Ferric carboxymaltose therapy reduces pain and improves the quality of life in female patients with fibromyalgia.

OBJECTIVE: The efficacy of treatments for fibromyalgia is limited and many factors have been identified to trigger the current complaints. Iron deficiency anemia is one of these factors. We aimed to re-evaluate the quality of life of fibromyalgia patients with the Fibromyalgia Impact Questionnaire after ferric carboxymaltose treatment. PATIENTS AND METHODS: This study was conducted on 90 female patients older than 18 years of age with ferritin <60 mcg/dL who presented to the Internal Medicine outpatient clinic in a large tertiary care hospital in Eskisehir, Turkey, with FM symptoms. Patients were selected from women who had previously received oral iron therapy for at least 3 months and whose ferritin could not be increased to >60 mcg/dL. Patients who met the 2010 criteria of the American College of Rheumatology for fibromyalgia were included in the study. Patient characteristics and laboratory parameters were recorded. The Fibromyalgia Impact Questionnaire (FIQ1, FIQ2) was applied and compared before and after IV iron treatment. RESULTS: The mean age of the patients was 40±12 years (18-83). There was a significant change in the total score (FIQ1 mean: 54, FIQ2 mean: 21) and all parameters of the FIQ questionnaire after ferric carboxymaltose treatment (p=0.000). CONCLUSIONS: Ferric carboxymaltose treatment reduces pain levels and improves the quality of life in women with fibromyalgia.

Evaluation of circulating microRNA profiles in blood as potential candidate biomarkers in a subacute ruminal acidosis cow model - a pilot study.

BACKGROUND: Subacute ruminal acidosis (SARA) is a metabolic disorder often observed in high-yielding dairy cows, that are fed diets high in concentrates. We hypothesized that circulating miRNAs in blood of cows could serve as potential candidate biomarkers to detect animals with metabolic dysbalances such as SARA. MicroRNAs (miRNAs) are a class of small non-coding RNAs, serving as regulators of a plethora of molecular processes. To test our hypothesis, we performed a pilot study with non-lactating Holstein-Friesian cows fed a forage diet (FD; 0% concentrate, n = 4) or a high-grain diet (HG; 65% concentrate, n = 4) to induce SARA. Comprehensive profiling of miRNA expression in plasma and leucocytes were performed by next generation sequencing (NGS). The success of our model to induce SARA was evaluated based on ruminal pH and was evidenced by increased time spent with a pH threshold of 5.8 for an average period of 320 min/d. RESULTS: A total of 520 and 730 miRNAs were found in plasma and leucocytes, respectively. From these, 498 miRNAs were shared by both plasma and leucocytes, with 22 miRNAs expressed exclusively in plasma and 232 miRNAs expressed exclusively in leucocytes. Differential expression analysis revealed 10 miRNAs that were up-regulated and 2 that were down-regulated in plasma of cows when fed the HG diet. A total of 63 circulating miRNAs were detected exclusively in the plasma of cows with SARA, indicating that these animals exhibited a higher number and diversity of circulating miRNAs. Considering the total read counts of miRNAs expressed when fed the HG diet, differentially expressed miRNAs ( log2 fold change) and known function, we have identified bta-miR-11982, bta-miR-1388-5p, bta-miR-12034, bta-miR-2285u, and bta-miR-30b-3p as potential candidates for SARA-biomarker in cows by NGS. These were further subjected to validation using small RNA RT-qPCR, confirming the promising role of bta-miR-30b-3p and bta-miR-2285. CONCLUSION: Our data demonstrate that dietary change impacts the release and expression of miRNAs in systemic circulation, which may modulate post-transcriptional gene expression in cows undergoing SARA. Particularly, bta-miR-30b-3p and bta-miR-2285 might serve as promising candidate biomarker predictive for SARA and should be further validated in larger cohorts.

Short-chain fatty acids promote jejunal barrier function and caecal muscle contractibility in laying hens ex vivo.

1. Short-chain fatty acids (SCFA) exert beneficial actions in the gut; nevertheless, information about the effect of SCFA on physiological responses in the small intestine of chickens is rare.2. The aim of this study was to assess the effect of 1) different molar acetate:butyrate ratios (Ac:But; Experiment 1; 78.5% acetate and 7.3% butyrate versus 71.4% acetate and 14.0% butyrate) and 2) SCFA concentrations (Experiment 2; final concentration in chambers: 70.5 versus 141 µmol SCFA/ml buffer) on the jejunal and caecal contractibility and jejunal barrier function in laying hens. The change in muscle contractibility due to the SCFA was measured in mid-jejunal and caecal segments (n = 4 each per hen) from four laying hens using the organ bath system after precontraction with acetylcholine for 15 min. Changes in short-circuit current (ISC) and transepithelial tissue conductivity (GT) as indicators for net ion flux and barrier function, respectively, were measured in mid-jejunal tissue (n = 3/hen and treatment), mounted into Ussing chambers.3. In Experiment 1, the addition of SCFA, irrespective of the Ac:But ratio, decreased jejunal muscle tension (P < 0.05), jejunal GT as well as caused a less negative ISC (P < 0.05). In Experiment 2, the increasing SCFA concentrations increased the caecal muscle contraction and jejunal ISC by 75.6% while decreasing the GT by up to 19.6% (P < 0.05).4. In conclusion, results demonstrate that increasing butyrate proportions and SCFA concentrations stimulate caecal muscle contraction, thereby increasing caecal mixing and emptying in vivo. Jejunal ISC and GT support a strong SCFA sensing capacity in the jejunum, as both, more butyrate and higher SCFA, increased mucosal ion uptake and barrier function.

Cinnamon bark oil and coconut oil emulsions modified small intestinal motility and barrier function in laying hens in an ex vivo experiment.

1. Plant extracts and oils are supplemented in diets for chickens due to their antimicrobial capacities; however, little information exists whether they influence intestinal motility and barrier function.2. The present study aimed to determine the effect of increasing levels of cinnamon bark oil (CBO; 0%, 0.038%, 0.076% and 0.151%) and coconut oil emulsions prepared with soy and sunflower lecithin on the contractile function of enteric wall muscles in the jejunum and ileum and jejunal barrier function in laying hens.3. For testing muscle contraction, mid-jejunal and ileal segments (n = 4 each per hen) from four laying hens were placed in a longitudinal orientation into isolated organ baths filled with Krebs buffer and fastened to force transducers. Muscle segments were induced to contract with acetylcholine and the effects of the oil emulsions on contraction were measured.4. For barrier function, distal jejunal pieces were stripped of serosa before mounting into Ussing chambers and recording changes in short-circuit current (ISC) and transepithelial tissue conductivity (GT) before and after addition of the respective emulsion.5. The CBO decreased the muscle tone, representing a relaxation of on average 36.2% and 42.6% for the jejunum and ileum, respectively, compared to before the addition (P < 0.001). Moreover, CBO linearly decreased the ISC and GT of the jejunal mucosa, indicating a greater absorption of anions and increased barrier function (P < 0.001). Only the coconut oil-sunflower lecithin emulsion relaxed the muscles, whereas both coconut oil-lecithin emulsions increased the ISC but reduced the GT of the jejunal mucosa, which suggested an increased cation absorption and decreased paracellular permeability, respectively (P < 0.05).6. In conclusion, CBO and coconut oil-lecithin emulsions showed the potential to increase jejunal barrier function, whereas CBO may be more efficacious to slow down digesta passage in the small intestine.

Evaluating the structural effects of intra-articular sprifermin on cartilage and non-cartilaginous tissue alterations, based on sqMRI assessment over 2 years.

OBJECTIVE: Sprifermin (recombinant human fibroblast growth factor-18), a potential disease-modifying osteoarthritis (OA) drug, demonstrated dose-dependent effects on femorotibial cartilage thickness (by quantitative magnetic resonance imaging [MRI]) in the phase II FORWARD study. This post-hoc analysis evaluated the potential effects of sprifermin on several articular structures in the whole joint over 24 months using semi-quantitative MRI assessment. DESIGN: Patients aged 40-85 years with symptomatic radiographic knee OA, Kellgren-Lawrence grade 2 or 3, and medial minimum joint space width ≥2.5 mm in the target knee were randomized (1:1:1:1:1) to receive three double-blinded, once-weekly, intra-articular injections of sprifermin 30 µg or 100 µg or placebo every 6 (q6mo) or 12 months. 1.5- or 3 T MRIs were read using the Whole-Organ Magnetic Resonance Imaging Score (WORMS) system at baseline and 24 months. Change from baseline at 24 months on compartment and/or whole knee level was assessed for cartilage morphology, bone marrow lesions (BMLs), and osteophytes by delta-subregional and delta-sum (DSM) approaches. Menisci, Hoffa-synovitis, and effusion-synovitis were also evaluated for worsening. RESULTS: 549 patients were included. Dose-dependent treatment effects from baseline to 24 months were observed on cartilage morphology (sprifermin 100 µg q6mo vs placebo; mean DSM (95% confidence interval [CI]) -0.6 (-1.5, 0.2); less cartilage worsening) in the entire knee and BMLs sprifermin 100 µg q6mo vs placebo; mean DSM (95% CI) -0.2 (-0.5, 0.1) in the patellofemoral compartment. No effects over 24 months were observed on osteophytes, menisci, Hoffa-synovitis or effusion-synovitis. CONCLUSIONS: Positive effects associated with sprifermin were observed for cartilage morphology changes, and BML improvement. There were no meaningful negative or positive effects associated with sprifermin in the other joint tissues examined.

Predicting knee replacement in participants eligible for disease-modifying osteoarthritis drug treatment with structural endpoints.

OBJECTIVE: Evaluate associations between 2-year change in radiographic or quantitative magnetic resonance imaging (qMRI) structural measures, and knee replacement (KR), within a subsequent 7-year follow-up period. METHOD: Participants from the Osteoarthritis Initiative were selected based on potential eligibility criteria for a disease-modifying osteoarthritis (OA) drug trial: Kellgren-Lawrence grade 2 or 3; medial minimum joint space width (mJSW) ≥2.5 mm; knee pain at worst 4-9 in the past 30 days on an 11-point scale, or 0-3 if medication was taken for joint pain; and availability of structural measures over 2 years. Mean 2-year change in structural measures was estimated and compared with two-sample independent t-tests for KR and no KR. Area under the receiver operating characteristic curve (AUC) was estimated using 2-year change in structural measures for prediction of future KR outcomes. RESULTS: Among 627 participants, 107 knees underwent KR during a median follow-up of 6.7 years after the 2-year imaging period. Knees that received KR during follow-up had a greater mean loss of cartilage thickness in the total femorotibial joint and medial femorotibial compartment on qMRI, as well as decline in medial fixed joint space width on radiographs, compared with knees that did not receive KR. These imaging measures had similar, although modest discrimination for future KR (AUC 0.62, 0.60, and 0.61, respectively). CONCLUSIONS: 2-year changes in qMRI femorotibial cartilage thickness and radiographic JSW measures had similar ability to discriminate future KR in participants with knee OA, suggesting that these measures are comparable biomarkers/surrogate endpoints of structural progression.

Relationship of Plasminogen Activator Inhibitor 1 4G/5G Gene Polymorphism and Nontraumatic Lunatum Avascular Necrosis.

PURPOSE: Plasminogen activator inhibitor 1 (PAI-1) is a critical enzyme that regulates coagulation and fibrinolytic systems. The aim of this study was to determine the role of PAI-1 4G/5G polymorphism in nontraumatic avascular necrosis of the lunate. METHODS: The study included 45 patients with Kienböck disease and 45 healthy individuals as a control group. In both groups, genomic DNA was extracted from peripheral blood samples to determine the distributions of PAI-1 4G/5G polymorphism using allele-specific polymerase chain reaction and sequencing. RESULTS: No statistically significant difference was determined in the distribution of the gene polymorphism between the patient and control groups. We found the 5G/5G genotype to be 1.7 times higher in the control group compared with the patient group. A 1.6-fold increase in the 4G homozygote genotype was identified in the patient group. The patient and control groups were also evaluated for 4G/4G plus 4G/5G and 5G/5G in terms of genotype distribution. No statistically significant difference was found. CONCLUSIONS: The findings suggest that the PAI-1 4G/4G polymorphism is not a genetic risk for Kienböck disease. CLINICAL RELEVANCE: This study aimed to reveal the genetic etiology of Kienböck disease.

SP1-mediated downregulation of ADAMTS3 gene expression in osteosarcoma models.

ADAMTS3 is a member of procollagen N-proteinase subfamily of ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) gene family. It has an important function in the procollagen maturation process. The removal of N-peptidases is required for the accurate processing of fibrillar collagens. Otherwise, several disorders can occur that is related with the collagenous tissues. ADAMTS3 mainly maturates type II collagen molecule which is the main component of the bone and cartilage. There are several expression studies about ADAMTS3 gene however its transcriptional regulation has not been lightened up, yet. Here we first time cloned and functionally analyzed the promoter region of ADAMTS3 gene, approximately 1380 bp upstream of the transcription start site. Transient transfection experiments showed that all truncated promoter constructs are active and 171 bp fragment is sufficient to activate gene expression in both Saos-2 and MG63 cells. In silico analysis showed that ADAMTS3 has a TATA-less promoter and contains several SP1/GC box binding motifs and a CpG island. Therefore we mainly investigated the SP1 dependent regulation of ADAMTS3 promoter. SP1 downregulated ADAMTS3 transcriptional activity. As consistent with the transcriptional activity, mRNA, and protein expression levels were also decreased by SP1. On the other hand, functional binding of the SP1 on multiple regions of ADAMTS3 promoter was confirmed by EMSA studies. As ADAMTS3 is responsible for the collagen maturation and biosynthesis, further we investigated the effect of SP1 on type I-II and III collagen gene expressions. We point out that SP1 increased type II and III collagen expression and in contrast decreased type I collagen expression levels in Saos-2 cells. mRNA expression level was decreased for all collagen types in MG63 model. Decrease in the type II collagen expression was also demonstrated at the protein level by SP1. Collectively these results provide first findings for the SP1-related transcriptional regulation of ADAMTS3 and collagen genes in osteosarcoma cell lines.

Induction of human ADAMTS-2 gene expression by IL-1α is mediated by a multiple crosstalk of MEK/JNK and PI3K pathways in osteoblast like cells.

Up-regulation of ADAMTS genes with proinflammatory cytokines is important for some pathological conditions such as osteoarthritis (OA) that is a disease based on ECM degradation in cartilage. IL-1α is a proinflammatory cytokine and important both to normal and pathophysiologic conditions in cartilage and bone. Effects of some proinflammatory cytokines such as TNF-α and IL-1ß on the some members of ADAMTS family have been investigated in some chondrocyte tissues or cell lines. However the effect of the IL-1α on the expression of ADAMTS-2 and ADAMTS-3 gene expression in osteoblast like cell lines, remains unclear. Therefore, the aim of this study is to investigate the effect of IL-1α on ADAMTS-2 and ADAMTS-3 gene expression in osteoblast like cells, Saos-2 and MG-63. The present study, for the first time, demonstrated that IL-1α increases ADAMTS-2 and ADAMTS-3 gene expressions in both Saos-2 and MG-63 cells. Having correlation to mRNA induction, the upregulation of ADAMTS-2,-3 protein levels by IL-1α stimulation is also observed. The inhibition studies showed that this upregulation occurred at the level of transcription, and there was no effect of IL-1α on ADAMTS-2 mRNA half-life in Saos-2 cells. Transactivation potential of IL-1α on ADAMTS-2 promoter was investigated by transient transfection assay. Specifically, IL-1α strongly increased -658/+112 and -530/+112 ADAMTS-2 promoter constructs. Further, we analyzed signaling pathways involved in ADAMTS-2 induction. Pathway inhibition studies revealed that this upregulation depends on the activation of MEK, JNK and PI3K pathways. These findings suggested that IL-1α is a strong positive regulator of ADAMTS-2 and ADAMTS-3 expression. These findings would provide novel insight into the pathophysiology of OA.

Physiologic load-bearing characteristics of autografts, allografts, and polymer-based scaffolds in a critical sized segmental defect of long bone: an experimental study.

BACKGROUND: To address the challenge of treating critical sized intercalary defects, we hypothesized that under physiologic cyclic loading, autografts, allografts, and scaffolds loaded with and without human mesenchymal stem cells (hMSCs) would have different biomechanical characteristics. METHODS: Using a rat femoral defect model, 46 rats were assigned to four groups, ie, autograft (n = 12), allograft (n = 10), scaffold (n = 13), and scaffold with hMSCs (n = 11). The scaffold groups used a 5 mm segment of scaffold composed of 80% poly-ε-caprolactone and 20% hydroxyapatite. Rats were sacrificed 4 months postoperatively, and the repairs were assessed radiographically and biomechanically. RESULTS: Autograft and allograft groups exhibited the most bridging callus, while the scaffold/hMSCs group had more callus than the scaffold repairs. Although signs of radiographic healing did not accurately reflect restoration of mechanical properties, addition of hMSCs on the scaffold enhanced bone formation. The scaffold alone group had significantly lower elastic and viscous stiffness and higher phase angles than other repairs and the contralateral controls. Addition of hMSCs increased the elastic and viscous stiffness of the repair, while decreasing the phase angle. CONCLUSION: Further comparative analysis is needed to optimize clinical use of scaffolds and hMSCs for critical sized defect repairs. However, our results suggest that addition of hMSCs to scaffolds enhances mechanical simulation of native host bone.

Shear flows at the tokamak edge and their role in core rotation and the L-H transition.

Pfirsch-Schlüter fluxes in tokamaks are shown to drive strong poloidal and toroidal shear flows that are localized to the edge and scrape-off layer in the presence of temperature gradients and finite bootstrap current in the pedestal. Within a magnetohydrodynamic model, the effect of these flows on core rotation and their role in the magnetic configuration dependence of the power threshold for the low- (L-) to high- (H-)mode transition are discussed. Theoretical predictions based on symmetries of the underlying equations, coupled with computational results, are found to be in general agreement with observations in the Alcator C-Mod tokamak [Phys. Plasmas 12, 056111 (2005)10.1063/1.1876294].