BackgroundOsteoarthritis is a chronic musculoskeletal disorder characterized mainly by progressive degradation of the hyaline cartilage. Patients with osteoarthritis often postpone seeking medical help, which results in the diagnosis being made at an advanced stage of cartilage destruction. Sustained efforts are needed to identify specific markers that might help in early diagnosis, monitoring disease progression and in improving therapeutic outcomes. We employed a multipronged proteomic approach, which included multiple fractionation strategies followed by high resolution mass spectrometry analysis to explore the proteome of synovial fluid obtained from osteoarthritis patients. In addition to the total proteome, we also enriched glycoproteins from synovial fluid using lectin affinity chromatography.ResultsWe identified 677 proteins from synovial fluid of patients with osteoarthritis of which 545 proteins have not been previously reported. These novel proteins included ADAM-like decysin 1 (ADAMDEC1), alanyl (membrane) aminopeptidase (ANPEP), CD84, fibulin 1 (FBLN1), matrix remodelling associated 5 (MXRA5), secreted phosphoprotein 2 (SPP2) and spondin 2 (SPON2). We identified 300 proteins using lectin affinity chromatography, including the glycoproteins afamin (AFM), attractin (ATRN), fibrillin 1 (FBN1), transferrin (TF), tissue inhibitor of metalloproteinase 1 (TIMP1) and vasorin (VSN). Gene ontology analysis confirmed that a majority of the identified proteins were extracellular and are mostly involved in cell communication and signaling. We also confirmed the expression of ANPEP, dickkopf WNT signaling pathway inhibitor 3 (DKK3) and osteoglycin (OGN) by multiple reaction monitoring (MRM) analysis of osteoarthritis synovial fluid samples.ConclusionsWe present an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis. We believe that the catalog of proteins generated in this study will further enhance our knowledge regarding the pathophysiology of osteoarthritis and should assist in identifying better biomarkers for early diagnosis.
BackgroundRheumatoid arthritis and osteoarthritis are two common musculoskeletal disorders that affect the joints. Despite high prevalence rates, etiological factors involved in these disorders remain largely unknown. Dissecting the molecular aspects of these disorders will significantly contribute to improving their diagnosis and clinical management. In order to identify proteins that are differentially expressed between these two conditions, a quantitative proteomic profiling of synovial fluid obtained from rheumatoid arthritis and osteoarthritis patients was carried out by using iTRAQ labeling followed by high resolution mass spectrometry analysis.ResultsWe have identified 575 proteins out of which 135 proteins were found to be differentially expressed by ≥3-fold in the synovial fluid of rheumatoid arthritis and osteoarthritis patients. Proteins not previously reported to be associated with rheumatoid arthritis including, coronin-1A (CORO1A), fibrinogen like-2 (FGL2), and macrophage capping protein (CAPG) were found to be upregulated in rheumatoid arthritis. Proteins such as CD5 molecule-like protein (CD5L), soluble scavenger receptor cysteine-rich domain-containing protein (SSC5D), and TTK protein kinase (TTK) were found to be upregulated in the synovial fluid of osteoarthritis patients. We confirmed the upregulation of CAPG in rheumatoid arthritis synovial fluid by multiple reaction monitoring assay as well as by Western blot. Pathway analysis of differentially expressed proteins revealed a significant enrichment of genes involved in glycolytic pathway in rheumatoid arthritis.ConclusionsWe report here the largest identification of proteins from the synovial fluid of rheumatoid arthritis and osteoarthritis patients using a quantitative proteomics approach. The novel proteins identified from our study needs to be explored further for their role in the disease pathogenesis of rheumatoid arthritis and osteoarthritis.Sartaj Ahmad and Raja Sekhar Nirujogi contributed equally to this article.
Insect pest infestation is a major cause of crop loss during storage. However, the use of chemical pesticides results in environmental pollution and may cause allergic reactions when consumed. The inhibition of insect digestive enzymes with potential proteinaceous or non-proteinaceous inhibitors could be an effective alternative approach to control infestations. In the present study, we evaluated the specificity of the inhibitor proteins a-amylase and trypsin in 54 genotypes of wheat (Triticum aestivum) against the digestive amylases of the wheat storage insects Rhyzopertha dominica and Tenebrio molitor, human salivary amylase, porcine pancreatic amylase and bovine trypsin. We found that these proteinaceous inhibitors vary in their expression between genotypes, and identified six genotypes in which inhibitor activity was high against the insects but low against human salivary and pancreatic amylase. These findings will significantly aid the selection of wheat genotypes with potential resistance against storage insects and minimum health concerns for humans.
Introduction: Endometrial specimen for abnormal uterine bleeding (AUB) is the one of the commonest specimens received in histopathology laboratory. Histopathological characteristics of endometrial tissues, as assessed by light microscopy, remains the diagnostic standard for the management of AUB. The objective of study is to find out the histopathological pattern of endometrium in AUB in the light of clinical details. Methods: This was a prospective observational study carried out in the department of Pathology, Lumbini Medical College Teaching Hospital for a period of two years from June 2014 to May 2016. Formalin fixed endometrial specimens were processed, paraffin embedded, sectioned at 3-4 µm, stained with hematoxylin and eosin, and studied under light microscopy along with their demographics. Data were collected, entered and analyzed using SPSS version 20. Results: The study included 100 cases of endometrial biopsy specimens with clinical diagnosis of AUB. Menstrual disturbances was found in wide age range between 17-75 years with the mean age of 45 (SD=13.36) years. Menorrhagia was the commonest (n=60, 60%) clinical presentation. Most (n=85; 85 %) endometrium had non-neoplastic lesions. Among them, normal endometrial patterns were commonest (n=50, 50 %). Neoplastic lesions (n=15, 15%) were distributed in all menstruation status with majority in postmenopause (n=7, 7%) and included malignant cases (n=5, 5%) among others. Conclusion: Post-menopausal bleeding was common presentation among women with malignant and premalignant disease which was present in 15% of the cases together. Timely evaluation of AUB by histopathology can be life saving with early tissue diagnosis and management.
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