Bromelain is a mixture of proteolytic enzymes derived from pineapple (Ananas comosus) fruit and stem possessing several beneficial properties, particularly anti-inflammatory activity. However, the molecular mechanisms underlying the anti-inflammatory effects of bromelain are unclear. This study investigated the anti-inflammatory effects and inhibitory molecular mechanisms of crude and purified rhizome bromelains on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. RAW264.7 cells were pre-treated with various concentrations of crude bromelain (CB) or purified bromelain (PB), and then treated with LPS. The production levels of pro-inflammatory cytokines and mediators, including nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)- were determined by Griess and ELISA assays. The expressions of inducible nitric oxide synthetase (iNOS), cyclooxygenase (COX)-2, nuclear factor kappa B (NF-B), and mitogen-activated protein kinases (MAPKs)-signaling pathway-related proteins were examined by western blot analysis. The pre-treatment of bromelain dose-dependently reduced LPS-induced pro-inflammatory cytokines and mediators, which correlated with downregulation of iNOS and COX-2 expressions. The inhibitory potency of PB was stronger than that of CB. PB also suppressed phosphorylated NF-B (p65), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha, extracellular signal-regulated kinases, c-Jun amino-terminal kinases, and p38 proteins in LPS-treated cells. PB then exhibited potent anti-inflammatory effects on LPS-induced inflammatory responses in RAW264.7 cells by inhibiting the NF-B and MAPKs-signaling pathways.

Background: Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that affecting multi-organs injury and failure. The rapid, precision, and specificity prognosis by laboratory investigations could prevent active stage and severity in the disease. Aims: To systematically explore and investigate the candidate serum protein for development into the novel biomarker for severity prognosis of SLE patients. Methods: The proteins previously reported in abnormal level in serum/plasma of SLE patients since 2014-2020 were comprehensive collected. Thereafter, these serum proteins that found in other diseases were excluded. The association with molecules related to SLE severity of these candidate proteins were then predicted using bioinformatics STIRNG tool. The level of altered protein, which had the strong interaction to the severity molecules in serum of SLE patients was validated by Western blot, analyzed the correlation with anti-nuclear antigen (ANA) and performance of diagnosis, respectively. Results: From 26 collected serum/plasma proteins, alpha-1-antitrypsin protein was found the abnormal level in only SLE patients and strongly associated with severity molecules including C-reactive protein (CRP), complement C3, and C4. Additionally, the validation of serum alpha-1-antitrypsin in SLE patients exhibited the higher level than healthy controls and also had the positive correlation with ANA titer (r = 0.710). Furthermore, the area under ROC curve for diagnostic power of alpha-1-antitrypsin was 0.970 with 100% sensitivity and 90% specificity at cut-off 0.131/total serum protein. Conclusions. The higher level of alpha-1-antitrypsin in serum samples of SLE patients indicated as the novel biomarker for reliable and specific prognosis of disease severity.

Introduction: Antidesma thwaitesianum Müll. Arg is a tropical fruit, which has been commonly used for healthy food and traditional herbal medicine. This study aimed to investigate the anti-inflammatory effects of A. thwaitesianum fruit extract (AFE) rich in 5-hydroxymethylfurfural (5-HMF) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Methods: The chemical composition of AFE was analysed by gas chromatography/mass spectrometry (GC/MS). RAW264.7 cells were used as an in vitro inflammatory response model. RAW264.7 cells were pre-treated with various concentrations of AFE or 5-HMF and subsequently treated with LPS. Cell viability was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The production levels of pro-inflammatory cytokines and mediators, including nitric oxide (NO), interleukin (IL)-1β, and tumour necrosis factor (TNF)-α were determined by the Griess assay and enzyme-linked immunosorbent assay. The protein expression of inducible nitric oxide synthase (iNOS) was examined by western blot analysis. Results: AFE had a high content of 5-HMF (61.03% ± 0.49%). Pre-treatment with AFE and 5-HMF markedly reduced LPS-induced pro-inflammatory mediators and cytokines, namely NO, IL-1β, and TNF-α, in RAW264.7 cells; this reduction correlated with downregulation of iNOS expression. Conclusion: This study suggests that A. thwaitesianum fruit extract containing 5-HMF could modulate the LPS-induced inflammatory response by inhibiting NO, IL-1β, and TNF-α production and iNOS expression. A. thwaitesianum fruit extract rich in 5-HMF could be considered a potential therapeutic agent for the prevention of inflammation.

The severity and spread of tuberculosis, a major burden, can be prevented by more rapid and accurate laboratory diagnosis. The purpose of this study is to systematically explore candidate serum proteins in patients with Mycobacterium tuberculosis infection for further application as novel biomarkers. Our study was performed in two major steps: screening of the literature for potential biomarkers, and then validation of their levels in patients and controls. Many serum/plasma proteins previously reported to be abnormally expressed in patients with tuberculosis between 2012 and 2021 were comprehensively assembled. The biological role in tuberculosis was also predicted for each using the bioinformatics tool STRING. Candidate proteins found to have the same expression in other related diseases were excluded. Subsequently, the serum level of the candidate serum/plasma protein that met the aforementioned criteria was validated by sandwich ELISA; diagnostic performance was analysed by the area under the curve (AUC) of the receiver operating characteristic (ROC). From 103 collected serum/plasma proteins, coronin-1A was found to have abnormal expression only in patients with tuberculosis and was associated with tuberculosis. In addition, the validation of coronin-1A in the serum of patients with pulmonary tuberculosis revealed a higher level than in that of healthy individuals. Furthermore, the area under the ROC curve for diagnostic power of coronin-1A was 0.866, with high sensitivity and specificity at a cut-point of approximately 52.7 ng/mL. We concluded that the level of serum coronin-1A might serve as a novel biomarker for alternative laboratory examination to effectively distinguish patients with tuberculosis from those with other related diseases and healthy individuals.

Fibronectin, an extracellular matrix (ECM) protein, has been thought to be involved in pathogenic mechanisms of kidney stone disease, especially calcium oxalate (CaOx) type. Nevertheless, its precise roles in modulation of CaOx crystal remained unclear. We thus performed a systematic evaluation of effects of fibronectin on CaOx monohydrate (COM) crystal (the major causative chemical crystal in kidney stone formation) in various stages of kidney stone pathogenesis, including crystallization, crystal growth, aggregation, adhesion onto renal tubular cells, and invasion through ECM in renal interstitium. The data showed that fibronectin significantly decreased crystallization, growth and adhesive capability of COM crystals in a dose‐dependent manner. In contrast, COM crystal aggregation and invasion through ECM migration chamber were significantly enhanced by fibronectin in a dose‐dependent fashion. Sequence analysis revealed two and six calcium‐binding and oxalate‐binding domains, respectively. Saturation of fibronectin with oxalate prior to crystal modulation assays could abolish all the effects of fibronectin, whereas saturation with calcium had minimal interference. These data strongly indicate the dual functions of fibronectin, which serves as an inhibitor for COM crystallization, crystal growth and adhesion onto renal tubular cells, but on the other hand, acts as a promoter for COM crystal aggregation and invasion through ECM. Moreover, its crystal modulatory activities are mainly due to oxalate‐binding capability. Support or Funding Information This study was supported by Mahidol University research grant and the Thailand Research Fund (IRN60W0004 and IRG5980006). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

In the original publication, the Western blot images presented in Figures 3C, 5A and 6A contained errors in image presentation