Osteopontin Concentrations Research Articles

Plasma Concentrations of New Biochemical Markers of Atherosclerosis in Patients with Dyslipidemia-A Pilot Study.

Background and Objectives: The process of atherosclerotic plaque formation and its destabilisation is a process in which many proteins and cytokines are involved. Examples of such proteins are osteopontin (OPN), osteoprotegerin (OPG), metalloproteinases (MMPs) and myeloperoxidase (MPO). The aim of our study is to compare the concentrations of the above-mentioned markers in the plasma of patients with the confirmed presence of rupture plaque in comparison with the plasma of healthy people. Materials and Methods: The study included people suffering from dyslipidemia in whom the presence of unstable atherosclerotic plaque was confirmed by ultrasound. The concentrations of OPN, OPG, MPO, metalloproteinase 2 (MMP-2), and metalloproteinase 9 (MMP-9) in the plasma of these people were determined and compared with the concentrations of these proteins in the plasma of healthy people. Results: Levels of MMP-2, MMP-9 (p < 0.001), OPN, and OPG (p < 0.05) were statistically significantly lower in the group of healthy people than in the study group. Differences in MPO concentration were not statistically significant (p = 0.073). Conclusions: In the plasma of people with confirmed presence of rupture plaque, the concentrations of OPN, OPG, and MMPs are higher compared to the group of healthy people, which may suggest the use of these proteins as novel markers of the presence of unstable atherosclerotic plaque.

A Decreased Absolute Number of Treg Cells in Patients with Active Rheumatoid Arthritis is Associated with Elevated Serum Osteopontin Levels with Disease Progression.

Rheumatoid arthritis (RA) is a chronic and refractory autoimmune disease characterized by synovial inflammation with unknown aetiology. Immune system dysfunction mediated by CD4+ Tlymphocytes, which is regulated by the cytokine osteopontin (OPN), plays an important role in the pathogenesis of RA. In this study, the levels of peripheral CD4+ Tsubsets and serum OPN in patients with active RA were measured and analysed to determine the possible pathogenesis of RA and to provide potential therapeutic targets. Serum OPN levels in both patients with active RA and patients with refractory RA were higher than those in healthy controls (HCs). Compared with HCs, the absolute numbers of Th2 cells increased in patients with active RA, while the absolute counts of Th1 and Treg cells decreased. There was no significant difference in CD4+ Tsubset levels between new-onset and refractory patients. As the condition persisted or deteriorated, a gradual increase in the levels of OPN and gradual declines in the absolute counts of Th1 and Treg cells were observed in patients with active RA. The fewest Th1 and Treg cells and the highest OPN levels were observed in patients with high disease activity. The serum OPN level was only significantly negatively correlated with the absolute counts of Treg cells in the CD4+ Tlymphocyte subsets. Fewer Treg cells with the increase in disease activity may be related to the increased OPN concentration, which may provide new ideas and directions for the targeted immunoregulatory treatment of RA.

Osteopontin-induced vascular hyperpermeability through tight junction disruption in diabetic retina

Diabetic retinopathy is a major cause of blindness in developed countries, and is characterized by deterioration of barrier function causing vascular hyperpermeability and retinal edema. Vascular endothelial growth factor (VEGF) is a major mediator of diabetic macular edema. Although anti-VEGF drugs are the first-line treatment for diabetic macular edema, some cases are refractory to anti-VEGF therapy. Osteopontin (OPN) is a phosphoglycoprotein with diverse functions and expressed in various cells and tissues. Elevated OPN level has been implicated in diabetic retinopathy, but whether OPN is involved in hyperpermeability remains unclear. Using streptozotocin-induced diabetic mice (STZ mice) and human retinal endothelial cells (HRECs), we tested the hypothesis that up-regulated OPN causes tight junction disruption, leading to vascular hyperpermeability. The serum and retinal OPN concentrations were elevated in STZ mice compared to controls. Intravitreal injection of anti-OPN neutralizing antibody (anti-OPN Ab) suppressed vascular hyperpermeability and prevented decreases in claudin-5 and ZO-1 gene expression levels in the retina of STZ mice. Immunohistochemical staining of retinal vessels in STZ mice revealed claudin-5 immunoreactivity with punctate distribution and attenuated ZO-1 immunoreactivity, and these changes were prevented by anti-OPN Ab. Intravitreal injection of anti-OPN Ab did not change VEGF gene expression or protein concentration in retina of STZ mice. In an in vitro study, HRECs were exposed to normal glucose or high glucose with or without OPN for 48 h, and barrier function was evaluated by transendothelial electrical resistance and Evans blue permeation. Barrier function deteriorated under high glucose condition, and was further exacerbated by the addition of OPN. Immunofluorescence localization of claudin-5 and ZO-1 demonstrated punctate appearance with discontinuous junction in HRECs exposed to high glucose and OPN. There were no changes in VEGF and VEGF receptor-2 expression levels in HRECs by exposure to OPN. Our results suggest that OPN induces tight junction disruption and vascular hyperpermeability under diabetic conditions. Targeting OPN may be an effective approach to manage diabetic retinopathy.

Osteopontin-A Potential Biomarker for IgA Nephropathy: Machine Learning Application.

Many potential biomarkers in nephrology have been studied, but few are currently used in clinical practice. One is osteopontin (OPN). We compared urinary OPN concentrations in 80 participants: 67 patients with various biopsy-proven glomerulopathies (GNs)—immunoglobulin A nephropathy (IgAN, 29), membranous nephropathy (MN, 20) and lupus nephritis (LN, 18) and 13 with no GN. Follow-up included 48 participants. Machine learning was used to correlate OPN with other factors to classify patients by GN type. The resulting algorithm had an accuracy of 87% in differentiating IgAN from other GNs using urinary OPN levels only. A lesser effect for discriminating MN and LN was observed. However, the lower number of patients and the phenotypic heterogeneity of MN and LN might have affected those results. OPN was significantly higher in IgAN at baseline than in other GNs and therefore might be useful for identifying patients with IgAN. That observation did not apply to either patients with IgAN at follow-up or to patients with other GNs. OPN seems to be a valuable biomarker and should be validated in future studies. Machine learning is a powerful tool that, compared with traditional statistical methods, can be also applied to smaller datasets.

The value of plasma hypoxia markers for predicting imaging-based hypoxia in patients with head-and-neck cancers undergoing definitive chemoradiation

BackgroundTumor hypoxia worsens the prognosis of head-and-neck squamous cell carcinoma (HNSCC) patients, and plasma hypoxia markers may be used as biomarkers for radiotherapy personalization. We therefore investigated the role of the hypoxia-associated plasma proteins osteopontin, galectin-3, vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) as surrogate markers for imaging-based tumor hypoxia. MethodsSerial blood samples of HNSCC patients receiving chemoradiation within a prospective trial were analyzed for osteopontin, galectin-3, VEGF and CTGF concentrations. Tumor hypoxia was quantified in treatment weeks 0, 2 and 5 using [18F]FMISO PET/CT. The association between PET-defined hypoxia and the plasma markers was determined using Pearson’s correlation analyses. Receiver-operating characteristic analyses were conducted to reveal the diagnostic value of the hypoxia markers. ResultsBaseline osteopontin (r = 0.579, p < 0.01) and galectin-3 (r = 0.429, p < 0.05) correlated with the hypoxic subvolume (HSV) prior to radiotherapy, whereas VEGF (r = 0.196, p = 0.36) and CTGF (r = 0.314, p = 0.12) showed no association. Patients with an HSV > 1 mL in week 2 exhibited increased VEGF (p < 0.05) and CTGF (p < 0.05) levels in week 5. Pretherapeutic osteopontin levels were higher in patients exhibiting residual hypoxia at the end of treatment (104.7 vs. 60.8 ng/mL, p < 0.05) and could therefore predict residual hypoxia (AUC = 0.821, 95% CI 0.604–1.000, p < 0.05). ConclusionIn this exploratory analysis, osteopontin correlated with the initial HSV and with residual tumor hypoxia; therefore, there may be a rationale to study hypoxic modification based on osteopontin levels. However, as plasma hypoxia markers do not correspond to any spatial information of tumor hypoxia, they have limitations regarding the replacement of [18F]FMISO PET-based focal treatments. The results need to be validated in larger patient cohorts to draw definitive conclusions.

Qualitative and Quantitative Estimation of Osteopontin in Bull Semen and its Effect on Sperm Quality Parameters and Fertility Rate In vivo

Background: Osteopontin plays a crucial role in fertilization and subsequent embryonic development. Methods: The fresh semen samples were collected from crossbred Jersey and Haryana breeding bulls. The proteins in the seminal plasma and sperm membrane were precipitated and analysed by SDS-PAGE followed by western blotting. Based on the concentration of osteopontin in the seminal plasma, the bulls were grouped as high (≥20 pg/ml), medium (10-20 pg/ml) and low (≤10 pg/ml) OPN groups. Result: Total 13 and 14 different proteins were identified in the seminal plasma and sperm membrane, respectively. The sperm velocity parameters (straight line velocity and straightness) and functional membrane integrity showed significant difference among high, medium and low OPN groups. Significant difference (P less than 0.05) was observed in malondialdehyde, SOD and glutathione reductase levels. Bulls which are having high osteopontin concentration in the semen had less sperm abnormality, less apoptotic cells, better resistance to cryopreservation, less damaged chromatin and higher conception rate (64.80±1.09) than bulls having low osteopontin concentration in their semen. Therefore, osteopontin may be a valuable protein marker to predict fertility in bulls.

A Retrospective Cohort Study of the Association between Serum Osteopontin Levels and Aortic Stiffness in Hypertensive Patients.

By suppressing mineralization and preventing ectopic calcium deposits, osteopontin (OPN) has an inhibitory effect on vascular calcification. Also, there is an association between OPN and aortic stiffness (AS). We aimed to investigate the association between serum OPN levels and AS measured by carotid–femoral pulse wave velocity (cfPWV) in hypertensive patients. Baseline characteristics and fasting blood sampling of 120 participants with hypertension and 120 participants without hypertension were acquired. Serum OPN concentrations were determined by enzyme-linked immunosorbent assay. In total, 43 (35.9%) participants were assigned to the AS group with cfPWV of >10 m/s in hypertensive patients. There were more patients with diabetes mellitus, old age, high systolic blood pressure, high serum intact parathyroid hormone (iPTH), elevated C-reactive protein, and high OPN levels in the AS group compared with the control group in hypertensive participants. A multivariate logistic regression analysis discloses that age, SBP, serum OPN, and iPTH levels were independently associated with AS in hypertensive patients. Moreover, according to a multivariate forward stepwise linear regression analysis, OPN level is positively associated with cfPWV. In conclusion, serum OPN level is assumed to be a potential biomarker to predict AS and is positively associated with cfPWV in patients with hypertension.

Concentration of osteopontin in human milk and associated factors in Chinese populations from 2011 to 2013

To analyze content of human milk osteopontin(OPN) and to explore associated factors in Chinese populations. The samples and data were extracted from the database for human milk composition in China between 2011 and 2013. A sub-sample of 459 mothers was randomly selected after stratification according to lactation stage, and human milk OPN concentrations were determined by ultra performance liquid chromatography-tandem mass spectrometer(UPLC/MS). The average OPN concentration(M(P25, P75)) in breast milk was 44.0(30.1-72.0) mg/L within 0-330 days postpartum. OPN concentrations were independent of lactation stage, which were 45.6(31.8, 80.7) mg/L in colostrum, 41.3(29.2, 70.0) mg/L in transitional milk and 46.9(30.2, 71.9) mg/L in mature milk, corresponding to 0.40%、0.42% and 0.65% of the total milk protein content(OPN/protein%). The percentage of OPN to total protein in milk showed an increasing trend with lactation progression(r=0.21, P&lt;0.001). Multivariate analysis showed that sleep quality of mothers within one week prior to milk collection was correlated with the breast milk OPN level(P=0.04). The OPN level in breast milk from mothers with good sleep quality was significantly higher than that from mothers with poor sleep quality(46.5 mg/L vs.34.7 mg/L). The median level of milk OPN concentration in mothers from Yunnan was higher than mothers from Beijing(50.5 mg/L vs.36.1 mg/L, P=0.03). Maternal age, mode of delivery, prepregnancy body mass index, weight gain during pregnancy, passive smoking and outdoor activities 24 hours prior to milk collection were not correlated with milk OPN concentration. OPN concentration in breast milk was not related to preterm birth. Also, milk OPN concentration did not correlate with diarrhoea, respiratory disease, or allergic disease in infants during two weeks before milk collection. The concentration of OPN in breast milk of Chinese woman may be similar among different lactation stages. Geographic region and sleep quality of mothers may be related to the milk OPN concentration.

Determination of osteopontin and antioxidant concentrations from epididymal fluids in mongrel dogs

The extracellular matrix phosphoprotein, Osteopontin (OPN) has been found in tissue fluids and organs, including the male reproductive tract. The vesicular gland and the ampullary region in the male reproductive system have been documented to secrete OPN, a 55 kDa fertility associated protein. The objective of this study was to quantify OPN and antioxidant concentrations in different epididymal sections in mongrel dogs. Current results showed a significantly (p&lt;0.01) higher concentration of OPN in cauda epididymal fluid than in the caput. Glutathione peroxidase (GPx) and Catalalse (CAT) concentrations were also higher in the cauda epididymal segment than in the caput. To the author’s best knowledge, this is the first report on quantification of OPN levels in the epididymal fluid harvested from the different sections of epididymis. Further studies should focus on correlation and association of this protein from the different epididymal segments with seminal attributes.

Osteopontin levels are associated with late-time lower regional brain volumes in multiple sclerosis

Osteopontin (OPN) is a proinflammatory marker produced by systemic immune and central nervous system (CNS) resident cells. We examined, if the level of OPN in the cerebrospinal fluid (CSF) and blood is associated with late-time regional brain volumes and white matter (WM) lesion load in MS. Concentrations of OPN in blood and CSF were related to MRI findings 10.1 ± 2.0 years later in 46 patients with MS. OPN concentration was measured by ELISA, while regional brain volumes and lesion load was assessed by magnetic resonance imaging (MRI) using 3D MPRAGE sequence and automated MR volumetry. OPN measured in the CSF was associated with several regional brain volumes and WM lesion load measured 10.1 ± 2.0 years later. CSF OPN concentration correlated with long-term enlargement of lateral- and inferior lateral ventricles and the elevation of gross CSF volume, in conjunction with the reduction of several cortical/subcortical gray matter and WM volumes. Serum OPN showed no long-term association with regional brain volumes. OPN measured from the CSF but not from the serum was associated with lower regional brain volumes measured a decade later, indicating the primary role of inflammation within the CNS in developing long-term brain related alterations.

Efficacy of nanomicelle curcumin, Nigella sativa oil, and their combination on bone turnover markers and their safety in postmenopausal women with primary osteoporosis and osteopenia: A triple-blind randomized controlled trial.

Literature supports the potential effects of nanomicelle curcumin and Nigella sativa on the amelioration of osteoporosis, a health concern of postmenopausal women. This study aimed to evaluate the impacts of nanomicelle curcumin (CUR), Nigella sativa oil (NS), and their combination on bone turnover biomarkers and assess their safety. This triple‐blind randomized controlled trial was performed on 120 postmenopausal women aged 50–65 with primary osteoporosis or osteopenia. The subjects were randomly allocated to receive microcrystalline cellulose (placebo), 80 mg of CUR, 1000 mg of NS, or their combination (CUR‐NS) for 6 months. All patients were also treated with alendronate (70 mg) and calcium (500 mg), vitamin D (400 IU) supplements. The serum levels of alkaline phosphatase (ALP), osteocalcin (OC), and osteopontin (OP) were measured at the baseline and after the intervention. For safety assessment, the hepatic enzyme levels of aspartate transaminase and alanine transaminase as well as serum urea and creatinine were evaluated. ALP levels were significantly reduced in the NS (p = .029) and CUR‐NS (p = .015) groups compared with those in the placebo. After adjustment for the covariates, this effect was still significant in the CUR‐NS group (p = .004). The OC levels were decreased in the placebo, CUR, and NS groups, and the OP concentration also was attenuated in all groups through the trial. However, the intergroup differences were not significant for both biomarkers. Evaluating the key renal metabolites and hepatic enzyme levels indicated no toxicity of the administered doses. This study reveals the beneficial effects of CUR‐NS on the improvement of some bone turnover biomarkers. These compounds seem to be safe at the current dosage for supplementation in postmenopausal women.

Nucleation kinetics of calcium oxalate monohydrate as a function of pH, magnesium, and osteopontin concentration quantified with droplet microfluidics.

A droplet-based microfluidic platform is presented to study the nucleation kinetics of calcium oxalate monohydrate (COM), the most common constituent of kidney stones, while carefully monitoring the pseudo-polymorphic transitions. The precipitation kinetics of COM is studied as a function of supersaturation and pH as well as in the presence of inhibitors of stone formation, magnesium ions (Mg2+), and osteopontin (OPN). We rationalize the trends observed in the measured nucleation rates leveraging a solution chemistry model validated using isothermal solubility measurements. In equimolar calcium and oxalate ion concentrations with different buffer solutions, dramatically slower kinetics is observed at pH 6.0 compared to pHs 3.6 and 8.6. The addition of both Mg2+ and OPN to the solution slows down kinetics appreciably. Interestingly, complete nucleation inhibition is observed at significantly lower OPN, namely, 3.2 × 10-8 M, than Mg2+ concentrations, 0.875 × 10-4 M. The observed inhibition effect of OPN emphasizes the often-overlooked role of macromolecules on COM nucleation due to their low concentration presence in urine. Moreover, analysis of growth rates calculated from observed lag times suggests that inhibition in the presence of Mg2+ cannot be explained solely on altered supersaturation. The presented study highlights the potential of microfluidics in overcoming a major challenge in nephrolithiasis research, the overwhelming physiochemical complexity of urine.

Osteopontin and malaria: no direct effect on parasite growth, but correlation with P. falciparum-specific B cells and BAFF in a malaria endemic area

BackgroundThe dysregulation of B cell activation is prevalent during naturally acquired immunity against malaria. Osteopontin (OPN), a protein produced by various cells including B cells, is a phosphorylated glycoprotein that participates in immune regulation and has been suggested to be involved in the immune response against malaria. Here we studied the longitudinal concentrations of OPN in infants and their mothers living in Uganda, and how OPN concentrations correlated with B cell subsets specific for P. falciparum and B cell activating factor (BAFF). We also investigated the direct effect of OPN on P. falciparum in vitro.ResultsThe OPN concentration was higher in the infants compared to the mothers, and OPN concentration in infants decreased from birth until 9 months. OPN concentration in infants during 9 months were independent of OPN concentrations in corresponding mothers. OPN concentrations in infants were inversely correlated with total atypical memory B cells (MBCs) as well as P. falciparum-specific atypical MBCs. There was a positive correlation between OPN and BAFF concentrations in both mothers and infants. When OPN was added to P. falciparum cultured in vitro, parasitemia was unaffected regardless of OPN concentration.ConclusionsThe concentrations of OPN in infants were higher and independent of the OPN concentrations in corresponding mothers. In vitro, OPN does not have a direct effect on P. falciparum growth. Our correlation analysis results suggest that OPN could have a role in the B cell immune response and acquisition of natural immunity against malaria.

Остеопонтин та остеокальцин при раку легенів

Актуальність. Рак легенів (РЛ) займає лідуючу позицію в структурі онкологічної захворюваності та є однією з провідних причин смерті від онкозахворювань. Пошук ефективних підходів до оцінки перебігу РЛ з використанням інформативних критеріїв є актуальним завданням сучасної онкології. РЛ зазвичай перебігає з підвищеним рівнем у крові остеопонтину (ОР) та остеокальцину (ОС), що має прогностичну значущість у рамках виживання хворих. Сироватковий вміст цих остеоонкомаркерів прямо асоціюється з наявністю у хворих метастазів і з високими темпами прогресування захворювання, але ці дані вимагають подальшого уточнення. Мета дослідження: оцінити клініко-патогенетичну й прогностичну значущість ОР та ОС у хворих із різними варіантами перебігу РЛ. Матеріали та методи. Обстежено 115 хворих на РЛ віком від 24 до 80 років (у середньому 58 років), серед яких було 78 % чоловіків і 22 % жінок. Жоден пацієнт раніше не був прооперований із приводу РЛ і до обстеження не отримував радіохіміотерапію. Дрібноклітинний гістологічний варіант РЛ встановлений у 17 % від числа хворих, недрібноклітинний — у 83 %. Імуноферментним аналізом вивчали в сироватці крові показники ОР, ОС і судинного ендотеліального фактору росту (VEGF). Результати. У хворих на РЛ збільшуються в сироватці крові показники онкоостеоасоційованих маркерів OP та OC, що спостерігається відповідно в 99 і 98 % випадків. Зміни ОР і ОС пов’язані з формою захворювання, гістологічним його варіантом, ступенем диференціації, інтегральною тяжкістю перебігу РЛ, характером ускладнень первинної пухлини й особливостями метастазування. Високий рівень маркерів у крові мало впливає на трирічне виживання пацієнтів, а прогнознегативними критеріями відносно перебігу РЛ є параметри ОР. Показники ОР і ОС у крові між собою прямо корелюють, а вміст ОР, окрім того, має позитивні співвідношення з таким пухлинним маркером, як VEGF, що належить до стимуляторів неоангіогенезу. Метастатичне ураження скелету й розвиток порушень мозкового кровообігу внаслідок подальшої радіохіміотерапії прямо пов’язані з високою концентрацією в крові ОР, який також є чинником ризику інших ускладнень лікування. Висновки. Підвищені рівні в крові ОР і ОС мають важливе клініко-патогенетичне значення при РЛ, а вивчення показників цих остеоонкомаркерів сприятиме ранній діагностиці окремих ознак перебігу захворювання, підвищенню якості оцінки метастазування в кістково-суглобовий апарат і прогнозуванню можливих ускладнень радіохіміотерапії.

The role of calcium metabolism disorders in the formation of different density calculi with calcium oxalate nephrolithiasis

Objective To investigate the relationship between calcium metabolism disorders, stone formation inhibitor levels and stone density in primary and recurrent calcium-oxalate nephrolithiasis.Material and Methods Sixty nine patients with urolithiasis were examined, their average age was 41,4 ± 9,5 years. Two main groups were distinguished: Group 1 – primary calcium-oxalate nephrolithiasis (PN), Group 2 – recurrent calcium-oxalate nephrolithiasis (RN). Then each group was divided into two subgroups – A and B according to stone density: 500–1000 HU and from 1000–1500 HU, respectively. Stone density was determined by computed tomography (CT). PTH (parathormone), PTHrP (parathyroid hormone related protein), vitamin D, total blood calcium (Ca), ionized blood Ca, total blood protein, Ca and urine pH were also examined. After the examination, patients underwent surgical removal of the stones.Results It was found that 41.9% of group 1 and 46.9% of group 2 patients had grade I obesity. Average creatinine level in group 2 was 9.7% higher than in group 1 (p &lt; 0.05). Urea level in both groups was not statistically significantly different. Glomerular filtration rate (GFR) was comparable. Groups 2A and 2B had higher PTHrP values (77.61 and 76.98 pg/mL, respectively) combined with relatively high PTH levels (2A – 4.4 pg/mL and 2B – 5.1 pg/mL), relatively low osteopontin concentration (2A – 0.044 pg/ mL, 2B – 1.106 pg/mL), compared to those in group 1 (p &lt; 0.05). Pairwise unidirectional differences between groups 1A and 2A, 1B and 2B were found to correlate positively with density values: for osteopontin: r = 0.992 (p &lt; 0.05); for vitamin D: r = 0.831 (p &lt; 0.05); for blood Ca2+ ions: r = 0.836 (p &lt; 0.05); for urine pH: r = 0.863 (p &lt; 0.05). There was a negative correlation with the daily concentration of urinary calcium ions with the density of concrements: r = -0.663; p &lt; 0.05. The concentration of osteopontin was significantly higher in Group 1B and 2B patients, and it was significantly lower in patients with stones of &lt; 1000 HU density. Higher values of osteopontin concentration were noted in groups 1B and 2B in relation to groups 1A (p &lt; 0.05) and 2A (p &lt; 0.05). The increase of blood Ca2+ ions in patients in groups 1B and 2B in relation to groups 1A (p &lt; 0.05) and 2A (p &lt; 0.05) was also accompanied by higher values of vitamin D.Conclusion Patients with denser stones showed high values of osteopontin and PTHrP in serum and low values of urinary calcium ions, which may lead to the formation of concrements on the matrix with an organic base. Determination of calcium metabolism makes it possible to predict recurrence of KSD in primary calcium oxalate nephrolithiasis and assess the severity of mineral metabolism disorders in recurrent calcium oxalate nephrolithiasis.

Matrix and Sampling Effects on Quantification of Protein Biomarkers of Drug-Induced Liver Injury.

Macrophage colony stimulating factor 1 receptor (MCSF1R), osteopontin (OPN), high-mobility group protein B1 (HMGB1), glutamate dehydrogenase (GLDH), keratin 18 (K18), and caspase-cleaved keratin 18 (ccK18) are considered promising mechanistic biomarkers for the diagnosis of drug-induced liver injury. Here, we aim to elucidate the impact of the sample matrix and handling on the quantification of these emerging protein biomarkers. We investigated effects such as time from collection to centrifugation during serum (± gel) or EDTA plasma preparation on two assay platforms: immunoaffinity liquid chromatography mass spectrometric assays and sandwich immunoassays. Furthermore, we measured GLDH activity with an enzymatic activity assay. Matrix effects were observed particularly for HMGB1 and MCSF1R. HMGB1 levels were higher in serum than in plasma, whereas higher concentrations of MCSF1R were observed in plasma than in serum. A comparison of sample collection to centrifugation time ranging from 15 to 60 min demonstrated increasing levels of HMGB1 in serum, while MCSF1R, OPN, GLDH, and ccK18 concentrations remained stable. Additionally, there was a poor correlation in HMGB1 and ccK18 levels between serum and plasma. Considering the observed matrix effects, we recommend plasma as a matrix of choice and cross-study comparison studies to be limited to those using the same matrix.

Кальцитонин, остеокальцин и остеопонтин при раке легкого

Актуальность. Маркерами рака легкого являются низкие уровни в крови гормона кальцитонина, а высокие показатели белков остеокальцина и остеопонтина, связанные между собой, отражают процессы костного метастазирования и обладают прогностической значимостью. Цель исследования: оценить уровни в крови кальцитонина, остеокальцина и остеопонтина, факторы, их определяющие, характер взаимосвязей и прогностическую роль в костном метастазировании при разных вариантах течения рака легкого. Материалы и методы. Обследованы 86 больных раком легкого в возрасте от 24 до 80 лет (в среднем 58 лет), среди которых было 84 % мужчин и 16 % женщин. Центральная форма рака имела место в 81 % наблюдений, периферическая — в 19 %. Мелкоклеточный гистологический вариант заболевания установлен у 21 % от числа больных, немелкоклеточный — у 79 %, в том числе аденокарцинома — у 38 %, плоскоклеточная карцинома — у 35 %, крупноклеточная карцинома — у 6 %. IIIA стадия опухолевого процесса отмечена в 12 % случаев, IIIB — в 21 %, IV — в 67 %. Результаты. Рак легкого сопровождается уменьшением в крови у 20 % больных остеоассоциированного гормона кальцитонина на фоне повышения концентраций опухолевых маркеров остеокальцина и остеопонтина (соответственно в 76 и 79 % наблюдений), что тесно связано с морфологическим вариантом (мелко- и немелкоклеточный), стадийностью и тяжестью течения патологического процесса, определяет развитие компрессионного синдрома, прорастание опухоли в грудную стенку, метастазирование в лимфатические узлы и кости, возникновение осложнений химиорадиотерапии в виде лучевого пневмофиброза, легочной гипертензии и миелодепрессии, а показатели остеокальцин- и остеопонтинемии обратно коррелируют с продолжительностью жизни больных. Выводы: кальцитонин, остеокальцин и остеопонтин при раке легкого являются маркерами тяжести течения заболевания, отражают метастазирование опухолевого процесса в скелет и обладают прогностической значимостью.

Osteopontin Levels in Human Milk Are Related to Maternal Nutrition and Infant Health and Growth

Background: Osteopontin (OPN) is a glycosylated phosphoprotein found in human tissues and body fluids. OPN in breast milk is thought to play a major role in growth and immune system development in early infancy. Here, we investigated maternal factors that may affect concentrations of OPN in breast milk, and the possible associated consequences for the health of neonates. Methods: General characteristics, health status, dietary patterns, and anthropometric measurements of 85 mothers and their babies were recorded antenatally and during postnatal follow-up. Results: The mean concentration of OPN in breast milk was 137.1 ± 56.8 mg/L. Maternal factors including smoking, BMI, birth route, pregnancy weight gain, and energy intake during lactation were associated with OPN levels (p < 0.05). Significant correlations were determined between body weight, length, and head circumference, respectively, and OPN levels after one (r = 0.442, p = < 0.001; r = −0.284, p = < 0.001; r = −0.392, p = < 0.001) and three months (r = 0.501, p = < 0.001; r = −0.450, p = < 0.001; r = −0.498, p = < 0.001) of lactation. A negative relation between fever-related infant hospitalizations from 0–3 months and breast milk OPN levels (r = −0.599, p < 0.001) was identified. Conclusions: OPN concentrations in breast milk differ depending on maternal factors, and these differences can affect the growth and immune system functions of infants. OPN supplementation in infant formula feed may have benefits and should be further investigated.

Osteopontin and galectin-3 as biomarkers of maladaptive right ventricular remodelingin pulmonary hypertension.

Aim: This study assessed the utility of osteopontin (OPN) and galectin-3 (Gal-3) as biomarkers of maladaptive right ventricular remodeling in pulmonary hypertension (PH). Materials & methods: We examined plasma levels of OPN and Gal-3 in patients with PH (n=62), dilated cardiomyopathy (n=34), left ventricular hypertrophy (LVH;n=47), and controls without right ventricle (RV) or LV abnormalities (n=38). Results: OPN and Gal-3 levels were higher in PH, dilated cardiomyopathyand LVH than in the controls. OPN concentrations in PH patients with maladaptive RV were significantly higher than in those with adaptive RV. Gal-3 did not differentiate between adaptive and maladaptive RV remodeling in PH. OPN and Gal-3 levels did not correlate with parameters of LV remodeling. Conclusion: OPN is a potential biomarker of RV maladaptation.

Evaluation of local and circulating osteopontin in malignant and benign primary bone tumors

PurposeThe development of novel and efficient biomarkers for primary bone cancers is of grave importance. MethodsThe expression pattern of osteopontin (OPN) was investigated in the 153 patients with benign (n = 72) and malignant (n = 81) primary bone cancers. Both local and circulating OPN mRNA expression levels and their protein concentration in serum and tumor site were assessed using real-time qRT-PCR, ELISA, and immunohistochemistry techniques, respectively. As a control, 29 healthy individuals were considered. The number of 153 tumor tissue specimens and the 153 paired margins were taken on surgical resection from the patients. 153 blood samples were also drained from all participants, then peripheral blood mononuclear cells (PBMC) and sera were separated. ResultsThe mean mRNA expression was significantly higher in all of the cancerous tissues than the paired margins and the PBMC of the patients than the controls. Consistently, the protein concentrations of OPN in serum and tumor tissues were significantly higher in the patients. Furthermore, the malignant cases had significantly elevated the mRNA levels and the protein compared to the benign cases. OPN could potentially differentiate the patients from the controls with 100% sensitivity and specificity in serum. Moreover, OPN could predict some of the malignant cases' clinicopathological features, including metastasis, recurrence, grade, and response to chemotherapy. ConclusionsIn conclusion, OPN might be involved in the pathogenesis of primary bone tumors and can be considered as a potential biomarker to bone cancer diagnosis.