Aluminum (Al) is recognized worldwide as serious inorganic contaminants. Exposure to Al is associated with low BMD and an increased risk of osteoporosis. However, the precise molecular mechanisms remains unclear. Thus, in this study, rats were orally exposed to 0 (control group, CG) and 0.4g/L AlCl3 (AlCl3 treated group, AG) in drinking water for 120days; osteoblasts were treated with AlCl3 (0.12mg/mL) and/or TGF-β1 (4.5ng/mL) for 24h. We found that AlCl3 decreased the BMD, damaged femoral ultrastructure, decreased the activities of GSH-Px and SOD, and increased the levels of ROS and MDA in bone, decreased the activity of B-ALP and content of PINP, and increased the activity of TRACP-5b and content of NTX-I in serum, decreased mRNA expressions of TGF-β1, TβRI, TβRII and Smad4, protein expressions of TGF-β1, p-Smad2/3 and Smad2/3/4 complex, and increased Smad7 mRNA expression in bone and in osteoblasts. Moreover, we found exogenous TGF-β1 application reversed the inhibitory effect of AlCl3 on osteoblasts activity by activating the TGF-β1/Smad signaling pathway and increasing the mRNA expressions of ALP and Col I in osteoblasts. These results demonstrate that AlCl3 induces bone impairment through inactivation of TGF-β1/Smad signaling pathway.

The ovariectomized old cynomolgus monkey is a recognized model of human osteoporosis, and the same species can be used for the assessment of the efficacy and potential toxicity of agents intended to prevent or treat osteoporosis. Several assays have been developed that can measure the same biochemical markers of bone turnover as are used in human patients for the diagnosis and treatment follow-up of bone-related diseases, including osteoporosis. The aim of the present study was to describe the results obtained with these assays in normal control monkeys, their variations with age and sex, and their sensitivity in monitoring the bone turnover induced by ovariectomy in old skeletally mature cynomolgus monkeys. Seven old cynomolgus monkeys were bilaterally ovariectomized and 13 age-matched monkeys were sham-operated. Bone mineral density and biochemical markers were measured before and at regular intervals after surgery for up to 20 months. Total alkaline phosphatase (total ALP), bone-specific alkaline phosphatase isoenzyme (bone ALP) and osteocalcin (OC) were highly correlated to the decrease in bone mineral density (BMD) induced by ovariectomy. Deoxypyridinoline (DPD) measured by enzyme-linked immunoassay was insensitive to the bone resorption induced by ovariectomy, but cross-linked N-telopeptide (NTX-I) was higher in ovariectomized monkeys than in control monkeys. These results demonstrate that reliable biochemical parameters are available to adequately monitor and provide insight into osteoclastic bone resorption and osteoblastic bone formation, the two components of bone turnover in this animal model, and can thus be used to assess the efficacy and toxicity of potential therapeutic agents.

OBJECTIVE

Associations between dietary factors and bone metabolism in Asians have not been thoroughly investigated. The purpose of this study was to determine the effect of dietary intake of calcium (Ca), phosphorus (P), sodium (Na), and protein on bone metabolism in elderly Japanese women by the duplicate portion sampling method.

METHODS

The subjects were 43 healthy female volunteers (mean age, 68.3 y; standard deviation, 6.8). Dietary nutrients were directly determined by using a 24-h duplicate meal portion. Serum osteocalcin and bone alkaline phosphatase and urinary deoxypyridinoline (DPD) and type I collagen cross-linked N-telopeptides (NTX-I) were measured as markers of bone turnover. Hormones related to bone metabolism, including serum 25-hydroxyvitamin D(3), 1,25-dihydroxyvitamin D(3), and intact parathyroid hormone also were determined. Urinary parameters were corrected for urinary creatinine concentration.

RESULTS

The mean (standard deviation) daily dietary intakes of Ca, P, Na, and protein were 660 (195) mg, 996 (208) mg, 4080 (1142) mg, and 63.9 (15.5) g, respectively. Dietary Ca was negatively correlated with urinary DPD (r = -0.417, P = 0.005) and NTX-I (r = -0.324, P = 0.034), and dietary P was negatively correlated with urinary DPD (r = -0.307, P = 0.045). Multiple regression analysis showed that only dietary Ca was associated with urinary DPD and NTX-I. Dietary Ca intake was significantly associated with bone resorption markers, but no other dietary factors were associated with any of the biochemical markers.

CONCLUSIONS

The major factor having an adverse effect on the bone health of the elderly Japanese women is low Ca intake and not other dietary minerals or protein.

Crystals of the neurotoxin-I (NTX-I) from the venom of the middle Asian cobra Naja naja oxiana have been grown by vapour diffusion and dialysis methods. The crystals belong to space group P2(1)2(1)2 with dimension of a = 25.19 A, b = 75.59 A, c = 36.09 A and diffract to 1.9 A resolution. The asymmetric unit contains one molecule (Vm = 2.2 A/Da). Using the molecule of alpha-cobratoxin (CTX) as a starting model for NTX-I structure determination coordinates of C alpha atoms of the NTX-I molecule were obtained and the position of NTX-I in the unit cell was derived.

Haemophilia A and B have been associated with increased prevalence of low bone mineral density (BMD). However, the utility of bone turnover markers (BTM) remains unknown. The aim of this study was to evaluate bone metabolism in men with haemophilia and to investigate associations between BTM and bone disease. Serum N- (NTX-I), C-terminal telopeptide of type I collagen (CTX-I) and tartrate-resistant acid phosphatase band-5b (TRAP-5b), as bone resorption markers, and osteocalcin (OC) and bone-specific alkaline phosphatase (b-ALP), as bone formation markers, were assessed. Seventy men with haemophilia A (n = 59) or B (n = 11) were studied. Patients with low BMD had significantly higher b-ALP concentrations compared with those with normal BMD (12.8 ± 1.60 vs. 9.72 ± 0.58 μg/L, P = 0.009), without any differences in the other BTM. NTX-I and CTX-I concentrations were negatively associated with oestradiol levels and hip BMD and positively with human immunodeficiency virus infection, number of affected joints and arthropathy scores. B-ALP and OC concentrations were negatively associated with hip BMD, severity of haemophilia and fracture history, and positively with the number of affected joints and testosterone concentrations. After multivariate analysis, NTX-I levels remained negatively associated with oestradiol levels, whereas b-ALP concentrations negatively correlated with the level of physical activity and positively with the number of affected joints. Increased bone metabolism exists in men with haemophilia and low BMD. Increased b-ALP levels may identify patients at high risk for fracture. Increased number of target joints, low physical activity and low oestradiol concentrations are independently associated with increased bone metabolism.

Purpose: Lipocalin 2 (LCN2) is an adipokine involved in many physiological functions, including bone metabolism. We previously demonstrated its implication in mouse models of mechanical unloading-induced osteoporosis and in a cohort of bed rest volunteers. We therefore aimed at studying its involvement in postmenopausal osteoporosis.

Methods: We measured serum LCN2 and correlated its levels to Dickkopf WNT Signaling Pathway Inhibitor 1 (DKK1), Tartrate Resistant Acid Phosphatase 5B (TRAcP5B), sclerostin, urinary N-terminal telopeptide of type I collagen (NTX), serum C-terminal telopeptide of type I collagen (CTX), parathyroid hormone and vitamin K by ELISA performed in a cohort of younger (50-65 years) and older (66-90 years) osteoporotic women in comparison to healthy subjects. A cohort of male healthy and osteoarthritic patients was also included. Sobel mediation analysis was used to test indirect associations among age, LCN2 and DKK1 or NTX.

<strong class="sub-t<em>i</em>tle"> Results: </strong> LCN2 levels were unchanged <em>i</em>n osteoporot<em>i</em>c and <em>i</em>n osteoarthr<em>i</em>t<em>i</em>s pat<em>i</em>ents when compared to healthy subjects and d<em>i</em>d not correlate w<em>i</em>th BMD. However, serum LCN2 correlated w<em>i</em>th age <em>i</em>n healthy women ((<em>i</em>)R</<em>i</em>) = 0.44; (<em>i</em>)P</<em>i</em>) = 0.003) and men ((<em>i</em>)R</<em>i</em>) = 0.5; (<em>i</em>)P</<em>i</em>) = 0.001) and serum concentrat<em>i</em>ons of DKK1 ((<em>i</em>)R</<em>i</em>) = 0.47; P = 0.003) and ur<em>i</em>nary <em>NTX</em> ((<em>i</em>)R</<em>i</em>) = 0.34; (<em>i</em>)P</<em>i</em>) = 0.04). Sobel med<em>i</em>at<em>i</em>on analys<em>i</em>s showed that LCN2 med<em>i</em>ates an <em>i</em>nd<em>i</em>rect relat<em>i</em>onsh<em>i</em>p between age and DKK1 ((<em>i</em>)P</<em>i</em>) = 0.02), but not w<em>i</em>th <em>NTX</em>, <em>i</em>n healthy subjects.

Conclusions: Taken together, the results suggest a hitherto unknown association between LCN2, DKK1 and age in healthy individuals, but not in postmenopausal osteoporotic women.

Keywords: BALP, bone-specific alkaline phosphatase; BMD, bone mineral density; BMI, body mass index; CTX, C-terminal telopeptide of type I collagen; DKK1; DKK1, Dickkopf WNT Signaling Pathway Inhibitor 1; IL, interleukin; LCN2, lipocalin 2; Lipocalin-2; NGAL; NTX, N-terminal telopeptide of type I collagen; NfκB, nuclear factor kappa-B; Osteoarthritis; Osteoporosis; PTH, parathyroid hormone; RANKL, receptor activator of nuclear factor kappa-B; TNF, tumor necrosis factor; TRAcP5B, tartrate-resistant acid phosphatase 5B; Wnt.

Objective: To determine the optimal combination of imaging and biochemical biomarkers to predict knee osteoarthritis (OA) progression.

Methods: Nested case-control study from the FNIH OA Biomarkers Consortium of participants with Kellgren-Lawrence grade 1-3 and complete biomarker data (n=539 to 550). Cases were knees with radiographic and pain progression between 24-48 months from baseline. Radiographic progression only was assessed in secondary analyses. Biomarkers (baseline and 24-month changes) with p<0.10 in univariate analysis were selected, including MRI (quantitative (Q) cartilage thickness and volume; semi-quantitative (SQ) MRI markers; bone shape and area; Q meniscal volume), radiographic (trabecular bone texture (TBT)), and serum and/or urine biochemical markers. Multivariable logistic regression models were built using three different step-wise selection methods (complex vs. parsimonious models).

Results: Among baseline biomarkers, the number of locations affected by osteophytes (SQ), Q central medial femoral and central lateral femoral cartilage thickness, patellar bone shape, and SQ Hoffa-synovitis predicted progression in most models (C-statistics 0.641-0.671). 24-month changes in SQ MRI markers (effusion-synovitis, meniscal morphology, and cartilage damage), Q central medial femoral cartilage thickness, Q medial tibial cartilage volume, Q lateral patellofemoral bone area, horizontal TBT (intercept term), and urine NTX-I predicted progression in most models (C-statistics 0.680-0.724). A different combination of imaging and biochemical biomarkers (baseline and 24-month change) predicted radiographic progression only, with higher C-statistics (0.716-0.832).

Conclusion: This study highlights the combination of biomarkers with potential prognostic utility in OA disease-modifying trials. Properly qualified, these biomarkers could be used to enrich future trials with participants likely to progress.

Keywords: MRI; biochemical markers; knee osteoarthritis; predictive validity.

There is a desperate need for the reliable detection of osteoarthritis (OA) at the early stage when patients are likely to benefit most from disease interventions. A variety of biochemical markers have been proposed, but their reliability varies among studies.

In this review, we aimed to answer the following questions: (1) are there biochemical markers that are differentially expressed in early OA versus healthy subjects, and (2) if so, what is the diagnostic value of these biomarkers for early OA?

Embase, PubMed, and Web of Science were searched to obtain all relevant studies up to March 2018, and studies comparing the biochemical markers between early OA and healthy controls were selected. The Downs and Black checklist was used to assess the risk of bias. Biomarkers that were investigated in five or more different populations were pooled for meta-analysis. A meta-regression analysis was performed to explore possible explanations for the heterogeneity of studies.

<em>I</em>n total, 26 articles met the criteria for the qualitative synthesis and 17 articles for the final quantitative synthesis. N-terminal crosslinked telopeptide of type <em>I</em> collagen (<em>NTX</em>-<em>I</em>) was the only biomarker found to be differently expressed in patients with early OA versus controls, without significant heterogeneity among studies (<em>I</em>2 = 0%, [Formula: see text] = 1.695, p = 0.792). The meta-regression analysis identified that sample size and affected joint possibly explained the heterogeneity among studies.

Although a wide range of biomarkers has been previously investigated in early OA, the diagnostic value of these biomarkers could not be determined because due to a low number of studies regarding any given biomarker. Large prospective and adequately powered studies are therefore required to validate these (and other) biomarkers for identifying early OA.

Ox<em>i</em>dat<em>i</em>ve stress <em>i</em>nduces damages of var<em>i</em>ous cell types or t<em>i</em>ssues through a repet<em>i</em>t<em>i</em>ve <em>i</em>mbalance between the system<em>i</em>c man<em>i</em>festat<em>i</em>on of react<em>i</em>ve oxygen spec<em>i</em>es (ROS) and detox<em>i</em>f<em>i</em>cat<em>i</em>on of the react<em>i</em>ve <em>i</em>ntermed<em>i</em>ates. Th<em>i</em>oacetam<em>i</em>de (TAA) <em>i</em>s well known for caus<em>i</em>ng several degenerat<em>i</em>ve d<em>i</em>seases by ox<em>i</em>dat<em>i</em>ve stress. However, study of the ant<em>i</em>ox<em>i</em>dant mechan<em>i</em>sms of stem cells <em>i</em>n TAA-<em>i</em>njured rat model <em>i</em>s <em>i</em>nsuff<em>i</em>c<em>i</em>ent. Therefore, we <em>i</em>nvest<em>i</em>gated the effect of placenta-der<em>i</em>ved mesenchymal stem cells (PD-MSCs) transplantat<em>i</em>on on l<em>i</em>ver and ovary of TAA-<em>i</em>njured rat models to study the ant<em>i</em>ox<em>i</em>dant effect <em>i</em>n degenerat<em>i</em>ve d<em>i</em>seases. In TAA-<em>i</em>njured rat model, PD-MSCs engrafted <em>i</em>nto damaged organ <em>i</em>nclud<em>i</em>ng l<em>i</em>ver and ovary <em>i</em>n PD-MSCs transplanted groups (Tx) compared w<em>i</em>th non-transplanted groups (<em>NTx</em>) (*(<em>i</em>)p</<em>i</em>)<0.05). Transplanted PD-MSCs reduced <em>i</em>nflammatory factors and upregulated ox<em>i</em>dat<em>i</em>ve stress factors <em>i</em>n Tx compared w<em>i</em>th <em>NTx</em> (*(<em>i</em>)p</<em>i</em>)<0.05). Also, transplanted PD-MSCs enhanced ant<em>i</em>ox<em>i</em>dants factors and organ funct<em>i</em>onal restorat<em>i</em>on factors <em>i</em>n Tx compared w<em>i</em>th <em>NTx</em>. These data show that PD-MSC transplantat<em>i</em>on tr<em>i</em>ggers the regenerat<em>i</em>on of organ (e.g., l<em>i</em>ver and ovary) damaged by ox<em>i</em>dat<em>i</em>ve stress from TAA treatment v<em>i</em>a act<em>i</em>vat<em>i</em>ng ant<em>i</em>ox<em>i</em>dant factors. Therefore, these data suggest the therapeut<em>i</em>c potent<em>i</em>al v<em>i</em>a ant<em>i</em>ox<em>i</em>dant effect and help understand the therapeut<em>i</em>c mechan<em>i</em>sm of PD-MSCs <em>i</em>n damaged t<em>i</em>ssues such as <em>i</em>n l<em>i</em>ver and reproduct<em>i</em>ve system.

Keywords: antioxidants; liver; ovary; placenta-derived mesenchymal stem cells; thioacetamide.

OBJECTIVE

This study examined quantitative heel ultrasonography (QUS), 25-hydroxyvitamin D (25-OHD) levels, and urine amino-terminal cross-linking telopeptide of type I collagen (NTX-I) levels in patients with a recent osteoporotic hip fracture to see whether they were clinically useful.

METHODS

Stiffness index (SI) T scores from QUS, 25-OHD levels, and urine NTX-I levels were obtained in 53 female and 32 male patients with hip fractures. Sixty-five female patients and 5 male patients attending our geriatric clinic were used for comparison.

RESULTS

The SI T scores of the hip fracture patients were less than those of the geriatric clinic patients. The difference was significant in female patients (P= .0001) but not in male patients (P= .1). Serum levels of 25-OHD were less than 28 ng/mL in 50 of 59 patients and less than 5 ng/mL in 2 patients. Levels of urine NTX-I were variable and were not correlated with other parameters.

CONCLUSIONS

Patients who have had a hip fracture have a low SI determined by QUS; this is easy to perform, and it provides a baseline T score from which to assess treatment effects. Most of these patients are vitamin D deficient, and measurement of the 25-OHD level would enable physicians to prescribe an appropriate dose of vitamin D. Urine NTX-I measured shortly after a hip fracture is not clinically helpful.

Aims: Recently, the determination of biochemical markers has been intensely explored to better understand the mechanisms underlying knee OA. In this study, we aimed to explore the expression pattern of five biochemical markers in patients with knee OA.

Methods: After IRB approval and signed informed consent, 26 patients were enrolled. Serum and synovial samples were collected prior to knee arthroscopy. Pre-operative assessment included diagnosis, Lysholm, Tegner Activity Scale, IKDC score, and radiographic Kellgren and Lawrence classification. ELISA of CTX-I, CTX-II, NTX-I, MMP3, and MMP13 were measured in serum and synovial fluid samples.

Results: Twenty-six patients were included, with a mean age of 42 ± 15 years old. Mean results and standard deviation of the biomarkers in serum were as follows: CTX-I 5.8 ± 5.5 ng/mL, CTX-II 3.8 ± 1.7 ng/mL, NTX-I 52 ± 71 (nM BCE), MMP3 1.18 ± 0.6 ng/mL, and MMP13 1243.6 ± 1422 pg/mL; synovial fluid results were as follows: CTX-I 0.74 ± 0.5 ng/mL, CTX-II 5.1 ± 2.5 ng/mL, NTX-I 254 ± 85 (nM BCE), MMP3 0.4 ± 0.4 ng/mL, and MMP13 797 ± 1391 pg/mL. We observed a differential pattern of expression in serum NTX-I in patients with chronic meniscus injuries when compared with ACL injuries or cartilage lesions.

Conclusions: In conclusion, the clinical criteria of early OA are useful to categorize patients with knee conditions. The biochemical markers explored did not yield a differential pattern that can be associated with this classification. Serum NTX-I could be a useful marker of chronic meniscal lesion in future longitudinal studies, after adjusting for age and sex.

Keywords: Biomarkers; Clinical criteria; Knee osteoarthritis.

Evidence suggests that endurance and even recreational cycling may stimulate bone resorption; however, little is known about cartilage response to endurance cycling exercise. We investigated effort-dependent changes in bone turnover and cartilage biomarkers in blood and urine samples from elite cyclists during a 3-week stage race. Whole blood and urine samples were collected the day before the start of the race, at mid and end-race for serum and urinary CTx-I, NTx-I, PINP, COMP (only in serum), and CTx-II analysis by enzyme-linked immunosorbent assay. The values were corrected for plasma volume or creatinine excretion, respectively, and correlated with power output (corrected for body weight) and net energy expenditure. Bone marker concentrations in both serum and urine were slightly but significantly decreased. Among the cartilage degradation markers, only CTx-II was decreased, while COMP remained unchanged. The changes in bone and cartilage turnover indexes were correlated with the indexes of physical effort and energy consumption. Strenuous physical effort, in the absence of mechanical loading, slows bone metabolism and only minimally affects cartilage turnover. Since changes in plasma and urine volume, which normally occur in exercising athletes, can mask these effects, biomarker concentrations need to be corrected for shifts in plasma volume and urinary creatinine for correct interpretation of the data.

Current knowledge about an incidence of bone metastases, use of bisphosphonates and assessment of response to the treatment are surveyed. The bone metastases are quite frequent in patients with breast and prostate cancers. High doses of intravenous pamidronate are particularly useful in the treatment of these patients. Prior to therapy with bisphosphonate special score elaborated by RE Coleman should be calculated. The new biochemical bone resorption markers especially Ntx i Crosslaps seems to be the most efficient for evaluation metastases response.

OBJECTIVE

To investigate the influencing factors and pathogenesis of osteopenia in the patients with hemophilia.

METHODS

Twenty-three patients with hemophilia were admitted in the hospital affiliated to North China University of Science and technology from March to August 2015, including 13 severe cases, 10 mild and moderate cases. All the patients accepted the detection of serum I collagen cross-linking N terminal peptide (NTX I), osteoprotegerin (OPG), bone alkaline phosphatase (BALP), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF) and transforming growth factor-β1 (TGF-β1), the score scale of activity ability was recorded according to the criteria published by the U.S. Center for disease prevention and control in 2002, and 21 patients received the measurement of bone mineral density. According to the World Health Organization (WHO) definition, the clinical significance of bone mineral density (BMD) was assessed by measuring the Z level.

RESULTS

Z level>-2 was recorded in 10 cases, Z≤-2 was recorded in 11 cases; the levels of body mass index (BMI) and human bone alkaline phosphatase (BALP) reflecting bone formation in 11 cases (Z≤-2) were lower than there in 10 cases (Z>-2) (P<0.05); the levels of BALP (r=0.489, P<0.05), IGF (r=0.538, P<0.05) and BMI (r=0.572, P<0.01) positively correlated significantly with BMD (P<0.05); the levels of bFGF (r=0.570, P<0.01) and OPG (r=0.505, P<0.05) positively correlated with NTX I, indicating bone destruction (P<0.05); the score of activity ability of severe patients was significantly lower than that of mild and moderate cases (P<0.05), BMD levels of these 2 groups were not statistically different (P>0.05).

CONCLUSIONS

The BMD level does not correlate with the clinial grouping of hemophilia, the low body mass index may be a risk factor for bone lose; the mechanism of hemophilia patient's bone lose may be related with the decrease of osteogenic activity, the IGF can prevent bone lose in hemophilia, the bFGF and OPG can promote bone metabolism of the patients with hemophilia.

BACKGROUND

The skeletal effects of low-dose methotrexate (MTX), in glucocorticoid-dependent asthmatics (GCDA), are unknown.

METHODS

We studied 9 patients from a total of 26 chronic GCDA who completed 28 weeks of MTX (15 mg weekly, intramuscularly). Prednisolone dose was not altered during the first 12 weeks, and was then reduced between 12 and 28 weeks. Mean (S.E.M.) age of the patients was 54 (4.0) years. They had normal bone mineral density (BMD), were not taking medication that affected bone metabolism (except prednisolone and inhaled corticosteroids) and all achieved at least 50% reduction in prednisolone dose at 28 weeks. Blood and urine samples were obtained at baseline, 12, 28 and 40 weeks for measurement of serum osteocalcin (OC) and bone alkaline phosphatase (Bone-ALP) as formation markers and urinary deoxypyridinoline (DPD) and N-terminal cross-linked telopeptide of type I collagen (NTX-I) as resorption markers.

RESULTS

Concurrently with the changes in prednisolone dosage serum OC levels increased significantly at 28 weeks (p<0.008) (8.1+/-1.0 ng/ml) compared to baseline (4.7+/-0.6 ng/ml) and 12 weeks (5.1+/-0.6 ng/ml), but trended back by 40 weeks (6.6+/-0.6 ng/ml). No significant changes were observed for the other bone markers between baseline and the other time points.

CONCLUSIONS

The beneficial effects of steroid reduction on bone metabolism do not appear to be impaired by concomitant MTX treatment at least over 12 weeks.

Introduction: Prolonged immobilization associated with several neurological disorders causes se condary osteoporosis with pathological fractures and persistent bone pain.

Objectives: To establish the association between bone mineral density (BMD), neoformation and bone resorption markers and the degree of functional capacity in children under 18 years of age with reduced mobility. Pa tients and Method: Cross-sectional study conducted in Ciudad Real, Spain between January 1, 2016, and December 31, 2017 with patients aged between 6 and 18 years diagnosed with different neuro logical disorders. The following variables were analyzed: age, sex, pubertal stage, functional capacity according to the Functional Mobility Scale (FMS), which assesses the ability to walk from 5, 50 to 500 meters, BMD, 25-hydroxy-vitamin D, alkaline phosphatase and osteocalcin in blood, and N-terminal telopeptide crosslinks in collagen type I (NTX-I) in urine. BMD, alkaline phosphatase, osteocalcin, and NTX-I values are expressed in Z score according to reference values for age and sex. The Pear son and Spearman correlations were used for data analysis.

Results: 36 patients (52.7% girls) with an average age of 8.6±4.7 years. Mean FMS value: 5.3 out of 18. Mean BMD: -1.99 ± 1.7 standard deviations (SD), mean alkaline phosphatase: -2.64 ± 1.08, mean osteocalcin: -2.15 ± 1.39, and mean NTX-I: +3 ± 1.72. There was a significant association between BMD and FMS for 5 meters (r = 0.395; p = 0.017) and for total score (r = 0.365; p = 0.029). There were no significant differences according to the stages of pubertal development.

Conclusions: In this population, there was a decrease in BMD and bone neoformation markers, and an increase of bone resorption markers with no association with pubertal development. Patients with a lower degree of mobility present a lower BMD.

The structure of the alpha-carbon chain was solved by molecular replacement method at 2.7 A resolution. Neurotoxin I (NTX-I) is one of the main protein components purified from the venom of the central asian cobra Naja naja oxiana. NTX-I is known to bind specifically to acetylcholine receptors thus preventing the transmission of the neuroconductivity signal from synapses to muscles. NTX-I crystals were grown either by vapour diffusion or dialysis methods using specially prepared microdialysis cell. The intensities of reflections from native NTX-I crystals were measured in the range of 38.0-2.1 A-1 by omega-scan method with a Syntex P21 diffractometer operated in automatic regime. To determine the position and mode of packing of NTX-I molecule in unit cell program packages MERLOT and BLANC were applied running on a NORD-500 computer.

Objectives: In this study, we aimed to investigate the bone turnover marker levels according to bisphosphonate usage and radiotherapy (RT) in cancer patients with metastases in osteolytic pattern.

Patients and methods: A total of 52 patients (13 males, 39 females; median age: 52 years; range, 37 to 78 years) treated with RT for osteolytic bone metastases between April 2005 and April 2006 were retrospectively analyzed. Bone-specific alkaline phosphatase (BAP), amino-terminal cross-linked telopeptide of type I collagen (NTX-I), amino-terminal propeptide of type I procollagen (PINP), osteocalcin (OC), deoxypyridinoline (DPD), pyridinoline (PYD), alkaline phosphatase (ALP), creatinine, calcium (Ca), phosphate (P), magnesium (Mg), and 24-h urine Ca levels were measured in blood and urine before the initiation of RT, six weeks and six months after RT.

Results: A decrease in BAP, PINP, and creatinine levels was observed after RT (Week 6 p=0.006, Month 6 p=0.008). Sixteen patients who already used bisphosphonate before RT were excluded from statistical calculation. The remaining 36 patients who were treated with bisphosphonate after the first blood test were evaluated separately. In this group of patients, BAP, PINP, NTX, creatinine, and Ca levels significantly increased at six weeks after RT. The PINP and creatinine values significantly decreased at six months after RT. The variation between two different RT arms was assessed with repeated measures variance analysis. There was a statistically significant difference for NTX, OC, and creatinine levels between the first and second measurements.

Conclusion: Radiotherapy is an effective method in the treatment of osteolytic bone metastases. Bone turnover markers can provide an objective evaluation on RT response and parallel to imaging modalities criteria for evaluation. Bisphosphonates may alter the levels of these indicators. However, in this study, there were no statistically significant differences between the levels of markers for two different RT schedules.

The aim of this study was to evaluate the effect of anti-tumor necrosis factor α (anti-TNF-α) therapy in combination with methotrexate on bone remodeling and osteoclastogenesis in female patients with RA. Serum levels of bone turnover markers (i.e., C- and N-terminal propeptides of type I procollagen (PICP and PINP), C- and N-terminal cross-linking telopeptides of type I collagen (CTX-I and NTX-I), and soluble receptor activator of nuclear factor κB ligand (sRANKL) and osteoprotegerin (OPG)) were determined by immunoassay at baseline and 15 months after initiation of treatment. Bone mineral density was measured by dual-energy x-ray absorptiometry. We found a significant decrease in serum PINP levels, a biomarker of bone formation, and higher levels of CTX-I and sRANKL indicative of increased bone resorption in RA patients prior to TNFαI treatment compared to the controls. Anti-TNF-α therapy was effective in improving bone metabolism in RA patients as reflected in a decrease in CTX-I (at least partially due to the RANKL/OPG reduction) and a concomitant increase in PINP levels. The bone metabolism changes were independent of the type of TNFαI used. PINP and CTX-I were found to be useful markers of bone metabolism, which may prove the effectiveness of TNF-α therapy earlier than the bone density assessment.

Keywords: CTX-I; NTX-I; PICP; PINP; RANKL/OPG; TNF-α inhibitors; bone turnover markers; rheumatoid arthritis.

Background: Extended-release naltrexone (XR-NTX), an opioid antagonist, has demonstrated equal treatment outcomes, in terms of safety, opioid use, and retention, to the recommended OMT medication buprenorphine. However, premature discontinuation of XR-NTX treatment is still common and poorly understood. Research on patient experiences of XR-NTX treatment is limited. We sought to explore participants' experiences with discontinuation of treatment with XR-NTX, particularly motivation for XR-NTX, experiences of initiation and treatment, and rationale for leaving treatment.

Methods: We conducted qualitative, semi-structured interviews with participants from a clinical trial of XR-NTX. The study participants (N = 13) included seven women and six men with opioid dependence, who had received a minimum of one and maximum of four injections of XR-NTX. The study team analyzed transcribed interviews, employing thematic analysis with a critical realist approach.

Findings: The research team identified three themes, and we present them as a chronological narrative: theme 1: Entering treatment - I thought I knew what I was going into; theme 2: Life with XR-NTX - I had something in me that I didn't want; and theme 3: Leaving treatment - I want to go somewhere in life. Patients' unfulfilled expectations of how XR-NTX would lead to a better life were central to decisions about discontinuation, including unexpected physical, emotional, or mental reactions as well as a lack of expected effects, notably some described an opioid effect from buprenorphine. A few participants ended treatment because they had reached their treatment goal, but most expressed disappointment about not achieving this goal. Some also expressed renewed acceptance of OMT. The participants' motivation for abstinence from illegal substances generally remained.

Conclusion: Our findings emphasize that a dynamic understanding of discontinuation of treatment is necessary to achieve a long-term approach to recovery: the field should understand discontinuation as a feature of typical treatment trajectories, and discontinuation can be followed by re-initiation of treatment.

Keywords: Extended-release naltrexone; Opioid use disorder; Patient experience; Treatment discontinuation.

There is an increasing incidence of destructive bone disease caused by osteoclast proliferation. This is characterized by reduced bone mass and imbalance of bone homeostasis. Icariin (ICA), a flavonoid compound isolated from Epimedium, has anti‑osteoporosis activity and inhibits the formation of osteoclasts and bone resorption. The purpose of the present study was to investigate the protective effect of ICA on osteoclastic differentiation induced by thioacetamide (TAA) and its possible mechanism in Sprague Dawley (SD) rats. In the present study, SD rats were intraperitoneally injected with TAA (300 mg/kg) for the bone loss model, treated with ICA (600 mg/kg, intragastric gavage) in the ICA group and TAA+ICA group for treatment of bone loss for 6 weeks. Indexes associated with bone metabolism, such as alkaline phosphatase, N‑terminal telopeptide of type‑I collagen (NTX‑I), calcium (Ca), phosphorus (P) and magnesium (Mg) in the serum, were detected. Osteoclast differentiation of femoral tissues was detected by hematoxylin and eosin and tartrate‑resistant acid phosphatase staining. The femoral bone mass was evaluated using a three‑point bending test and micro computed tomography. Western blotting was used to detect the expression levels of osteoclast‑related proteins in each group. In the rats treated with TAA, the serum concentrations of Ca, P and Mg were decreased, the serum concentration of NTX‑I was increased, osteoclast differentiation of the femur was increased, femur bone stress and bone mass were decreased and the bone loss and osteoclast formation were reduced after ICA treatment. In addition, ICA inhibited the protein expression of receptor activator of nuclear factor κ‑Β ligand (RANKL), receptor activator of nuclear factor κ‑B (RANK), p38, ERK, c‑Fos and nuclear factor of activated T cells 1 (NFATc1) in the femur of rats treated with TAA. The results suggested that ICA may inhibit osteoclast differentiation by downregulating the RANKL‑p38/ERK‑NFAT signaling pathway and prevent TAA‑induced bone loss. The results are helpful to understand the mechanism of osteoclast differentiation induced by TAA, as well as the antiresorptive activity and molecular mechanism of ICA, and to provide new ideas for the treatment of osteolytic diseases.

Keywords: bone loss; bone mineral density; icariin; nuclear factor of activated T cells; osteoclast; thioacetamide.

Background: An anterior cruciate ligament (ACL) rupture results in an increased risk of developing knee osteoarthritis (OA) at an early age. Before clinical signs become apparent, the OA process has already been initiated. Therefore, it is important to look at the cascade of changes, such as the activity of cytokines and proteases, which might be associated with the later development of OA.

Purpose: To compare biomarker levels in patients with a recent ACL rupture with those in controls with a healthy knee and to monitor biomarker levels over 2 years after an ACL rupture.

Study design: Descriptive laboratory study.

Methods: Patients were enrolled after an ACL tear was identified. Serum and urine samples were collected at the time of enrollment in the study (3-25 weeks after the injury) and then at 14 and 27 months after the injury between January 2009 and November 2010. Reference samples were obtained from participants with healthy knees. The following biomarkers were measured with immunological assays: aggrecan ARGS neoepitope (ARGS-aggrecan), tumor necrosis factor-α (TNF-α), interferon-γ, interleukin (IL)-8, IL-10, IL-13, N-terminal cross-linked telopeptide of type I collagen (NTX-I), and C-terminal cross-linked telopeptide of type II collagen (CTX-II).

Results: Samples were collected from 152 patients with an acute ACL rupture, who had a median age of 25 years (interquartile range [IQR], 21-32 years). There were 62 urine reference samples (median age, 25 years [IQR, 22-36 years]) and 26 serum reference samples (median age, 35 years [IQR, 24-39 years]). At a median of 11 weeks (IQR, 7-17 weeks) after trauma, serum levels of both ARGS-aggrecan and TNF-α were elevated 1.5-fold (P < .001) compared with reference samples and showed a time-dependent decrease during follow-up. Urine NTX-I and CTX-II concentrations were elevated in an early phase after trauma (1.3-fold [P < .001] and 3.7-fold [P < .001], respectively) compared with reference samples, and CTX-II levels remained elevated compared with reference samples at 2-year follow-up. Strong correlations were found between serum ARGS-aggrecan, urinary NTX-I, and urinary CTX-II (r_s = 0.57-0.68).

Conclusion: In the first few months after an ACL injury, there was a measurable increase in serum levels of ARGS-aggrecan and TNF-α as well as urine levels of NTX-I and CTX-II. These markers remained high compared with those of controls with healthy knees at 2-year follow-up.

Keywords: ACL injury; CTX-II; NTX-I; TNF-α; aggrecan; biomarkers.

(<em>i</em>)Background and Object<em>i</em>ves</<em>i</em>): The a<em>i</em>m of th<em>i</em>s study was to determ<em>i</em>ne the <em>i</em>nfluence of bone turnover markers, namely the N-term<em>i</em>nal cross-l<em>i</em>nk<em>i</em>ng telopept<em>i</em>de (<em>NTx</em>) and alpha C-term<em>i</em>nal cross-l<em>i</em>nk<em>i</em>ng telopept<em>i</em>de of type I collagen (α-CTx), <em>i</em>n detect<em>i</em>ng bone metastas<em>i</em>s (bone-only) <em>i</em>n breast cancer (BC) pat<em>i</em>ents, as well as to determ<em>i</em>ne whether th<em>i</em>s effect <em>i</em>s related to changes <em>i</em>n bone m<em>i</em>neral dens<em>i</em>ty (BMD). (<em>i</em>)Mater<em>i</em>als and Methods</<em>i</em>): The part<em>i</em>c<em>i</em>pants <em>i</em>n th<em>i</em>s study compr<em>i</em>sed 30 postmenopausal BC pat<em>i</em>ents w<em>i</em>th bone metastases (age range: 59.56 ± 9.02), 20 postmenopausal BC pat<em>i</em>ents w<em>i</em>thout bone metastases (age range: 55.30 ± 11.55), and 20 healthy postmenopausal female controls (age range: 55.55 ± 5.85). Bone turnover markers (serum <em>NTx</em> and ur<em>i</em>ne α-CTx) were measured us<em>i</em>ng the ELISA method. A dens<em>i</em>tometer us<em>i</em>ng dual-energy X-ray absorpt<em>i</em>ometry (DEXA) was used to analyze the BMD, and tumor markers were measured us<em>i</em>ng the chem<em>i</em>lum<em>i</em>nescent <em>i</em>mmunometr<em>i</em>c assay. (<em>i</em>)Results</<em>i</em>): The correspond<em>i</em>ng levels of serum <em>NTx</em> ((<em>i</em>)p</<em>i</em>) = 0.004), parathyro<em>i</em>d hormone (PTH) ((<em>i</em>)p</<em>i</em>) = 0.001), and ur<em>i</em>ne α-CTx ((<em>i</em>)p</<em>i</em>) < 0.001) of BC pat<em>i</em>ents were found to be h<em>i</em>gher than the standard levels. After the BC pat<em>i</em>ents were d<em>i</em>v<em>i</em>ded <em>i</em>nto subgroups on the bas<em>i</em>s of the presence of metastas<em>i</em>s, the ur<em>i</em>ne α-CTx levels ((<em>i</em>)p</<em>i</em>) = 0.001) were seen to be at cr<em>i</em>t<em>i</em>cally h<em>i</em>gh levels <em>i</em>n those pat<em>i</em>ents suffer<em>i</em>ng from BC w<em>i</em>th metastas<em>i</em>s. Though the BMD values <em>i</em>n the lumbar sp<em>i</em>ne ((<em>i</em>)p</<em>i</em>) < 0.001) and femoral neck ((<em>i</em>)p</<em>i</em>) = 0.001) were found to be s<em>i</em>gn<em>i</em>f<em>i</em>cantly low <em>i</em>n BC pat<em>i</em>ents, no stat<em>i</em>st<em>i</em>cally substant<em>i</em>al d<em>i</em>fference <em>i</em>n the BMD levels of BC pat<em>i</em>ents suffer<em>i</em>ng from metastas<em>i</em>s was observed. It was observed that ur<em>i</em>ne α-CTx (spec<em>i</em>f<em>i</em>c<em>i</em>ty: 70%; sens<em>i</em>t<em>i</em>v<em>i</em>ty: 85%) values are cr<em>i</em>t<em>i</em>cal factors that d<em>i</em>fferent<em>i</em>ate BC pat<em>i</em>ents w<em>i</em>th metastas<em>i</em>s from BC pat<em>i</em>ents w<em>i</em>thout metastas<em>i</em>s. (<em>i</em>)Conclus<em>i</em>ons</<em>i</em>): We found that alterat<em>i</em>ons <em>i</em>n bone turnover could be detected by us<em>i</em>ng the values of ur<em>i</em>ne α-CTx wh<em>i</em>le d<em>i</em>fferent<em>i</em>at<em>i</em>ng BC pat<em>i</em>ents w<em>i</em>th metastas<em>i</em>s from BC pat<em>i</em>ents w<em>i</em>thout metastas<em>i</em>s. Us<em>i</em>ng the b<em>i</em>ochem<em>i</em>cal markers of bone turnover and BMD together would be pert<em>i</em>nent for determ<em>i</em>n<em>i</em>ng the level of metastas<em>i</em>s present and exam<em>i</em>n<em>i</em>ng the eff<em>i</em>c<em>i</em>ency of bone dens<em>i</em>ty preservat<em>i</em>on therapy. Ideally, BMD measurement would be evaluated together w<em>i</em>th b<em>i</em>ochem<em>i</em>cal markers.

Keywords: C-terminal cross-linking telopeptide of type I collagen; N-terminal cross-linking telopeptide; bone mineral density; bone turnover markers; breast cancer.