Activin Research Articles

СИСТЕМА ФОРМИРОВАНИЯ ЦЕЛЕВОЙ КАРТИНЫ ДЕЯТЕЛЬНОСТИ ПРОИЗВОДСТВЕННОЙ СИСТЕМЫ В ДИНАМИЧЕСКИ ИЗМЕНЯЮЩИХСЯ УСЛОВИЯХ

СИСТЕМА ФОРМИРОВАНИЯ ЦЕЛЕВОЙ КАРТИНЫ ДЕЯТЕЛЬНОСТИ ПРОИЗВОДСТВЕННОЙ СИСТЕМЫ В ДИНАМИЧЕСКИ ИЗМЕНЯЮЩИХСЯ УСЛОВИЯХ

Hypoxia enhances differentiation of mouse embryonic stem cells into definitive endoderm and distal lung cells.

We investigated the effects of hypoxia on spontaneous (SP)- and activin A (AA)-induced definitive endoderm (DE) differentiation of mouse embryonic stem cells (mESCs) and their subsequent differentiation into distal pulmonary epithelial cells. SP differentiation for 6 days of mESCs toward endoderm at hypoxia of 1% O2, but not at 3% or 21% (normoxia), increased the expression of Sox17 and Foxa2 by 31- and 63-fold above maintenance culture, respectively. Treatment of mESCs with 20 ng/mL AA for 6 days under hypoxia further increased the expression of DE marker genes Sox17, Foxa2, and Cxcr4 by 501-, 1,483-, and 126-fold above maintenance cultures, respectively. Transient exposure to hypoxia, as short as 24 h, was sufficient to enhance AA-induced endoderm formation. The involvement of hypoxia-inducible factor (HIF)-1α and reactive oxygen species (ROS) in the AA-induced endoderm enrichment was assessed using HIF-1α(-/-) mESCs and the ROS scavenger N-acetylcysteine (NAC). Under SP conditions, HIF-1α(-/-) mESCs failed to increase the expression of endodermal marker genes but rather shifted toward ectoderm. Hypoxia induced only a marginal potentiation of AA-induced endoderm differentiation in HIF-1α(-/-) mESCs. Treatment of mESCs with AA and NAC led to a dose-dependent decrease in Sox17 and Foxa2 expression. In addition, the duration of exposure to hypoxia in the course of a recently reported lung differentiation protocol resulted in differentially enhanced expression of distal lung epithelial cell marker genes aquaporin 5 (Aqp5), surfactant protein C (Sftpc), and secretoglobin 1a1 (Scgb1a1) for alveolar epithelium type I, type II, and club cells, respectively. Our study is the first to show the effects of in vitro hypoxia on efficient formation of DE and lung lineages. We suggest that the extent of hypoxia and careful timing may be important components of in vitro differentiation bioprocesses for the differential generation of distal lung epithelial cells from pluripotent progenitors.

P36. The prognostic potential of circulating and tissue activin A level in malignant pleural mesothelioma.

Background Malignant pleural mesothelioma (MPM) is an aggressive malignancy characterized by poor outcome and there is a lack of prognostic biomarkers to estimate prognosis. Previously we have shown that suppression of activin A can interfere with MPM tumor growth. In the current study, we compare the circulating and tissue expression level of activin A as a prognostic biomarker in MPM.

Short call abstracts.

Short call abstracts.

Association of serum bisphenol-A concentration and male reproductive function among exposed workers.

Bisphenol-A (BPA) can alter endocrine function in animals, yet the relationship between human exposure to BPA and male reproductive function is not well understood. We collected serum samples from 281 male workers exposed to BPA and 278 controls. Serum BPA concentrations were analyzed by gas chromatography-mass spectrometry after derivatization. The serum levels of sex hormone-binding globulin (SHBG) and total testosterone (TT) were detected by radioimmunoassay and levels of inhibin B (INB) and androstenedione (AD) by enzyme-linked immunosorbent assay. We examined the association of BPA exposure and male sex hormone levels by multivariable linear regression. Both serum BPA concentrations and detection rates were greater in males exposed than those not exposed to BPA, but the two groups did not differ in levels of SHBG, TT, INB, or AD. Compared with exposure for ≤5 years, exposure for >5 years was associated with increased serum BPA and SHBG 1evels and decreased AD levels. Compared with undetectable BPA and BPA ≤ 18.75 ng/mL, BPA level >18.75 ng/mL was associated with low AD and high SHBG levels (P < 0.05). On adjusted multivariable regression, increased serum BPA level was associated with decreased mean serum AD level (0.18 ng/mL; 95% confidence interval CI -0.22 to -0.13) and increased mean serum SHBG level (2.79 nmol/L; 95% CI 2.11-3.46). Serum BPA levels were increased after occupational exposure. BPA exposure was negatively associated with serum AD level but positively associated with serum SHBG level.

Involvement of Human Seminal Plasma Inhibin, a Multifunctional Protein, in Anti-HIV Activity

Body fluids of mammals, including semen, are increasingly recognized to harbour anti-microbial proteins which play a role in host defence against a myriad of pathogens. Human seminal plasma inhibin (hSPI) is reported to be a multifunctional protein, well-studied primarily for its fertility-related effects and recently for its anti-fungal activity. Therefore, it was thought worthwhile to study whether it also possesses anti-HIV activity. Towards this objective, human seminal plasma proteins were fractionated using gel-permeation chromatography and chromatofocusing. Results demonstrated that fraction-5 (containing purified hSPI) of the chromatofocused fractions demonstrated substantial anti-HIV activity. The data suggest that hSPI possesses anti-HIV activity. In future, molecular models based on such naturally occurring proteins could be potentially employed for prevention/ therapeutics for HIV/AIDS.

Prevalence of thyroid disorders and the correlation of thyroid profile with liver enzymes, serum activin-A and follistatin during the treatment of patients with chronic hepatitis C genotype 1 and 4

Amac: Kronik hepatit C (KHC) ve Peg-interferon- alfa (Peg-INF-α) serum aktivinleri ve follistatini duzenler ve tiroid hastaliklari (TH) ile birliktedir. Bu calismanin amaci KHC\'nin indukledigi TH sikligini belirlemek ve karaciger hasari, serum activin-A ve follistatin\'in tiroid fonksiyonlari ve tiroid otoantikorlari ile iliskisini arastirmaktir. Yontemler: Bu calisma kesitsel olup, 3 gruba ayrilan 132 KHC\'li hastadan serumlari alinmistir: Tedavisiz 56 hasta, 24 haftalik Peg-INF-α tedavisi almis 30 hasta ve Peg-INF-α tedavisi tamamlanmis 46 hasta. Tiroid stimulan hormon (TSH), serbest tiroksin (FT4) ve tiroid otoantikorlari, serum activin-A ve follistatin duzeyleri ELISA yontemiyle olculdu. Bulgular: Hastalarin %15\'inde (n=20) tiroid hastaligi saptandi, bunlarin cogunlugu (%80) otoimmun tiroidit olup, kadinlarda daha sik (%70) idi. TSH reseptor antikorlari (TSHR-Abs), diger antikorlardan anlamli yuksek bulundu (p

HUBUNGAN AKTIVITAS FISIK DENGAN KEJADIAN OBESITAS PADA ORANG DEWASA DI INDONESIA (Analisis Data Riskesdas 2007)

ASSOCIATION BETWEEN PHYSICAL ACTIVITY AND ADULT OBESITY IN INDONESIA (Secondary Data Analysis of the Basic Health Research 2007) The enhancement of social economy status and the life style changes, including changes in eating habits and reduction of physical activities, increased overweight and obesity revalence. This paper aims to study the association between physical activity and adult obesity in Indonesia. This research used Basic Health Research (Riset Kesehatan Dasar) 2007 data with cross-sectional design. The populations were all household members aged ≥ 18 years, whereas the samples were all household members aged ≥ 18, physically and mentally healthy, non-pregnant, and their BMI were at least 18.5 kg/m 2. The respondent was obese if his/her BMI was at least 18.5 kg/m2. Physical activity was assessed with questions about its type (vigorous and moderate) and its duration per week. Logistic regression was used to study the relationship between physical activity and adult obesity. The result showed that Obesity prevalence (BMI&gt;27kg/m2) in adult was 12.47% (CI 95%: 12.28 – 12.66). The result showed tthere was a relationship between physical activity and adult obesity, controlled by confounder variable (area, age, sex, marital status, education, occupation, and smoking habits). People who had low level of physical activity had more risk to be obese than those who had high level physical activity. It has been recommended that to reduce the risk of being obese, it is advisable to have sufficient physical activity such as alking, jogging, running, and biking. It is also necessary for the Ministry of Health to develop guidance of sufficient physical activity. Keywords : obesity, adult, physical activity

RELATIONSHIP OF LIFE SPACE AND ACTIVITY IN LATE LIFE WITH NEUROPATHOLOGY

RELATIONSHIP OF LIFE SPACE AND ACTIVITY IN LATE LIFE WITH NEUROPATHOLOGY

Effects of tripterygium wilfordii polyglucoside on expression of Activin A and influence of transdifferentiation in renal tubulointerstitium of rats with diabetic nephropathy

To explore the effects of tripterygium wilfordii polyglucoside (TWP) on expression of Activin A (Act-A) and influence of transdifferentiation in renal tubulointerstitium of rats with diabetes nephropathy (DN). A total of 40 male Wistar rats were randomly divided into normal control, DN model, irbesartan treatment and TWP treatment groups (n = 10 each). The animal model of DN was established by an injection of streptozotocin. After modeling, irbesartan treatment and TWP treatment groups received an intragastric injection of corresponding drugs. The body weights were recorded and the 24 h Pro quantitative levels detected at Weeks 4, 8, 12 and 16 respectively. After sacrificing at Week 16, serum creatinine (Scr) was detected. The renal tissues were excised and examined after HE and PAS staining. The expressions of Act-A, alpha-smooth muscle actin (α-SMA), collagen type IV (Co-IV) and E-cad in renal tubulointerstitium were detected by immunohistochemistry. And the expressions of Act-A, α-SMA, Co-IV and mRNA of E-cad in renal tissue were detected by real-time quantitative fluorescent polymerase chain reaction (PCR). Compared with DN model group, 24 hPro ((67.1 ± 9.6), (61.8 ± 8.0) vs (158.3 ± 13.3) mg), quantitative Scr ((31.7 ± 4.1), (32.6 ± 6.2) vs (55.3 ± 8.9) µmol/L) and renal hypertrophic index ((9.14 ± 0.65), (7.50 ± 0.34) vs (11.28 ± 0.70) mg/g) of irbesartan treatment and TWP treatment groups decreased obviously at Week 16 (all P < 0.05). However, no statistical significance existed between irbesartan treatment and TWP treatment groups. The body weights of irbesartan treatment and TWP treatment groups increased versus DN model group. Pathological sections showed that lesion degree of renal tubulointerstitium became alleviated in each group. Yet no statistical significance existed between irbesartan treatment and TWP treatment groups. Both immunohistochemistry and real-time quantitative fluorescent PCR showed that the expressions of Act-A, α-SMA and Co-IV significantly decreased in renal tubulointerstitium (P < 0.01) while the expression E-cad significantly increased (P < 0.01) in inirbesartan treatment and TWP treatment groups versus DN model group. And no statistical significance existed between irbesartan treatment and TWP treatment groups. TWP may down-regulate the expression of Act-A and inhibit the transdifferentiation of renal tubulointerstitial so as to delay renal fibrosis.

Long-term imatinib treatment does not cause testicular toxicity in male adolescents with chronic myeloid leukemia and in a juvenile rat model.

The impact of exposure to the tyrosine kinase inhibitor (TKI) imatinib (IMA) on the male reproductive endocrine system is still discussed controversially. We therefore investigated testosterone (Testo) and inhibin B (InB) in blood serum from male adolescents with chronic myeloid leukemia (CML) under long-term TKI treatment. Also long-term exposure to TKIs was studied in a juvenile rat model. Serum was collected at 3 months intervals from 13 boys (age: 7.8-18.9 years, median: 12.8 years) with CML receiving TKI treatment over 3-58 months (median: 18 months). 4 weeks (w) old male rats were exposed, either chronically or intermittently, via the drinking water to a standard (SD) and a high dose (=2-fold SD) of IMA, dasatinib (DASA), or bosutinib (BOSU) over a 10 w period. Controls received water only. Animals were sacrificed after 2 w (prepubertal), 4 w (pubertal), and 10 w (postpubertal) of exposure. Testo and InB serum levels were measured by ELISA. Boys exhibited Testo and InB levels within normal age-related reference ranges and no pattern of rising or falling levels during TKI treatment could be observed. In rats, Testo levels under IMA exposure tended to be non-significantly lowered at postpubertal age compared to controls while no significant differences were found under DASA and BOSU exposure. Animals' InB levels did not significantly differ from controls for all TKIs, at all doses, and by all application schemes tested. With the limitation that the number of individuals tested was rather small, testicular toxicity due to TKI seems unlikely as no alterations of Testo and InB blood levels neither in male adolescent patients nor in rats under long-term TKI exposure was observed.

Toxic Effects of Atrazine on Reproductive System of Male Rats

ObjectiveThis study was designed to evaluate the toxic effects of Atrazine (ATZ) on the reproductive system of male rats. MethodsMale Sprague-Dawley rats were exposed to ATZ by gavage at dosages of 0, 38.5, 77, and 154 mg/kg bw/day for 30 d. The toxic effects of ATZ to rats were assessed through histopathologcal observation, spermatozoa quality evaluation, testicular marker enzyme indicators, antioxidant capacity and reproductive hormone levels. ResultsSignificant adverse effects on reproductive system were observed in rats exposed to ATZ at different dosages compared with 0 mg/kg group, including an irregular and disordered arrangement of the seminiferous epithelium in 154 mg/kg group; a decreased spermatozoa number and an increased spermatozoa abnormality rate in 77 and 154 mg/kg groups; decreased levels of acid phosphatase (ACP), alkaline phosphatase (AKP), lactic dehydrogenase (LDH), and succinate dehydrogenase (SDH) with the increasing of ATZ concentration; a decreased level of total antioxidant capacity (TAC) in a dose-dependent manner, and a decreased reduced glutathione (GSH) level and an increased malondialdehyde (MDA) content in 154 mg/kg group; and decreased serum levels of testosterone (T) and inhibin-B (INH-B) and an increased serum level of follicle stimulating hormone (FSH) in 77 and 154 mg/kg groups, and an increased serum level of luteinizing hormone (LH) in 154 mg/kg group. ConclusionsThese results suggested that relatively high doses of ATZ could exert reproductive toxicity of male rats.

Gonadal function in males with autoimmune Addison's disease and autoantibodies to steroidogenic enzymes

Steroidogenic enzyme autoantibodies (SEAbs) are frequently present and are markers of autoimmune premature ovarian failure (POF) in females with autoimmune Addison's disease (AAD). The prevalence and significance of SEAbs in males with AAD have not yet been defined. We studied the prevalence of SEAbs in a large cohort of males with AAD and assessed the relationship between SEAbs positivity and testicular function. A total of 154 males with AAD (mean age 34 years) were studied. SEAbs included autoantibodies to steroid-producing cells (StCA), detected by immunofluorescence, and steroid 17α-hydroxylase (17α-OHAbs) and side chain cleavage enzyme (SCCAbs) measured by immunoprecipitation assays. Gonadal function was evaluated by measuring follicle-stimulating hormone (FSH), luteinizing hormone (LH), total testosterone (TT), sex hormone-binding globulin (SHGB), anti-müllerian hormone (AMH) and inhibin-B (I-B). Twenty-six males, 10 SEAbs((+)) and 16 SEAbs((-)), were followed-up for a mean period of 7·6 years to assess the behaviour of SEAbs and testicular function. SEAbs were found in 24·7% of males with AAD, with the highest frequency in patients with autoimmune polyendocrine syndrome type 1 (APS-1). The levels of reproductive hormones in 30 SEAbs((+)) males were in the normal range according to age and were not significantly different compared to 55 SEAbs((-)) males (P > 0·05). During follow-up, both SEAbs((+)) and SEAbs((-)) patients maintained normal testicular function. SEAbs were found with high frequency in males with AAD; however, they were not associated with testicular failure. This study suggests that the diagnostic value of SEAbs in males with AAD differs compared to females, and this may be related to the immunoprivileged status of the testis.

MP17-01 ACID-SENSING ION CHANNELS ARE INVOLVED IN AFFERENT EXCITATORY TRANSMISSION CONTROLLING REFLEX BLADDER ACTIVITY IN MICE

You have accessJournal of UrologyBladder & Urethra: Anatomy, Physiology & Pharmacology1 Apr 2014MP17-01 ACID-SENSING ION CHANNELS ARE INVOLVED IN AFFERENT EXCITATORY TRANSMISSION CONTROLLING REFLEX BLADDER ACTIVITY IN MICE Mitsuharu Yoshiyama, Hideki Kobayashi, Isao Araki, and Masayuki Takeda Mitsuharu YoshiyamaMitsuharu Yoshiyama More articles by this author , Hideki KobayashiHideki Kobayashi More articles by this author , Isao ArakiIsao Araki More articles by this author , and Masayuki TakedaMasayuki Takeda More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.522AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Acid-sensing ion channels (ASICs) have been proposed as transducers of sensory stimuli. ASIC subunits mRNAs are largely expressed in the central nervous systems (CNS), the peripheral nervous system including dorsal root ganglia (DRG) that innervate the bladder, and the bladder (both mucosa and detrusor), suggesting the possibility that ASICs are involved in regulation of lower urinary tract (LUT) activity. In this study, we examined effects of A-317567, a non-amiloride blocker of ASICs, on activity of LUT with or without acid irritation. METHODS C57BL/6 female mice (12-13 week-old) were anesthetized with sevoflurane during surgery including precollicular decerebration. A PE-50 tube was inserted into the bladder dome to record intravesical pressure. Cystometrogram (CMG) recordings under unanesthetized conditions were performed by continuously infusing (30 μl/min) saline alone or saline followed by dilute acetic acid (A/A, pH 3). CMG parameters were: pressure threshold for inducing micturition (PT); maximal voiding pressure (MVP), bladder compliance (BCP), bladder contraction duration (BCD), and inter-contraction interval (ICI). A-317567 was infused intravesically (100 μM) or injected i.p. (30 mg/kg), and effects of the drug were compared with those of the vehicle. All values are expressed as mean ± S.E.M. For statistical analysis, two-way repeated measures ANOVA and Wilcoxon matched-pairs signed rank test were used, and P ≤ 0.05 was considered significant. RESULTS I.p. administration of A-317567 increased PT and ICI and decreased BCP during saline infusion CMG, whereas it produced no changes in MVP and BCD (Fig. 1A). The i.p. dose ameliorated bladder hyperactivity (i.e., exhibited as decreased ICI) induced by intravesical A/A perfusion (Fig. 1B). However, intravesical infusion of the drug had no effects on activity of the LUT either with or without acid irritation. Neither intravesical infusion nor i.p. injection of the vehicle produced significant effects on both normal LUT activity and A/A-induced bladder hyperactivity. CONCLUSIONS ASICs participate in afferent excitatory transmission controlling reflex bladder activity under conditions of the LUT either with or without acid irritation. ASICs in the DRG, the CNS or both, but not in the bladder, play the essential role. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e135-e136 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Mitsuharu Yoshiyama More articles by this author Hideki Kobayashi More articles by this author Isao Araki More articles by this author Masayuki Takeda More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

MP24-17 INTERLEUKIN-15 DOWNREGULATES INHIBITORY NATURAL KILLER(NK) CELL LIGANDS EXPRESSED ON PROSTATE CANCER CELLS RESULTING IN INCREASED NK CELL ACTIVITY IN PROSTATE CANCER-IMMUNE EFFECTOR CELL COCULTURES.

You have accessJournal of UrologyProstate Cancer: Basic Research I1 Apr 2014MP24-17 INTERLEUKIN-15 DOWNREGULATES INHIBITORY NATURAL KILLER(NK) CELL LIGANDS EXPRESSED ON PROSTATE CANCER CELLS RESULTING IN INCREASED NK CELL ACTIVITY IN PROSTATE CANCER-IMMUNE EFFECTOR CELL COCULTURES. Christine Galustian, Christina Sakellariou, Oussama Elhage, Osamu Ukimura, Inderbir Gill, Richard Smith, and Prokar Dasgupta Christine GalustianChristine Galustian More articles by this author , Christina SakellariouChristina Sakellariou More articles by this author , Oussama ElhageOussama Elhage More articles by this author , Osamu UkimuraOsamu Ukimura More articles by this author , Inderbir GillInderbir Gill More articles by this author , Richard SmithRichard Smith More articles by this author , and Prokar DasguptaProkar Dasgupta More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.298AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The prostate cancer microenvironment is profoundly immunosuppressive and NK cells and T cells that infiltrate this environment are rendered anergic or suppressive. Recently, immunomodulatory checkpoints such as PDL1 and CTLA-4 have been shown to be important in creating this immunosuppression. Antibodies to these have a moderate effect in suppressing tumour growth but many unknown immunosuppressive factors exist on tumour cells. We have shown that prostate cancer cells express a wide range of ligands that inhibit NK cell activity by binding to inhibitory Killer Ig like receptors (KIRs). These receptors are critical in modulating the function of NK cells. Also, these cells have a downregulated expression of HLA molecules that render them less susceptible to T cell mediated recognition and killing. We have shown that IL-15, unlike other immunotherapeutic cytokines (e.g IL-12, IL-21), increases NK and CD8-T-cell activity within the prostate cancer microenvironment. We have previously demonstrated that IL-15 upregulates activatory KIRs and downregulates inhibitory KIRs to increase NK cell activity. To further these investigations, we have now examined the effects of IL-15 on ligands on prostate cancer cells that are recognised by activatory and inhibitory KIRs. METHODS PC3 and LNCaP prostate cancer cell-lines were incubated with non-adherent PBMCs at effector:target ratios of 8:1 and cytokines IL-2 or IL-15. After 1 week, Tumour cells were stained with antibodies to inhibitory HLA-class1-BW4 and HLA-G, and activatory Nectin-2 and MICA/MICB KIR-ligands. Antibody to HLA-ABC was also used. Flow cytometric analysis followed staining. RESULTS IL-15, but not IL-2 decreased the inhibitory-ligands HLA-Class-1-BW4 by upto 60%* on both LNCaP and PC3 cells. IL-15 also decreased the expression of HLA-G by 75%* on LNCaP cells. HLA-A,B,C expression was increased by over 65%* in the two prostate cancer cell lines. Initial data showed that activatory-ligands were not affected on either PC3 or LNCaP tumour cells. * =(p<0.001,1-way-anova, post-hoc Newman-Keuls,n=5). CONCLUSIONS IL-15 can downregulate inhibitory NK ligands on prostate cancer cells in addition to its ability to modulate activatory and inhibitory KIRs. This suggests strong shifts toward NK activation vs inhibition in the cocultures. Increased HLA-A,B,C on the tumour cells also favours stronger cytotoxic-T cell recognition. The molecular basis of these effects are currently being investigated. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e265 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Christine Galustian More articles by this author Christina Sakellariou More articles by this author Oussama Elhage More articles by this author Osamu Ukimura More articles by this author Inderbir Gill More articles by this author Richard Smith More articles by this author Prokar Dasgupta More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Chip is a critical regulator of traf6 degradation and osteoclast formation

Chip is a critical regulator of traf6 degradation and osteoclast formation

Exploring a role for the serine protease PACE4 in pain associated with experimental OA

Exploring a role for the serine protease PACE4 in pain associated with experimental OA

Hydrostatic pressure induces increased production of TGF-β via ERK activation in synovial fibroblasts from rzat temporomandibular joint

Hydrostatic pressure induces increased production of TGF-β via ERK activation in synovial fibroblasts from rzat temporomandibular joint

Bone turnover markers (BTMs) in the synovial fluid in the patients with osteoarthritis correlates with the serum level of BTMs though some patients with osteoporosis exhibit mismatched increase in bone resorption activity in synovial fluid

s / Osteoarthritis and Cartilage 22 (2014) S57–S489 S375 663 BONE TURNOVER MARKERS (BTMS) IN THE SYNOVIAL FLUID IN THE PATIENTS WITH OSTEOARTHRITIS CORRELATES WITH THE SERUM LEVEL OF BTMS THOUGH SOME PATIENTS WITH OSTEOPOROSIS EXHIBIT MISMATCHED INCREASE IN BONE RESORPTION ACTIVITY IN SYNOVIAL FLUID Y. Saita yz, K. Kurata z, T. Sato z, S. Fukasaku z, K. Kaneko y, M. Nemoto z. yDept. of Orthopaedics, Juntendo Univ., Tokyo, Japan; zDept. of Orthopedic Surgery, Kitanarashino Hanawa Hosp., Chiba, Japan Purpose: Little is known regarding the relationships between the osteoarthritis (OA) and osteoporosis especially for the link between the subchondral bone remodeling and pathology of OA. In the patients with severe OA, articular cartilages wear down and subchondral bones become exposed. We hypothesized that BTMs in the synovial fluid would be related to the pathology of OA, and evaluated them in the patients with knee OA. Methods: Twenty seven knees in twenty one postmenopausal women with advanced-stage knee osteoarthritis (Kellgren-Lawrence >2) were analyzed in this study. The average age of patients was 73.4 (range, 52– 83 years). Knee pain was evaluated using visual analogue scales (VAS) and outcome measure for Japanese patients with knee osteoarthritis (JKOM). Tartrate-resistant acidic phosphatase 5b (TRAcP5b) activity and procollagen 1 N-terminal peptide (P1NP) concentrationwere measured both in the serum and synovial fluid. Bone mineral density (BMD) was measured in digital image processing method at second metacarpal bone. In statistics, Spearman’s rank correlation coefficient (r) and statistical significance (p) were calculated using GraphPad Prismversion 6.0 (GraphPad Software). The Mann-Whitney U test was used to compare differences between two independent groups. The values of p 0.5 mm in the index knee) at 3 years was obtained using a Poisson regression model with robust error estimates to obtain relative risk (RR), with adjustment for baseline index JSW, gender, glucosamine use, age, BMI and centre. Each knee was graded according to the Kellgren and Lawrence (KL placebo 0.08 mm/year; 95% CI 0.15–0.30, p 1⁄4 0.51). In the lateral compartment, the equivalent difference between the two groups was 0.07 mm/year (vitamin D 0.11mm/year, placebo 0.18mm/year; 95% CI 0.15–0.30). 39% of patients in the treatment group and 37% of patients in the placebo group showed a clinically significant progression of JSN (>0.5 mm) at 3 years compared to baseline. The unadjusted relative risk of clinically significant progression of JSN for the vitamin D group relative to the placebo group was 1.05 (95% CI 0.77–1.44, p 1⁄4 0.76). The adjusted relative risk was 1.03 (95% CI 0.76–1.40, p 1⁄4 0.83). The odds ratio of a higher KL OVX with alendronate (ALN,

Environmental pollutants and osteoarthritis: Effects of non-dioxin-like polychlorinated biphenyls on cultured chondrocytes

s / Osteoarthritis and Cartilage 22 (2014) S57–S489 S173 used in experiments up to the 2nd passage. Two experimental protocols were used: a membrane water permeability assay and measurement of intracellular calcium using Fura-2AM. Results: Using an established permeability assay, whereby a hypotonic solution is applied to the cells and cell volume recorded, aquaporin permeability was measured. In healthy cells permeability was found to be 31 3x104cm.s (n 1⁄4 5). Stimulated cells did not show a significant change in aquaporin permeability (n 1⁄4 12). Calcium measurements showed that healthy cells responded to the same hypotonic challenge with an intracellular calcium increase of 115 15nM (n 1⁄4 16). This calcium increase was inhibited by the TRP channel antagonist, PYR3 (n 1⁄4 69). When applied to cells from the in vitro model of arthritis the same osmotic challenge caused a significantly greater calcium increase of 328 45nM (n 1⁄4 11; p 0.01). Conclusions: We have investigated changes to two ion channels in healthy chondrocytes and those from an in vitro model of arthritis. Aquaporin function appeared unchanged in our cytokine arthritis model, possibly suggesting that this change in gene expression occurs as a result of OA, rather than contributing to OA development. Previous work has shown that intracellular calcium increases greater than :300nM can lead to cell apoptosis and therefore the changes we observe here could implicate important pathological changes in cytokinestimulated chondrocytes. Further work is necessary to identify the exact mechanism by which this calcium increase occurs. 291 ENVIRONMENTAL POLLUTANTS AND OSTEOARTHRITIS: EFFECTS OF NON-DIOXIN-LIKE POLYCHLORINATED BIPHENYLS ON CULTURED CHONDROCYTES V. Abella yz, M. Scotece y, J. Conde y, V. Lopez y, V. Lazzaro yx, J. Pino y, J.J. Gomez-Reino y, R. Meli k, O. Gualillo y. y SERGAS-IDIS Santiago Univ. Clinical Hosp., Santiago de Compostela, SPAIN; zUniv. of A Coru~ na, A Coru~ na, Spain; xUniversit a della Magna Grecia, Catanzaro, Italy; kUniv. of Naples Federico II, Napoli, Italy a. Purpose: Non-dioxin-like polychlorinated biphenyls (PCBs) are persistent organic pollutants that accumulate in fatty tissues causing immune suppression and endocrine disruption. Several studies suggest that PCBs may be involved in pathogenesis of osteoarthritis (OA). Alterations in the mechanisms of programmed cell death (apoptosis) are strongly related to the degradation of extracellular matrix (ECM) in the cartilage of OA subjects. Identification of apoptosis inducers is of paramount relevance to understand the pathogenesis and/or progression of OA. Thus, the aim of the present study was to assess the effect of several PCBs on chondrocytes viability and apoptosis induction. b. Methods: The murine chondrogenic cell line ATDC5 and human juvenile costal chondrocyte cell line T/C-28a2 were treated with several doses of PCB 101, 153 and 180, alone and in combination. Cell viability was examined using a colorimetric assay based on the MTT labeling reagent. Apoptosis was evaluated by Annexin V flow cytometric assay and by the involvement of apoptotic related proteins, such as caspase-3 and Bcl-2/Bax ratio, using western blot analysis. Finally, to evaluate whether PCBs exert necrotic effect, apart from apoptosis pathways, we have also assessed lactate dehydrogenase (LDH) levels in culture supernatants. c. Results: ATDC5 and T/C-28a2 cell lines treated with PCBs, alone and in combination, showed a significant reduction of cell viability rate in a concentration-dependent manner. Neither synergisms nor additive effects were observed on cell viability with the combined treatment. Data from annexin V assays suggested that PCBs clearly induced apoptotic pathways, as well as, a certain rate of necrosis. Actually, this effect was confirmed by evaluating LDH levels that were strongly increased in supernatants of PCBs-treated cells, suggesting that necrotic mechanisms are at play too. PCBs also induced caspase-3 activation by increasing its proteolytic cleavage in a concentration-dependent manner. Finally, the Bcl2/Bax ratio was also altered. d. Conclusions: The viability of murine and human chondrocytes was reduced in presence of PCBs. The activity of PCBs on cell viability is likely to be mediated by alterations in the mechanisms of regulation of apoptosis and necrosis. Overall, this work highlights a novel role of environmental pollutants in the pathophysiology of chondrocytes. 292 IL-1b MEDIATES MMP SECRETION AND IL-1B NEOSYNTHESIS VIA UPREGULATION OF P22PHOX AND NOX4 ACTIVITY IN HUMAN ARTICULAR CHONDROCYTES F. Rousset y, F. Hazane Puch z, L. Grange yz, M. Nguyen y, C. Pinosa y, C. Combaz Lair z, B. Burroni z, B. Rubens-Duval z, F. Morel y, B. Lardy yz. yUniversit e Joseph Fourier, Grenoble, France; zCHU Grenoble, Grenoble,