Concentrations Of Cathepsin Research Articles

The SPRi determination of cathepsin L and S in plasma and peritoneal fluid of women with endometriosis

PurposeEndometriosis is a common disease with a complex pathomechanism and atypical symptoms, often leading to delayed diagnosis. Currently, the sole method for confirming the presence of the disease is through laparoscopy and histopathological examination of collected tissue. However, this invasive procedure carries potential risk and complications, necessitating the exploration of non-surgical diagnostic methods for endometriosis. This study aims to analyze peritoneal fluid and plasma samples for the expression of cathepsin L and cathepsin S to identify potential biomarkers for non-invasive diagnostic approaches to endometriosis. Material and methodsIn this cross-sectional study, plasma and peritoneal fluid samples were obtained during laparoscopy from 63 patients diagnosed with chronic pelvic pain or infertility. The study group consisted of women with confirmed endometriosis. The concentrations of cathepsins L and S were determined using an SPRi biosensor. ResultsThe study did not reveal significant differences in the concentrations of cathepsin L and cathepsin S between the control group and the study group, both in peritoneal fluid and plasma. ConclusionsBased on the results of this study, it appears that cathepsins L and S are not suitable candidates as biomarkers for endometriosis.

Differential patterns of lysosomal dysfunction are seen in the clinicopathological forms of primary progressive aphasia.

Increasing evidence implicates endo-lysosomal dysfunction in frontotemporal dementia (FTD). 18 proteins were quantified using a mass spectrometry assay panel in the cerebrospinal fluid of 36 people with the language variant of FTD, primary progressive aphasia (PPA) (including 13 with non-fluent variant (nfvPPA), 11 with semantic variant (svPPA), and 12 with logopenic variant (lvPPA)) and 19 healthy controls. The concentrations of the cathepsins (B, D, F, L1, and Z) as well as AP-2 complex subunit beta, ganglioside GM2 activator, beta-hexosaminidase subunit beta, tissue alpha L-fucosidase, and ubiquitin were decreased in nfvPPA compared with controls. In contrast, the concentrations of amyloid beta A4 protein, cathepsin Z, and dipeptidyl peptidase 2 were decreased in svPPA compared with controls. No proteins were abnormal in lvPPA. These results indicate a differential alteration of lysosomal proteins in the PPA variants, suggesting those with non-Alzheimer's pathologies are more likely to show abnormal lysosomal function.

Harnessing protein sensing ability of electrochemical biosensors via a controlled peptide receptor–electrode interface

BackgroundCathepsin B, a cysteine protease, is considered a potential biomarker for early diagnosis of cancer and inflammatory bowel diseases. Therefore, more feasible and effective diagnostic method may be beneficial for monitoring of cancer or related diseases.ResultsA phage-display library was biopanned against biotinylated cathepsin B to identify a high-affinity peptide with the sequence WDMWPSMDWKAE. The identified peptide-displaying phage clones and phage-free synthetic peptides were characterized using enzyme-linked immunosorbent assays (ELISAs) and electrochemical analyses (impedance spectroscopy, cyclic voltammetry, and square wave voltammetry). Feasibilities of phage-on-a-sensor, peptide-on-a-sensor, and peptide-on-a-AuNPs/MXene sensor were evaluated. The limit of detection and binding affinity values of the peptide-on-a-AuNPs/MXene sensor interface were two to four times lower than those of the two other sensors, indicating that the peptide-on-a-AuNPs/MXene sensor is more specific for cathepsin B (good recovery (86–102%) and %RSD (< 11%) with clinical samples, and can distinguish different stages of Crohn’s disease. Furthermore, the concentration of cathepsin B measured by our sensor showed a good correlation with those estimated by the commercially available ELISA kit.ConclusionIn summary, screening and rational design of high-affinity peptides specific to cathepsin B for developing peptide-based electrochemical biosensors is reported for the first time. This study could promote the development of alternative antibody-free detection methods for clinical assays to test inflammatory bowel disease and other diseases.

Cathepsin L involved in the freezing resistance of murine normal hatching embryos and dormant embryos

The cryopreservation of mammalian embryos is an important technology in embryo engineering. The discovery and application of the embryo's own high freezing resistance factors are the main methods to improve the utilization of mammalian embryos in cryopreservation. Cathepsin L gene expression in the frozen and thawed dormant embryos displayed a significant difference from those normal hatched ones. The aim of the present study was to dig out the potential role of Cathepsin L in anti-freezing capacity of murine blastocysts by investigating the location and expression of Cathepsin L in frozen and thawed both activated and dormant hatching blastocysts. Different concentrations of Cathepsin L recombinant protein and E-64d were then respectively added into the embryo cryoprotectant and pre-cryo culture medium. Our results found that down-regulation of Cathepsin L improves the freezing resistance of murine normal hatching embryos by reducing apoptosis. Cathepsin L inhibitors can be used to improve the efficiency of cryopreservation and recovery of blastocysts in vitro. Our study provides a theoretical basis for the further development and application of Cathepsin L.

Bioinformatical analysis of the level of osteomarkers in the gingival fluid in systemic osteoporosis and local osteo-destructive processes in the dentoalveolar region

The informational value of assessing biochemical markers in the gingival fluid of patients with various systemic and local changes in bone tissue was analyzed in a sample of 57 patients and 15 healthy people. The patients were differentiated into 3 groups: 1) 20 patients with ulcerative colitis and steroid osteoporosis due to glucocorticoid therapy; 2) 22 patients diagnosed with cancer of the mucous membrane of the floor of the mouth with erosions of the cortical bone or dental socket by a primary tumor; 3) 15 patients with moderate dental peri-implantitis. Gingival fluid was used as a test material. The content of cathepsin K, tartrate-resistant acid phosphatase, and bone fraction of alkaline phosphatase was determined. In steroidal osteoporosis, the cathepsin K level in the gingival fluid was found to be statistically significantly increased compared to other markers under study. In group 2, with tumor osteodestruction in the gingival fluid, a 10-fold increase in the concentration of cathepsin K and tartrate-resistant acid phosphatase was observed against the background of a moderate increase in the bone fraction of alkaline phosphatase content. In osteodestruction associated with inflammatory dentoalveolar processes, a moderate increase in the concentration of cathepsin K and tartrate-resistant acid phosphatase was observed. Thus, the determination of osteomarkers in the gingival fluid can be considered informative in terms of distinguishing between systemic and local changes in the structure of bone tissue.

Urinary Levels of Cathepsin B in Preterm Newborns.

Increased investment in perinatal health in developing countries has improved the survival of preterm newborns, but their significant multiorgan immaturity is associated with short and long-term adverse consequences. Cathepsin B, as a protease with angiogenic properties, may be related to the process of nephrogenesis. A total of 88 neonates (60 premature children, 28 healthy term children) were included in this prospective study. We collected urine samples on the first or second day of life. In order to determine the concentration of cathepsin B in the urine, the commercially available enzyme immunoassay was used. The urinary concentrations of cathepsin B normalized with the urinary concentrations of creatinine (cathepsin B/Cr.) in newborns born at 30–34, 35–36, and 37–41 (the control group) weeks of pregnancy were (median, Q1–Q3) 4.00 (2.82–5.12), 3.07 (1.95–3.90), and 2.51 (2.00–3.48) ng/mg Cr, respectively. Statistically significant differences were found between the group of newborns born at 30–34 weeks of pregnancy and the control group (p < 0.01), and between early and late preterm babies (PTB) (p < 0.05). The group of children born at 35–36 weeks of pregnancy and the control group did not differ significantly. This result suggests that the elevated urinary cathepsin B/Cr. level may be the result of the kidneys’ immaturity in preterm newborns.

Laboratory control of the osteoresorptive effects of glucorcorticoids in patients with purpose colitis.

The aim of the work was to develop an informative method for laboratory monitoring of osteoresorbent action during systemic administration of glucocorticoids in patients with ulcerative colitis. The study included 54 patients with ulcerative colitis aged 18 to 44 years: 35 (64,8%) men and 19 (35,2%) women. In patients of the clinical group before and after the first, second, third courses of glucocorticosteroids, as well as during the formation of steroid dependence, the concentration of the osteoresorption marker cathepsin K was determined simultaneously in the blood serum and gingival fluid by enzyme immunoassay. The concentration of the osteomarker was compared with the parameters of the densitometric density of the lumbar vertebrae L1-L4 during X-ray examination. It was found that with the systemic use of glucocorticoids in patients with ulcerative colitis, the concentration of cathepsin K in the gingival fluid increased earlier than in the blood serum. It was found that with an increase in the concentration of cathepsin K in the gingival fluid of more than 2,6 pmol/l in conditions of systemic administration of glucocorticosteroids, the risk of osteoporosis increased with a diagnostic sensitivity of 81,8% and a specificity of 74,4% (p=0,0001).The diagnostic accuracy was 78,1%. With an increase in the concentration of cathepsin K in the gingival fluid above the differential separation level (2,6 pmol/l), the risk of developing osteoporosis increased 3,2 times (p= 0,0001). The study developed a methodological and informational algorithm has been developed for the non-invasive control of steroidal osteoporosis in patients with ulcerative colitis with systemic use of glucocorticoids by assessing the concentration of cathepsin K in the gingival fluid.

Diagnostic possibilities of laboratory assessment of cathepsin K activity in gingival and peri-implant fluid under normal conditions and periimplantitis

The aim of the work was to develop a non-invasive laboratory method for local control of the severity of osteo-destructive processes in the bone tissue of the jaws after dental implantation. The study comprised 75 individuals aged 18 to 65. The main group included 40 patients with a diagnosis of dental periimplantitis (ICD K 10.2). A comparison group (n=20) included patients after dental implantation without postoperative complications. The control group (n=25) consisted of individuals without dental pathology. Cathepsin K (CTSK) (Human) ELISA Kit (Cloud-Clone Corp., USA) was used to determine the concentration of cathepsin K by the enzyme immunoassay in the gingival or peri-implant fluid. The concentration of cathepsin K in the gingival fluid in the patients of the control group was 1.7±0.3 pmol/l, in the comparison group the concentration of the studied biomarker corresponded to 2.3±0.4 pmol/l. In the main group, the concentration of cathepsin K in the exudate of peri-implantation pockets increased 2.8 times (p<0.05) and corresponded to 4.9±0.5 pmol/l. The concentration of cathepsin K in the gingival fluid normally did not depend on age and gender, which increased the independence of the osteomarker for characterizing the osteoresorbent process in the peri-implantation area. After dental implantation, an excess of cathepsin K concentration above the level of 2.7 pmol/l with a diagnostic sensitivity of 82.5% and a specificity of 83.7% suggests a high risk of developing peri-implantitis. The concentration of cathepsin K may be seen as valuable marker for local inflammatory and destructive processes in the contents of the peri-implantation area.

Diagnostic value of biomarkers of bone metabolism in the gingival fluid in inflammatory-destructive and tumor oral pathology

The aim of the work was to determine the informativeness of the assessment of biochemical markers in the gingival fluid with destructive bone changes in the peri-implantation area, primary tumor lesion of the alveolar processes of the jaws and metastatic destruction of the jaw bone tissue in cancer of the large salivary glands. The study included 51 patients aged 30 to 75 years of both sexes. Depending on the genesis of destruction of the bone tissue of the lower jaw, three groups of patients were identified. Group 1 - 20 patients with peri-implantitis. Group 2 - 22 patients diagnosed with cancer of the oral floor mucosa (ICD C04) with erosion of the cortical bone or dental socket by a primary tumor. Group 3 - 9 patients with adenocarcinoma of the large salivary glands with metastatic lesions of the lower jaw. 15 people were practically healthy persons without periodontal pathology. The test material was gingival fluid or exudate from peri-implantation pockets. The content of cathepsin K, tartrate-resistant acid phosphatase (TAP) was determined by the enzyme immunoassay, and the bone fraction of alkaline phosphatase (BAP) was determined by the kinetic method. As a result, it was found that in group 1 in the exudate of the peri-implantation sulcus, the content of cathepsin K increased in comparison with the control by 2.5 times (p<0.05), TAP by 1.6 times (p<0.05). in groups 1 and 2, there was a moderate increase in the production of activated osteoblasts of KAP. In cancer of the mucous membrane of the floor of the mouth, a tenfold increase in the concentration of cathepsin K and TAP in the gingival fluid was observed. With metastatic lesions of the jaw in patients with adenocarcinoma of the large salivary glands, along with a multiple increase in the concentration of cathepsin K and TAP in the gingival fluid, there was a twenty-fold increase in the bone formation marker BAP, which indicated a mixed type of metastases and was a differential diagnostic criterion for metastatic lesion of bone tissue in comparison with the primary metastatic lesion of bone tissue. Thus, biochemical markers are sensitive indicators of remodeling of bone tissue both in local inflammatory and destructive processes in peri-implantitis and in primary and metastatic tumor lesions of the jaws.

Effectiveness of nicotinamide adenine nucleotide hydride in the complex treatment of patients with peri implantitis

Background: In treating peri-implantitis, dentists pay attention to drugs that do not contain chemically aggressive agents. They rather use those having natural mechanisms for normalization of the functional activity of cells and tissues in addition to local antiseptic and antibacterial agents.&#x0D; The aim of the study was to determine the clinical effectiveness of nicotinamide adenine nucleotide hydride dental gel in the treatment of peri-implantitis.&#x0D; Materials and methods: Depending on the therapeutic tactics patients (n = 40), we divided patients into two groups. In Group 1 (n=20), we used a standard treatment of peri-implantitis including professional hygiene measures, antiseptic treatment and the use of antibacterial agents. In Group 2 (n = 20), the complex treatment included photodynamic effects of 1% photosensitizer gel instead of the antibacterial therapy.&#x0D; Results: Due to the use of the photodynamic therapy in addition to standard treatment of group 2 patients, the concentration of a tartrate-resistant acid phosphatase in the peri-implantation exudate reached the level of healthy people in the gingival fluid. In addition, the concentrations of cathepsin K and interleukin-6 were as close as possible to the control level.&#x0D; Conclusion: The use of NADH for peri-implantitis helps to eliminate inflammatory phenomena and stimulates the regeneration of the oral mucosa.

Information Algorithm for Non-Invasive Evaluation of Osteoresorbable Side Effects of Glucocorticosteroids in Patients with Ulcerative Colitis

A study involving 45 patients with ulcerative colitis found that the activity of the Cathepsin K bone resorption marker measured by an enzyme-linked immunosorbent assay in gingival fluid could be used as a non-invasive control over the development of steroid osteoporosis caused by systemic use of glucocorticoids. When the concentration of Cathepsin K in the gingival fluid exceeded 3.7 pmol/L, the risk of developing osteoporosis in patients with ulcerative colitis taking glucocorticosteroids on a regular basis increased by 7.9 times with a diagnostic accuracy of 82.6% (p=0.002). As a result of the study, an effective method for non-invasive assessment of osteoresorbable side effects of glucocorticosteroids in autoimmune diseases was proposed.

Melphalan-monomethoxypolyethylene glycol-based pH/enzyme double-response polymer prodrug nanoparticles for enhanced drug stability and anticancer efficacy

To improve hydrolytic stability and biological half-life and anticancer efficacy of the anti-cancer drug melphalan (Mel), a novel esterification strategy was developed to fabricate a pH/enzyme dual-responsive polymer prodrug (mPEG-Mel) via the formation of an ion bond between the carboxyl group in Mel and diazotized p-aminobenzoate-terminated poly(ethylene glycol) monomethyl ether (mPEG-N2+) followed by light-induced conversion of the ionic bond to an ester bond. The prodrug could self-assemble into nanoparticles. Under physiological conditions (pH 7.4), the nanoparticles exhibited a regular core-shell structure with about 100 nm, which could effectively inhibit the hydrolysis of pharmacophores in Mel structure and showed the long-circulation ability. While pH dropped to 5.0, an obvious increase in the nanoparticle size was observed and enhanced by different concentrations of cathepsin B (CTB), reaching a diameter of 675 nm or more in the presence of 2.0 µg/mL CTB. The nanoparticles showed minimal drug release at pH 7.4 (less than 10% in 24 h); however, drug release could be significantly accelerated in a CTB-containing medium at pH 5.0. Cell experiments showed that the half-inhibitory concentration (IC50) of nanoparticles was 1.00 µg/mL, which was about 5 times lower than that of free Mel (5.00 µg/mL), and indicating that the nanoparticles were more cytotoxic to cancer cells (4T1 cells). However, significantly reduced cytostatic effects were observed for normal cells (3T3 cells). In addition, cell uptake observation further supported the above experimental results, a stronger red fluorescence in 4T1 cells was observed for NR-loaded nanoparticles, which meant that the nanoparticles could release drug in response to pH/enzyme stimulation. Thus, mPEG-Mel based pH/enzyme double-response polymer prodrug nanoparticles could effectively enhance drug stability and anticancer efficacy.

The Relationship between the Concentration of Cathepsin A, D, and E and the Concentration of Copper and Zinc, and the Size of the Aneurysmal Enlargement in the Wall of the Abdominal Aortic Aneurysm

Despite advances in diagnostics and treatment, aortic aneurysms are an important clinical problem, mainly due to the accompanying complications that may lead to direct loss of life, also the number of diagnosed and operated aneurysms is constantly increasing. The aim of this study is to determine the relationship between the concentration of lysosomal peptidases cathepsin A, D, and E in the wall of the abdominal aortic aneurysm and the concentration of copper and zinc, and the size of the aneurysm widening in the wall of the abdominal aortic aneurysm. The study included 27 patients with abdominal aortic aneurysm from the Department of Vascular Surgery and Transplantation of the University Clinical Hospital in Bialystok. The research material was the wall of the abdominal aortic aneurysm collected intraoperatively. The control material consisted of fragments of the abdominal aorta obtained from organ donors for transplantation. The concentration of cathepsin A, D, and E was determined using enzyme-linked immunosorbent assays. Concentrations of copper and zinc were determined by flame atomic absorption spectrometry after prior mineralization of the samples. All patients were interviewed and asked about basic demographic data, comorbidities, and risk factors for cardiovascular disease to which they were exposed in the past. The statistical analysis was performed using Statistica 10 statistical package. Mann-Whitney U-tests were used and also Spearman's r correlation assuming a significance level of P<0.05. The concentration of cathepsin A, D, and E was higher in the aortic wall altered by the aneurysm than in the wall of the control aorta (P<0.05). The analysis of the data showed that there was a positive correlation between the concentration of cathepsin A and D and the width of the aneurysmal widening (r=0.699 and 0.750, respectively). There was no correlation between cathepsin E concentration and aneurysm width. The higher contents of cathepsin A, D, and E in the wall of the aortic aneurysm than in the normal aortic wall, as well as a positive correlation between the concentration of cathepsin A and D and the width of the aneurysmal widening, allow to assume the participation of these enzymes in the pathogenesis of the aneurysm.

Circulating cathepsin B and D in pregnancy

The purpose of this prospective study was to investigate the changes in the circulating levels of cathepsin B and D in pregnancy. We obtained longitudinal cathepsin B and D levels in 76 healthy pregnant women in the first and third trimesters and compared these levels with 20 non-pregnant controls. The plasma levels of soluble cathepsin B and D were measured using an enzyme-linked immunosorbent assay kit. The cathepsin D concentrations in the third trimester were significantly higher than that in the first trimester (p < .001), and the cathepsin D levels in the first trimester were significantly lower than that in the non-pregnant controls (p = .002). However, there was no significant difference in the cathepsin B level throughout pregnancy compared to the non-pregnant controls. Our study is unique in evaluating the longitudinal changes in the cathepsin B and D levels in pregnancies without obstetric complications. The results implicate that changes in the levels of cathepsins might be essential in placentation. Therefore, molecular and genetic studies on cathepsin B and D are needed to understand the roles of these enzymes in pregnancy, thereby contributing to the understanding of placentation.Impact statementWhat is already known? Matrix metalloproteinases (MMP) have been widely studied, and their function is very important in the normal implantation process. The level of MMP-9 is known to increase throughout pregnancy, while the level of MMP-2 decreases in the first trimester. In addition to MMPs, other proteases are important for placental development; cathepsins B and D are two of the proteases that are involved in the normal placentation process. The function of cathepsin D is related to MMPs because this protease can activate MMPs either directly or indirectly. Nevertheless, the role of circulating cathepsins in pregnancy has not yet been fully elucidated.What do these results add? This study provides evidence, for the first time, that there are fluctuations of plasma cathepsin D level and there are no changes in the plasma cathepsin B level in a normal pregnancy. Moreover, we demonstrated that a cathepsin D level is significantly decreased in the first trimester compared to the non-pregnant controls, and that the level is markedly elevated in the third trimester.What are the implications of these findings for clinical practice and/or further research? Cathepsins B and D should be further studied locally in the placenta to explain the differences in the concentration of cathepsin D and no changes in cathepsin B, thereby exploring their exact roles.

Relationship between bone remodeling markers, bone mineral density, and severity of coronary atherosclerosis in men with stable coronary heart disease

The development of atherosclerosis is a complex multifactorial process, in which the markers of bone formation and resorption play an important role and which is closely related to the calcification of the vessel intima and fibrous plaques. Objective: to assess the relationship between bone remodeling markers (cathepsin K, C-terminal telopeptide of type I collagen (CTX-I) and osteopontin), bone mineral density (BMD), and severity of coronary atherosclerosis in men with stable coronary heart disease (CHD). Patients and methods. The investigation enrolled 102 male patients with verified stable CHD. Coronary angiographic and densitometric findings and blood cathepsin K, osteopontin, and CTX-I concentrations were assessed. Results. The concentration of cathepsin K and CTX-I was significantly higher and that of osteopontin was significantly lower in patients with CHD than in men without CHD. There was no association of the level of bone remodeling markers with the type of coronary artery lesion and the severity of coronary atherosclerosis. Cathepsin K concentrations in patients with a high SYNTAX score of coronary atherosclerosis were shown to be 5.5 times lower in the presence of osteopenic syndrome (OPS) than in those with the similar severity of atherosclerosis and normal BMD. Analysis of osteopontin and CTX-I levels from the standpoint of the presence of OPS suggests that there are no differences in relation to both the type of coronary vessel lesion and its severity. Conclusion. Current data suggest that there are common mechanisms for the development of two socially significant conditions: atherosclerosis and osteoporosis (OP). The phenomenon of concomitant development of these diseases has been studied mainly in postmenopausal women. Today, however, OP is also increasingly found in men, associating with more severe manifestations of coronary atherosclerosis than in patients with no signs of osteopenia.

Inhibitory assay for degradation of collagen IV by cathepsin B with a surface plasmon resonance sensor

We describe a simple method for evaluating the inhibition of collagen IV degradation by cathepsin B with a surface plasmon resonance (SPR) biosensor. The change in the SPR signal decreased with an increase in the concentration of cathepsin B inhibitors. The order of the inhibitory constant (Ki) obtained by the SPR method was CA074Me≈Z-Phe-Phe-FMK < leupeptin. This order was different from that obtained by benzyloxycarbonyl-Phe-Phe-Fluoromethylketone (Z-Phe-Phe-FMK) as a peptide substrate. The comparison of Ki suggested that CA074 and Z-Phe-Phe-FMK inhibited exopeptidase activity, and leupeptin inhibited the endopeptidase activity of cathepsin B more strongly.

Design, Synthesis and In-Silico Studies of Novel Chalcones as Anti-Prostate Cancer and Cathepsin B Inhibitors

The high mortality rate in prostate cancer patients could be mainly attributed to the invasiveness and metastasis of advanced prostate cancer. Cathepsin B, a lysosomal cysteine, that proved to play a significant role in the invasion and metastasis of prostate cancer, therefore, the search for selective cathepsin B inhibitors could be considered an active area of research. Chalcones have proved to be effective anticancer agents against most human malignancies, they have also been reported to exhibit significant cathepsin B inhibition. In the present study, a series of novel N-(4-3-(pheny)-1-prop-2-en-1-one phenyl)benzamide derivatives (5a-p) were synthesized, characterized and evaluated for their anti-prostate cancer activities against PC-3 prostate adenocarcinoma cell line using the SRB method. The cathepsin B inhibition potential was further tested by the using enzyme-linked immunosorbent assay. Moreover, a pharmacophore model was constructed, a QSAR study was carried out where a model was successfully built from which the physicochemical parameters were correlated to the activity. Furthermore, molecular docking studies were also carried out to determine the binding mode of the active compounds into the active site of Cathepsin B enzyme. The furyl derivative 5p was the most active candidate, (IC50 = 5.597 μΜ), against prostate cancer cell line, PC-3. This derivative also demonstrated 50.4% reduction in concentration of cathepsin B, additionally, compounds 5b and 5m showed 53.2 and 56.6% reduction in concentration of cathepsin B, respectively. In conclusion, compounds 5b, 5m and 5p showed good activity both as antiproliferative and as inhibitors of Cathepsin B production. The developed QSAR model was found statistically significant and has good predictive power. Docking studies of the active derivatives maintained the essential key interactions, specially with the cysteine residue, Cys29.

Determination of cathepsin G in endometrial tissue using a surface plasmon resonance imaging biosensor with tailored phosphonic inhibitor

ObjectiveCathepsin G is a serine peptidase whose physiological role is mainly associated with an early immune response, anti-microbial activity as well as platelet activation or hydrolysis of coagulation factors. In addition, since the activity of cathepsin G has been associated with the development of various pathological disorders, the measurement of its activity in patient samples is of high interest. Unfortunately, the usefulness of common immunological methods is limited, since they cannot distinguish between catalytically active and inactive protease. Study designHere we present the application of recently developed Surface Plasmon Resonance-based biosensor for the detection of active cathepsin G in human endometrium samples. The key element of the system is based on the irreversible binding of cathepsin G to its specific phosphonic-type inhibitor immobilized on the surface of the gold chip. The concentration of cathepsin G was measured in tissue samples from the group of patients with endometriosis as well as in the control group. ResultsThe level of cathepsin G ascertained in endometrium tissue samples was over twice as high for the group of patients suffering from endometriosis as compared to the control group, with the median values of 0.5pmol/mg and 0.2pmol/mg, respectively. ConclusionThe SPR sensor armed with a specific irreversible phosphonic inhibitor represents a highly useful tool for the determination of catalytically active cathepsin G concentration in endometrial tissue.

Cathepsin S is increased in cerebrospinal fluid from patients with neuropathic pain—A support of the microglia hypothesis in humans

Abstract Aims Cathepsin S has been reported to be a biomarker of spinal microglial activation, a process suggested to be involved in the pathophysiology of chronic neuropathic pain. So far this has been shown only in animal experiments. The aim of this study was to investigate the concentrations of cathepsin S in human cerebrospinal fluid (CSF) samples from a well-defined patient cohort suffering from neuropathic pain as compared to controls. Methods CSF samples from patients suffering from chronic neuropathic pain (n = 14) were analyzed for cathepsin S levels using commercial sandwich ELISAs (DY1183, R&amp;D Systems, Minneapolis, MN, USA). Control CSF was sampled from patients undergoing minor urological surgical procedures under spinal anaesthesia (n = 70), having no obvious pain suffering. Results The neuropathic pain group had significantly higher levels of CSF cathepsin S (median 15189 pg/mL, range 3213–40,040), than the control group (median 5911 pg/mL, range 1909–17,188) (p &lt; 0.005, Mann–Whitney U-test). Conclusion The results support the existence of microglial activation in chronic neuropathic pain patients. CSF Cathepsin S may serve as a potential biomarker for this specific mechanism linked to neuropathic pain. In the future, Cathepsin S inhibiting drugs might become a new treatment alternative for neurophatic pain.

Sympathetic Nervous System

Activation of the sympathetic nervous system after myocardial infarction results in the mobilization of hematopoietic stem cells, causing an inflammatory boost that accelerates atherosclerosis. This may have important implications for future therapeutic interventions targeting inflammatory pathways. The social and economic impacts of atherosclerosis and subsequent macrovascular clinical manifestations like myocardial infarction (MI) and stroke are immense. Complications of atherosclerosis and MI, such as heart failure, and also recurrent cardiovascular events present a major challenge. They seem to occur more frequently after MI than expected by presumed linear progression of atherosclerosis. In a recent article in Nature , Dutta et al1 found strong evidence that, like in a vicious cycle, acute MI itself accelerates atherosclerosis by triggering a burst of acute systemic inflammation initiated by progenitor cell mobilization from the bone marrow niche. The authors present 4 major findings: (1) acute MI (but also stroke) activates the sympathetic nervous system (SNS); (2) this causes the release of upstream progenitor cells from bone marrow niches; (3) these cells are hosted by the spleen leading to amplified extramedullary myelopoiesis; and (4) at the end of this multimodal cascade, there is a strong increase in inflammation in atherosclerotic plaques, leading to increased size and instability. The authors also show that experimental MI in apolipoprotein E–/– mice activates inflammatory pathways. In serial measurements after coronary ligation, concentrations of cathepsin in aortic plaques and the expression of the inflammatory cytokines interleukin (IL)-6, myeloperoxidase, and matrix metalloproteinase-9 were increased. Consecutively, this was associated with higher vulnerability of atherosclerotic plaque, as demonstrated by decreased thickness of the fibrous cap, and enlarged necrotic cores. Moreover, after MI, the number of macrophages and monocytes, especially the Ly-6Chigh monocyte subset, were increased in the aorta of apolipoprotein E–/– mice and exhibited higher levels of messenger …