proinflammatory cytokine production, such as interleukin (IL)‐1β, during periodontal inflammation. Postmenopausal changes may modulate these immune‐inflammatory functions. No clinical study has yet investigated the effect of menopause on TREM‐1, PGLYRP1, and IL‐1β levels in gingival crevicular fluid (GCF). Methods: This cross‐sectional study included 148 women (age range = 35–65 years), divided into postmenopausal women (PMW) (n = 76, mean age = 54 ± 5 years) and regularly menstruating premenopausal women (RMPW) (n = 72, mean age = 40 ± 4 years). Clinical periodontal parameters were recorded. TREM‐1, PGLYRP1, and IL‐1β levels were quantified with enzyme‐linked immunosorbent assays. Pearson's Chi‐squared test and Mann–Whitney‐U test were used to compare categorical and numerical variables, respectively. Spearman's Rho correlation analysis was used to test the linear relationship between variables. Analyte level data were categorized based on the periodontal diagnosis and menopause status (2 × 2 nonparametric factorial ANOVA). Results: No significant differences in TREM‐1, PGLYRP1, and IL‐1β levels between PMW and RMPW were observed (p >.05). Mean values of periodontal indexes including probing depth did not differ significantly between PMW and RMPW groups (p =.474). TREM‐1 levels were significantly higher in both PMW and RMPW with periodontitis, compared to gingivitis or health (p =.0021). Conclusion: Menopause‐related changes have no observable effect on GCF levels of TREM‐1, PGLYRP1, and IL‐1β. Higher GCF TREM‐1 levels in women with periodontitis regardless of their menopausal status indicate that TREM‐1 may be an indicator for periodontitis both in premenopausal and postmenopausal women. [ABSTRACT FROM AUTHOR]

Objective: To examine the anti-inflammatory effects and potential mechanisms of polypeptide from Moschus (PPM) in lipopolysaccharide (LPS)-induced THP-1 macrophages and BALB/c mice. Methods: The polypeptide was extracted from Moschus and analyzed by high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Subsequently, LPS was used to induce inflammation in THP-1 macrophages and BALB/c mice. In LPS-treated or untreated THP-1 macrophages, cell viability was observed by cell counting kit 8 and lactate dehydrogenase release assays; the proinflammatory cytokines and reactive oxygen species (ROS) were measured by enzyme-linked immunosorbent assay and flow cytometry, respectively; and protein and mRNA levels were measured by Western blot and real-time quantitative polymerase chain reaction (qRT-PCR), respectively. In LPS-induced BALB/c mice, the proinflammatory cytokines were measured, and lung histology and cytokines were observed by hematoxylin and eosin (HE) and immunohistochemical (IHC) staining, respectively. Results: The SDS-PAGE results suggested that the molecular weight of purified PPM was in the range of 10–26 kD. In vitro, PPM reduced the production of interleukin 1β (IL-1β), IL-18, tumor necrosis factor α (TNF-α), IL-6 and ROS in LPS-induced THP-1 macrophages (P<0.01). Western blot analysis demonstrated that PPM inhibited LPS-induced nuclear factor κB (NF-κB) pathway and thioredoxin interacting protein (TXNIP)/nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain containing 3 (NLRP3) inflammasome pathway by reducing protein expression of phospho-NF-κB p65, phospho-inhibitors of NF-κB (Iκ Bs) kinase α/β (IKKα/β), TXNIP, NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), and pro-caspase-1 (P<0.05 or P<0.01). In addition, qRT-PCR revealed the inhibitory effects of PPM on the mRNA levels of TXNIP, NLRP3, ASC, and caspase-1 (P<0.05 or P<0.01). Furthermore, in LPS-induced BALB/c mice, PPM reduced TNF-α and IL-6 levels in serum (P<0.05 or P<0.01), decreased IL-1β and IL-18 levels in the lungs (P<0.01) and alleviated pathological injury to the lungs. Conclusion: PPM could attenuate LPS-induced inflammation by inhibiting the NF-κB-ROS/NLRP3 pathway, and may be a novel potential candidate drug for treating inflammation and inflammation-related diseases. [ABSTRACT FROM AUTHOR]

Background. A major issue is finding a valid biomarker able to diagnose periodontal disease with the presence and absence of risk factors. Indeed, the association between smoking and periodontal diseases and its impact on the manifestation of antimicrobial peptides has been delineated in clinical and epidemiological investigations. The antimicrobial peptide cathelicidin (LL-37) is pivotal in preserving periodontal health. Objectives. This investigation examines and contrasts the levels of cathelicidin in the saliva of smokers and nonsmokers of periodontitis. The study also seeks to establish this biomarker's diagnostic ability to differentiate between periodontal health and disease. Materials and Methods. The study involved the collection of unstimulated saliva samples from 160 participants, comprising 80 patients diagnosed with periodontitis (40 of whom were smokers and 40 were nonsmokers) and 80 periodontitis-free individuals (40 smokers and 40 nonsmokers). The clinical periodontal parameters were assessed, including recording the probing pocket depth, the level of clinical attachment, and the percentage of bleeding on probing. Subsequently, enzyme-linked immunosorbent assays were conducted to quantify the protein levels of LL-37 in the saliva samples obtained from the subjects mentioned above. Results. The highest level of salivary LL-37 was found in the nonsmoker periodontitis (NSP) patients, followed by the group of smoker periodontitis (SP) and then nonsmoker healthy (NSH) group, while the lowest level was found in the healthy smoker (HS) group. At the same time, the LL-37 seems to be a very good biomarker in differentiating periodontal health from disease with the presence and absence of smoking. Conclusion. Periodontitis results in a significant elevation of salivary LL-37 levels in smoker and nonsmoker patients compared to healthy individuals. These levels are positively correlated with the periodontal parameter and can serve as a valuable diagnostic tool to predict periodontitis, whereas smoking significantly reduces these levels. [ABSTRACT FROM AUTHOR]

Immunopeptidomics has made tremendous contributions to our understanding of antigen processing and presentation, by identifying and quantifying antigenic peptides presented on the cell surface by Major Histocompatibility Complex (MHC) molecules. Large and complex immunopeptidomics datasets can now be routinely generated using Liquid Chromatography-Mass Spectrometry techniques. The analysis of this data – often consisting of multiple replicates/conditions – rarely follows a standard data processing pipeline, hindering the reproducibility and depth of analysis of immunopeptidomic data. Here, we present Immunolyser, an automated pipeline designed to facilitate computational analysis of immunopeptidomic data with a minimal initial setup. Immunolyser brings together routine analyses, including peptide length distribution, peptide motif analysis, sequence clustering, peptide-MHC binding affinity prediction, and source protein analysis. Immunolyser provides a user-friendly and interactive interface via its webserver and is freely available for academic purposes at https://immunolyser.erc.monash.edu/. The open-access source code can be downloaded at our GitHub repository: https://github.com/prmunday/Immunolyser. We anticipate that Immunolyser will serve as a prominent computational pipeline to facilitate effortless and reproducible analysis of immunopeptidomic data.

Background: The free‐access (FA) intravenous alcohol self‐administration (IV‐ASA) paradigm is an experimental approach that can identify modulators of alcohol consumption in humans. Moreover, the outcome measures of IV‐ASA paradigms are associated with self‐reported alcohol intake using the timeline follow‐back method (TLFB). To evaluate how FA IV‐ASA reflects drinking in real life, we examined the relationship between an objective marker of recent alcohol intake, phosphatidylethanol in blood (B‐PEth), and TLFB and measures obtained during IV‐ASA in individuals with alcohol use disorder (AUD) and social drinkers (SD). We also explored the associations between these measures and gut‐brain peptides involved in AUD pathophysiology. Methods: Thirty‐eight participants completed a laboratory session in which they self‐administered alcohol intravenously. The safety limit was 200 mg%, and main outcomes were mean and peak breath alcohol concentrations (BrAC). Blood samples were drawn prior to IV‐ASA and subjective alcohol effects were rated during the experiment. Results: The study sample comprised 24 SD and 14 participants with DSM‐5 mild AUD. Although BrACs were not associated with B‐PEth or TLFB in the full sample or AUD subgroup, there was an association with TLFB in SD. In both subgroups, BrACs were associated with alcohol craving but with differential timing. Total ghrelin levels were higher in AUD participants than in SD. Conclusions: No associations between B‐PEth levels and achieved BrACs were observed in the mild AUD group, the SD group, or the full sample. The ability for FA IV‐ASA to reflect recent drinking was confirmed only for TLFB in SD, whereas there were no associations within the smaller subsample of participants with mild AUD or in the full sample. Further studies that include a larger AUD sample are warranted. The association of BrACs with craving for alcohol suggests that the IV‐ASA method may be useful for assessing interventions that target craving. This could be explored by using the FA IV‐ASA model to evaluate the effects on craving of approved pharmacotherapies for AUD. [ABSTRACT FROM AUTHOR]

BACKGROUND: Cyanobacterial harmful algal blooms (CyanoHABs) originate from the excessive growth or bloom of cyanobacteria often referred to as blue-green algae. They have been on the rise globally in both marine and freshwaters in recently years with increasing frequency and severity owing to the rising temperature associated with climate change and increasing anthropogenic eutrophication from agricultural runoff and urbanization. Humans are at a great risk of exposure to toxins released from CyanoHABs through drinking water, food, and recreational activities, making CyanoHAB toxins a new class of contaminants of emerging concern. OBJECTIVES: We investigated the toxic effects and mechanisms of microcystin-LR (MC-LR), the most prevalent CyanoHAB toxin, on the ovary and associated reproductive functions. METHODS: Mouse models with either chronic daily oral or acute intraperitoneal exposure, an engineered three-dimensional ovarian follicle culture system, and human primary ovarian granulosa cells were tested with MC-LR of various dose levels. Single-follicle RNA sequencing, reverse transcription– quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, western blotting, immunohistochemistry (IHC), and benchmark dose modeling were used to examine the effects of MC-LR on follicle maturation, hormone secretion, ovulation, and luteinization. RESULTS: Mice exposed long term to low-dose MC-LR did not exhibit any differences in the kinetics of folliculogenesis, but they had significantly fewer corpora lutea compared with control mice. Superovulation models further showed that mice exposed to MC-LR during the follicle maturation window had significantly fewer ovulated oocytes. IHC results revealed ovarian distribution of MC-LR, and mice exposed to MC-LR had significantly lower expression of key follicle maturation mediators. Mechanistically, in both murine and human granulosa cells exposed to MC-LR, there was reduced protein phosphatase 1 (PP1) activity, disrupted PP1-mediated PI3K/AKT/FOXO1 signaling, and less expression of follicle maturation-related genes. DISCUSSION: Using both in vivo and in vitro murine and human model systems, we provide data suggesting that environmentally relevant exposure to the CyanoHAB toxin MC-LR interfered with gonadotropin-dependent follicle maturation and ovulation. We conclude that MC-LR may pose a nonnegligible risk to women’s reproductive health by heightening the probability of irregular menstrual cycles and infertility related to ovulatory disorders. [ABSTRACT FROM AUTHOR]

Sensory hair cells are mechanoreceptors required for hearing and balance functions. Stereociliary bundles play a critical role in mechanoelectrical transduction (MET), many prior studies examining developing and regenerated hair cells have assessed bundle morphology to determine cell maturity. However, only few studies have systematically assessed hair bundle dimensions during embryonic and postnatal development. In this study, utricles were collected from embryonic (E) 13.5, 15.5, E18.5, postnatal day (P) 0, P37 and P180 wild type mice, and immunostained with the kinocilia marker α-Tubulin and stereocilia marker phalloidin. From 3D reconstructed images of hair cells, kinocilia height, tallest and shortest cilia, and volume were measured. At E13.5, height of kinocilia, tallest and shortest cilia were relatively uniform. Starting at E15.5, wide distributions of hair bundle and kinocilia heights were observed, possibly because of maturing kinocilia and bundles in older hair cells and the emergence of hair bundles and kinocilia in new hair cells in late embryonic and early postnatal periods. Some longer kinocilia appeared curled or bent. Short bundles were still observed in adult mouse utricle. Expression of the actin-crosslinking protein FSCN2 was also examined to characterize the maturity of hair bundles. FSCN2 was absent at E13.5 and E15.5, became detectable in most hair bundles by E18.5 and P0, but some short bundles still lacked FSCN2 even at 6 months. To mark newly born hair cells, we fate-mapped supporting cells in Plp1CreERT/+; Rosa26RtdTomato/+ mice by treating them with tamoxifen at P3. At 1, 2 and 6 months, most bundles of traced hair cells still appeared relatively shorter than untraced ones, and continue to display short, curled or bent kinocilia, resembling those in the early embryonic stages. By 6 months, bundles of only 46% of traced hair cells expressed FSCN2, compared to 97% of the untraced hair cells. Together, our data indicate that hair cells in the embryonic utricle display short kinocilia and stereocilia that elongate over time. Newly added hair cells display short bundle and kinocilia that resemble those during embryonic periods, with some remain detectable in the adult utricle. [ABSTRACT FROM AUTHOR]

Background: Apitherapy is an emerging field in cancer research, particularly in developing communities. The potency of Melittin (MEL), a major constituent in bee venom is accounted for the cytotoxic capacity against cancer cells. It is postulated that the genotype of bees and the time of venom collection influences its specific activity against certain types of cancer. Method: Hereby, Jordanian crude bee venom (JCBV) was collected during different seasons of the year, specifically spring, summer and autumn and investigated for in vitro antitumour effects. Venom collected during springtime comprised the highest quantity of MEL in comparison to venom collected some other time. Springtime-collected JCBV extract and MEL were tested on an immortal myelogenous leukaemia cell line, namely K562 leukemic cells. Treated cells were examined for cell modality via flow cytometry analysis and cell death mediating gene expressions. Results: Springtime-collected JCBV extract and MEL showed an IC50 of 3.7 ± 0.37 μg/ml and 1.84 ± 0.75 μg/ml, respectively. In comparison to JCBV and positive control, MEL-treated cells exhibited late apoptotic death with a moderate cellular arrest at G0/G1 and an increase of cell number at G2/M phase. Expression of NF-κB/MAPK14 axis was inhibited in MEL and JCBV-treated cells, as well as expression of c-MYC and CDK4. Moreover, marked upregulation in ABL1, JUN and TNF was observed. In conclusion, springtime-collected JCBV showed the highest content of MEL while both JCBV and pure MEL showed apoptotic, necrotic, and cell cycle arrest efficiency against K562 leukemic cells. Conclusion: Integration of bee venom in chemotherapy needs more investigation and should be carefully translated into clinical use. During such translation, the correlation of bee genotype, collection time and concentration of MEL in CBV should be profiled. [ABSTRACT FROM AUTHOR]

Although Blufensins (Bln) have important functions in the response of plants to biotic stress the precise functioning of Bln in wheat remains largely unknown. Here we isolated a Bln gene (TaBln4) from Suwon 11 infected by Puccinia striiformis f. sp. tritici (Pst). Expression of TaBln4 increased in host plants at the early stage of infection with a virulent Pst race (CYR31) but was unchanged in response to infection by an avirulent race (CYR23). Transcription levels of TaBln4 were also regulated by hormone and abiotic stresses. Expression of TaBln4 in tobacco leaves suppressed Bax-induced programmed cell death. Knockdown of TaBln4 by virus-induced gene silencing inhibited colonization of race CYR31 by increasing the accumulation of H2O2 and formation of hypersensitive responses (HR). Transient overexpression of TaBln4 by a transient overexpression system (BSMV-VOX) increased the susceptibility of wheat to CYR31. Results from bimolecular fluorescence complementation and pull-down assays demonstrated that TaBLN4 interacted with calmodulin. Taken together, our results suggest that TaBln4 negatively regulates resistance in wheat to Pst in a reactive oxygen species (ROS)- and HR-dependent manner. [ABSTRACT FROM AUTHOR]

Vitamin D is an important nutrient in the body that plays a vital role in immune system function. Several epidemiologic studies have shown that low vitamin D levels are found in a large percentage of COVID-19 patients with acute respiratory failure and that vitamin D levels may predict mortality in COVID-19 infection. Based on these findings, vitamin D supplementation may be an effective approach to preventing and/or treating COVID-19. Potential underlying mechanisms and clinical trial data evaluating the impact of supplementation in humans are described below. [ABSTRACT FROM AUTHOR]

Medicinal plants and their consituents play a crucial role in disease prevention and treatment. The genus Gypsophila (Caryophyllaceae family), comprising approximately 150 species of flowering plants, holds significant value in both ornamental and therapeutic fields. Traditional uses in various cultures highlight their potential in treating a range of conditions, such as liver disorders, diabetes, and kidney stones. These resilient plants, known for their delicate blooms and adaptability to diverse environments, are rich in chemical compounds, including non-volatile constituents such as phenolic compounds (e.g., flavonoids), terpenoids, saponins, cyclopeptides, and alkaloids. Additionally, monoterpenes and sesquiterpenes, the primary volatile constituents, exhibit significant insecticidal properties. Gypsophila species show a broad spectrum of pharmacological effects, including hepatoprotective, cytotoxic, anti-diabetic, antioxidant, cytotoxic, anti-inflammatory, diuretic, neuroprotective, and anti-obesity properties. This review underscores the promising therapeutic potential of Gypsophila and advocates for further research, particularly clinical trials, to thoroughly assess their efficacy and safety. By consolidating existing knowledge, it sheds light on the potential of Gypsophila species as valuable resources for human health and underscores the need for continued exploration to uncover novel treatments and interventions. [Display omitted] [ABSTRACT FROM AUTHOR]

Integerrimide-A (IG-A) is a cyclic heptapeptide that was recently synthesized after being recovered from the latex of the Jatropha integerrima tree. This was achieved by first coupling a tetrapeptide unit (Boc-Gly-L-Leu-L-Leu-L-Leu-OMe) with a tripeptide unit (L-Thr-L-Pro-L-Trp-OMe). The characterization was done by using spectral techniques like FT-IR, 1H-NMR, mass spectrometry, and elemental analysis of the newly synthesized cyclic molecule. Antimicrobial and anticancer properties of IG-A were tested using a biological screening. Gram +ve bacteria (B. subtilis and S. aureus) and Gram -ve bacteria (P. aeruginosa and E. coli) were used in the antibacterial testing. Fungal strains such as C. albicans, A. niger, T. mentagrophytes, and M. audouinii were used to test the antifungal activities. Antimicrobial activity analysis revealed that cyclic peptide—IG-A (8)—has modest antibacterial activity and antifungal activities, when compared with the standard drugs ciprofloxacin and griseofulvin, respectively. Comparable MTT assays were performed on HCT116 (human colon carcinoma) and B16F10 (melanoma cells) cell lines with doxorubicin as the standard drug to determine the cytotoxic activity of the synthesized cyclic peptide. Inhibition of growth of HCT116 and B16F10 cell lines was used to calculate the cytotoxic effect. At a dosage of 120 μg/mL, the cyclopeptide IG-A (8) inhibited cell proliferation by 87.5 and 72.5 percent, respectively. Cyclopeptide IG-A had CTC50 values of 77.65 μM and 68.63 μM against HCT116 and B16F10, respectively. The % growth inhibitions at lesser levels are 72.5 and 50 at 60 μg/mL, respectively. The standard drug inhibited growth by 100 percent with CTC50 values of 48.63 μM and 43.25 μM against HCT116 and B16F10, respectively. From the results, it is concluded that IG-A has considerable antimicrobial and cytotoxic effects. Internucleosomal DNA fragmentation may be the underlying mechanism in HCT116 cells, whereas the suppression of eumelanin synthesis in B16F10 cells is another possibility. [ABSTRACT FROM AUTHOR]

Enzymatic hydrolysis of hen egg confers beneficial functionalities, but it is essential to control the progress of hydrolysis to enhance the quality and reduce allergy risk. However, a reliable method for directly controlling the progress of egg protein hydrolysis is unavailable. Focusing on ovalbumin, a major protein and allergen in egg white, we demonstrated the applicability of liquid chromatography–tandem mass spectrometry (LC–MS/MS) to assess the degradation of ovalbumin in its enzymatic hydrolysates. Using immobilized Alcalase or Everlase, the degradation degree (DD) of ovalbumin was determined using LC–MS/MS; the result (8.8–95.3%) was highly correlated with the DD (8.9–92.3%) obtained using size-exclusion chromatography (SEC-HPLC) (r > 0.96, p < 0.001). In egg white hydrolysates with a DD of 16.1–99.5%, ovalbumin recovery when spiked with 1250 ng/mL was 81.1–114.8% with excellent dilution linearity (r2 > 0.99). These results indicate that LC–MS/MS is a reliable method for increasing the quality of enzymatically hydrolyzed egg products for food, nutraceutical, pharmaceutical, and cosmetic industries. [ABSTRACT FROM AUTHOR]

Marine biofouling causes serious harms to surfaces of marine devices in transportation, aquaculture, and offshore construction. Traditional antifouling methods pollute the environment. A novel and green antifouling strategy was developed to prevent effectively the adhesion of bacteria and microalgae. An antifouling surface was fabricated via coating Turgencin BMox2 (TB) onto dopamine-modified 304 stainless steel (304 SS). The surface physical and chemical properties before and after modification were characterized by Fourier transform infrared spectrometer (FTIR), X-ray photoelectron spectrometer (XPS), contact angle measurement (CA), 3D optical profilometer, ellipsometer, and atomic force microscope (AFM). Antimicrobial peptide was coated onto the surface of 304 SS successfully, and the surface morphology and wettability of the modified sample were modified. Moreover, cytocompatibility of the peptide was evaluated by co-culture of peptide and cells, indicating promising cell biocompatibility at the modified sample surface. At last, antifouling performance and electrochemical corrosion were tested. Results show that the adhesion rates of Vibrio natriegens and Phaeodactylum tricornutum on the antifouling surface were reduced by 99.85% and 67.93%, respectively from those of untreated samples. Therefore, the modified samples retained superior corrosion resistance. The study provide a simply and green way against biofouling on ship hulls and marine equipment. [ABSTRACT FROM AUTHOR]

Vancomycin (VCM)-induced nephrotoxicity impedes its treatment applications. Thus, it is important to clarify the relevant mechanism. This study investigated phosphoprotein changes attributable to the VCM nephrotoxicity mechanisms. Biochemical, pathological and phosphoproteomic analyses based on C57BL/6 mice were performed to explore the mechanisms.VCM-treated mice showed increased levels of blood urea nitrogen and creatinine, and signs of acute tubular necrotic lesions. Phosphoproteomic profiling identified 3025 differentially phosphorylated phosphopeptides between the model and control group. Gene Ontology enrichment analysis demonstrated that Molecular Function "oxidoreductase activity" and Cellular Component "peroxisome" were markedly enriched. KEGG pathway analysis identified an enrichment in peroxisome pathway and PPAR (peroxisome proliferator-activated receptor) signaling pathways. Parallel reaction monitoring analysis revealed a significant downregulation of CAT, SOD-1, AGPS, DHRS4, and EHHADH at phosphorylation level by VCM. Notably, the phosphorylation of ACO, AMACR, and SCPX was downregulated by VCM, which are the fatty acid β-oxidation-related proteins involved in PPAR signaling pathways. The phosphorylated PEX5 involved in peroxisome biogenesis was upregulated by VCM. Collectively, these findings indicated that VCM-induced nephrotoxicity is closely associated with peroxisome pathway and PPAR signaling pathways. The current study provides important insight into the mechanisms of VCM nephrotoxicity and will aid in the development of preventive and therapeutic strategies against this nephropathy. [ABSTRACT FROM AUTHOR]

Simple Summary: The biological relevance of peptides that originate from non-canonical translational initiation sites have been increasingly recognized over the years. Peptides encoded by open reading frames upstream of canonical protein coding sequences are frequently translated and act as translational regulators, contribute to the immunopeptidome as cellular antigens, and are implicated in various cellular functions through peptide–protein interactions or as part of protein complexes. In this review, we first give an overview of the most relevant technical advances in non-canonical peptide detection. In the second part of the review, we focus on the functional implications of uPeptides and delineate how this largely unexplored compartment of the human peptidome affects tumor biology and may offer new opportunities for targeted and immunological cancer therapy. Recent technological advances have facilitated the detection of numerous non-canonical human peptides derived from regulatory regions of mRNAs, long non-coding RNAs, and other cryptic transcripts. In this review, we first give an overview of the classification of these novel peptides and summarize recent improvements in their annotation and detection by ribosome profiling, mass spectrometry, and individual experimental analysis. A large fraction of the novel peptides originates from translation at upstream open reading frames (uORFs) that are located within the transcript leader sequence of regular mRNA. In humans, uORF-encoded peptides (uPeptides) have been detected in both healthy and malignantly transformed cells and emerge as important regulators in cellular and immunological pathways. In the second part of the review, we focus on various functional implications of uPeptides. As uPeptides frequently act at the transition of translational regulation and individual peptide function, we describe the mechanistic modes of translational regulation through ribosome stalling, the involvement in cellular programs through protein interaction and complex formation, and their role within the human leukocyte antigen (HLA)-associated immunopeptidome as HLA uLigands. We delineate how malignant transformation may lead to the formation of novel uORFs, uPeptides, or HLA uLigands and explain their potential implication in tumor biology. Ultimately, we speculate on a potential use of uPeptides as peptide drugs and discuss how uPeptides and HLA uLigands may facilitate translational inhibition of oncogenic protein messages and immunotherapeutic approaches in cancer therapy. [ABSTRACT FROM AUTHOR]

Legumes are a staple of diets all around the world. In some least developed countries, they are the primary source of protein; however, their beneficial properties go beyond their nutritional value. Recent research has shown that legumes have bioactive compounds like peptides, polyphenols and saponins, which exhibit antioxidant, antihypertensive, anti-inflammatory and other biological activities. Thus, these compounds could be an alternative treatment for inflammatory diseases, in particular, chronic inflammation such as arthritis, obesity and cancer. Nowadays, there is a growing interest in alternative therapies derived from natural products; accordingly, the present review has compiled the bioactive compounds found in legumes that have demonstrated an anti-inflammatory effect in non-clinical studies. [ABSTRACT FROM AUTHOR]

Small ubiquitin-like modifier (SUMO) proteins can reversibly attach covalently or non-covalently to lysine residues of various substrates. The processes are named SUMOylation and de-SUMOylation, which maintain a dynamic balance in the physiological state, and are regulated by SUMO components. However, the dysregulation of components disturbs the balance and alters the functions of target proteins, which causes the occurrence of diseases. To date, certain SUMO components, including SUMO-1, SUMO-2/3, SAE1/Uba2, Ubc9, PIASs (protein inhibitors of activated signal transducer and activator of transcription) and SENPs (SUMO-specific proteases), have been found to participate in the pathogenesis of RA and their potential value as therapeutic targets also have been highlighted. In addition, single nucleotide polymorphisms (SNPs) in the SUMO components have been reported to be associated with disease susceptibility. Until now, only the SNP site of SUMO-4 has been reported in RA. Here we provided a systematic overview of the general characteristics of SUMO components and highlighted a summary of their impact on RA. [ABSTRACT FROM AUTHOR]

Ion transport across biomembranes plays a major role in living cells. This fundamental function is normally carried out by molecules with both a hydrophobic and a hydrophilic side (amphiphilic molecules), which aggregate within the membrane forming a hydrophilic pore (the ion channel) permitting the selective translocation of permeant ions. Countless papers report the conformation of these ion channels in lipid vesicles using several techniques, such as circular dichroism and solid‐state NMR spectroscopies. However, the functional activity of ion channels can only be investigated by varying the transmembrane potential. This is also the situation in which ion channels operate in commercialized drugs with intracellular targeting activities, of great interest in pharmaceutical research. A suitable biomimetic membrane must consist of a conducting or semiconducting support, whose "heart" is a lipid bilayer in contact with the aqueous solution of interest on one side. The other side must comprise a hydrophilic region thick enough to completely decouple the lipid bilayer from the support, giving rise to a "tethered bilayer lipid membrane" (tBLM). This review aims to describe the numerous efforts made over time to approach this goal, the most recent achievements, and the perspectives of future development. Special emphasis will be placed on the electrochemical aspects of tBLMs, and a qualitative overview of the main optical and scanning probe techniques employed will be provided. [ABSTRACT FROM AUTHOR]

Aim: To examine whether functional gene polymorphisms of toll‐like receptor (TLR)1, TLR2, and TLR6 are related to the salivary concentrations of human beta‐defensins (hBDs)‐1, ‐2, ‐3, and human neutrophilic peptide (HNP)‐1. Materials and Methods: Polymorphisms of TLR1 (rs5743618), TLR2 (rs5743708), and TLR6 (rs5743810) were genotyped by PCR‐based pyrosequencing from the salivary samples of 230 adults. Salivary hBD‐1, ‐2, ‐3, and HNP‐1 concentrations were measured using enzyme‐linked immunosorbent assay. General and periodontal health examinations, including panoramic radiography, were available for all participants. Results: The genotype frequencies for wild types and variant types were as follows: 66.5% and 33.5% for TLR1, 95.5% and 4.5% for TLR2, and 25.1% and 74.9% for TLR6, respectively. The TLR2 heterozygote variant group exhibited higher salivary hBD‐2 concentrations than the TLR2 wild‐type group (p =.038). On the contrary, elevated hBD‐2 concentrations were detected in the TLR6 wild‐type group compared with the TLR6 heterozygote and homozygote variant group (p =.028). The associations between TLR6 genotypes and salivary hBD‐2 concentrations remained significant after adjusting them for periodontal status, age, and smoking. Conclusion: hBD‐2 concentrations in saliva are related to TLR2 and TLR6 polymorphisms, but only the TLR6 genotype seems to exhibit an independent association with the salivary hBD‐2 concentrations. [ABSTRACT FROM AUTHOR]

Simple Summary: Immunosurveillance of cancer is mediated by T cell-based recognition of tumor-associated antigens, i.e., short peptides that are presented on the surface of cells on human leukocyte antigen (HLA) molecules. This encourages the analysis of the entirety of HLA-presented peptides, the so-called immunopeptidome, of malignant and benign cells, in order to identify novel therapeutic targets presented exclusively on malignant cells. In the present study, we aim to investigate the role of previously described immunopeptidome-defined antigen presentation in chronic lymphocytic leukemia (CLL) patients for clinical characteristics and disease outcome. We observed that higher yields of presented total and CLL-exclusive peptides were associated with a more favorable disease course, suggesting efficient immunosurveillance in a subgroup of patients and the possibility of further investigating T cell-based therapeutic approaches for CLL. Chronic lymphocytic leukemia (CLL) is characterized by recurrent relapses and resistance to treatment, even with novel therapeutic approaches. Despite being considered as a disease with low mutational burden and thus poor immunogenic, CLL seems to retain the ability of eliciting specific T cell activation. Accordingly, we recently found non-mutated tumor-associated antigens to play a central role in CLL immunosurveillance. Here, we investigated the association of total and CLL-exclusive HLA class I and HLA class II peptide presentation in the mass spectrometry-defined immunopeptidome of leukemic cells with clinical features and disease outcome of 57 CLL patients. Patients whose CLL cells present a more diverse immunopeptidome experienced fewer relapses. During the follow-up phase of up to 10 years, patients with an HLA class I-restricted presentation of high numbers of total and CLL-exclusive peptides on their malignant cells showed a more favorable disease course with a prolonged progression-free survival (PFS). Overall, our results suggest the existence of an efficient T cell-based immunosurveillance mediated by CLL-associated tumor antigens, supporting ongoing efforts in developing T cell-based immunotherapeutic strategies for CLL. [ABSTRACT FROM AUTHOR]

A synthetic peptide 'TDM-623' is a promising hemostatic material for endoscopic surgery in the gastrointestinal tract. However, its stability under gastric/pancreatic secretion has yet to be shown. Hence, this study verified the stability of TDM-623 under digestive enzyme. TDM-623 (1 mL) was applied to a dish and left at 37 °C for gel formation. Artificial gastric/pancreatic juice and saline were applied to each dish (n = 10). The stability of TDM-623 was evaluated at 0 min-48 h. The evaluation methods were the peptide amount dissolved in the solvent measured by high-performance liquid chromatography and morphological changes revealed by photography and eosin staining, as well as physical changes found by weight. The amount of dissolved peptides originating from the TDM-623 gel after gastric juice exposure was negligible compared to its original amount: a maximum of 10.3% dissolved after 48 h. No significant amounts of dissolved peptides were seen at any timepoint of exposure to pancreatic juice. Also, there were neither morphological nor physical changes after exposure to digestive enzymes for 48 h. This study suggests that TDM-623 is sufficiently stable under digestive enzyme. TDM-623 is thus expected to be a durable hemostatic material that protect wound left by endoscopic surgery. [ABSTRACT FROM AUTHOR]

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The role of cardiac biomarkers in diagnosing acute myocardial infarction is undoubted. In the 2020 guidelines of the European Society of Cardiology, the measurement of cardiac peptides to gain prognostic information has a class IIa indication in all patients with ACS. In emergency care, ruling out a non-ST elevation myocardial infarction requires documentation of normal levels of cardiac biomarkers, which remain stable or have very small variations within several hours. This review aims to summarize the current knowledge and recent progresses in the field of cardiac biomarker discovery, from their routine use in emergency rooms to their prognostic roles in modern risk assessment tools. Integrated approaches combining cardiac troponin with other biomarkers of ventricular dysfunction or inflammation, or with modern cardiac imaging in emergency care are also presented, as well as the role of modern algorithms for serial troponin measurement in the modern management of emergency departments. [ABSTRACT FROM AUTHOR]

Background. Lung cancer is a common clinical thoracic malignant tumor, which had a serious impact on the safety of patients, currently ranking first in all malignant tumors in morbidity and mortality, with generally less than 5% survival rate in 5 years. Objective. To investigate the relationship and significance between carcinoembryonic antigen (CEA) and precursor gastrin-releasing peptide (ProGRP) changing levels in bronchoalveolar lavage fluid (BALF) and CT imaging features in patients with peripheral lung cancer. Methods. A total of 90 patients diagnosed with peripheral lung cancer as the lung cancer group and 60 patients with benign lung diseases as the control group in our hospital from May 2019 to October 2021 were selected to compare the differences of CEA and ProGRP in BALF by the classification of CT features. Results. The levels of CEA and ProGRP in the BALF of the lung cancer group were significantly higher than those of the control group; the proportion of patients with lobulation sign, burr sign, ground glass sign, pleural effusion, and lesion diameter ≥ 3.0 cm in the lung cancer group was higher than that in the control group; the CEA level in BALF of lung cancer patients with spicule sign, pleural effusion, and lesion diameter ≥ 3.0 cm was significantly higher than that without these symptoms, while ProGRP level in the BALF of lung cancer patients with lobulation sign, burr sign, ground glass sign, pleural effusion, and lesion diameter ≥ 3. 0 cm was significantly higher than that of lung cancer patients without these symptoms. Conclusion. The check of CEA and ProGRP in BALF in combination with CT features has a certain clinical value for the diagnosis of lung cancer. At the same time, the increased levels of CEA and ProGRP in BALF have a certain correlation with the changes of malignant signs of lung cancer in CT examination. [ABSTRACT FROM AUTHOR]

Aim: Psoriasis is thought to result in vascular diseases, atherogenesis, peripheral insulin resistance, and cardiac comorbidities by causing metabolic function disorders, hypertension, and type 2 diabetes. To our best knowledge, there has not yet been a study evaluating psoriasis patients with regard to copeptin. The present study assesses cardiovascular disease in psoriasis patients based on metabolic function and copeptin levels. Methods: The presented case-control study, which is a type of analytical observational study, included 45 psoriasis patients and an age-sex matched control group admitted to University of Health Sciences Turkey, Istanbul Haseki Training and Research Hospital, Clinic of Dermatology, between March 2016 and May 2017. Patients' blood pressure, height, weight, body mass index, and waistline were measured for both groups. All subjects were also given complete blood count, fasting blood glucose (FBG), uric acid, lipid profile, insulin, C-reactive protein (CRP), copeptin levels, neutrophil-lymphocyte rate, and thrombocyte-lymphocyte rate were analysed. Copeptin was measured by an ELISA kit. Results: Insulin and CRP averages were statistically significantly higher in psoriasis patients than in the control group (p=0.001 for both). The neutrophil-lymphocyte and thrombocyte-lymphocyte rates were significantly higher in psoriasis patients (p=0.008). Insulin resistance was also higher in psoriasis patients (p=0.001). In both the patient and control groups, there was no statistically significant relationship found between copeptin level, general characteristics, and laboratory parameters. Conclusion: To our best knowledge, this is the first study evaluating psoriasis patients with regard to copeptin. Psoriasis patients should be followed up with easily accessible parameters such as neutrophil-lymphocyte rate, thrombocyte-lymphocyte rate, insulin levels, FBG, and uric acid levels. [ABSTRACT FROM AUTHOR]

This communication introduces the first-time application of high-resolution continuum-source molecular absorption spectrometry (HR CS MAS) for the quantification of a peptide. The graphite furnace technique was employed and the tripeptide glutathione (GSH) served as a model compound. Based on measuring sulfur in terms of carbon monosulfide (CS), a method was elaborated to analyze aqueous solutions of GSH. The most prominent wavelength of the CS molecule occurred at 258.0560 nm and was adduced for monitoring. The methodological development covered the optimization of the pyrolysis and vaporization temperatures. These were found optimally to be 250 °C and 2250 °C, respectively. Moreover, the effect of modifiers (zirconium, calcium, magnesium, palladium) on the absorption signals was investigated. The best results were obtained after permanent coating of the graphite tube with zirconium (total amount of 400 μg) and adding a combination of palladium (10 µL, 10 g L−1) and calcium (2 µL, 1 g L−1) as a chemical modifier to the probes (10 µL). Aqueous standard samples of GSH were used for the calibration. It showed a linear range of 2.5–100 µg mL−1 sulfur contained in GSH with a correlation coefficient R2 > 0.997. The developed method exhibited a limit of detection (LOD) and quantification (LOQ) of 2.1 µg mL−1 and 4.3 µg mL−1 sulfur, respectively. The characteristic mass accounted for 5.9 ng sulfur. The method confirmed the general suitability of MAS for the analysis of an oligopeptide. Thus, this study serves as groundwork for further development in order to extend the application of classical atomic absorption spectrometry (AAS). [ABSTRACT FROM AUTHOR]

Traditional database search methods for the analysis of bottom-up proteomics tandem mass spectrometry (MS/MS) data are limited in their ability to detect peptides with post-translational modifications (PTMs). Recently, "open modification" database search strategies, in which the requirement that the mass of the database peptide closely matches the observed precursor mass is relaxed, have become popular as ways to find a wider variety of types of PTMs. Indeed, in one study, Kong et al. reported that the open modification search tool MSFragger can achieve higher statistical power to detect peptides than a traditional "narrow window" database search. We investigated this claim empirically and, in the process, uncovered a potential general problem with false discovery rate (FDR) control in the machine learning postprocessors Percolator and PeptideProphet. This problem might have contributed to Kong et al. 's report that their empirical results suggest that false discovery (FDR) control in the narrow window setting might generally be compromised. Indeed, reanalyzing the same data while using a more standard form of target-decoy competition-based FDR control, we found that, after accounting for chimeric spectra as well as for the inherent difference in the number of candidates in open and narrow searches, the data does not provide sufficient evidence that FDR control in proteomics MS/MS database search is inherently problematic.

Searching for tandem mass spectrometry proteomics data against a database is a well-established method for assigning peptide sequences to observed spectra but typically cannot identify peptides harboring unexpected post-translational modifications (PTMs). Open modification searching aims to address this problem by allowing a spectrum to match a peptide even if the spectrum's precursor mass differs from the peptide mass. However, expanding the search space in this way can lead to a loss of statistical power to detect peptides. We therefore developed a method, called CONGA (combining open and narrow searches with group-wise analysis), that takes into account results from both types of searches─a traditional "narrow window" search and an open modification search─while carrying out rigorous false discovery rate control. The result is an algorithm that provides the best of both worlds: the ability to detect unexpected PTMs without a concomitant loss of power to detect unmodified peptides.

Early diagnosis of cancer can significantly improve survival of cancer patients; however sensitive and highly specific biomarkers for cancer detection are currently lacking for most cancer types. Nullpeptides are short peptides that are absent from the human proteome. Here, we examined the emergence of nullpeptides during cancer development. We analyzed 3,600,964 somatic mutations across 10,064 whole exome sequencing tumor samples spanning 32 cancer types. We analyze RNA-seq data from primary tumor samples to identify the subset of nullpeptides that emerge in highly expresed genes. We show that nullpeptides, and particularly the subset that is highly recurrent across cancer patients, can be identified in tumor biopsy samples. We find that cancer genes show an excess of nullpeptides and detect nullpeptide hotspots in specific loci of oncogenes and tumor suppressors. We also observe that recurrent nullpeptides are more likely to be found in neoantigens, which have been shown to be effective targets for immunotherapy, suggesting that they can be used to prioritize candidates. Our findings provide evidence for the utility of nullpeptides as cancer detection and therapeutic biomarkers. • Nullpeptides are short sequences absent from healthy proteomes. • Nullpeptides emerge due to cancer mutations and found in RNA-seq data. • Asubset of nullpeptides is observed across cancer patients. • Neoantigens are enriched for recurrent nullpeptides. [ABSTRACT FROM AUTHOR]