Evaluation of the Impact of a New Synthetic Vitamin E-Bonded Membrane on the Hypo-Responsiveness to the Erythropoietin Therapy in Hemodialysis Patients: A Multicenter Study.

BACKGROUND: Oxidative stress has been related to hypo-response to erythropoiesis-stimulating agents (ESAs) in hemodialysis (HD) patients. The aim of this study was to verify whether vitamin E (ViE) on a synthetic polysulfone dialyzer can improve ESA responsiveness. METHODS: This controlled, multicenter study involved 93 HD patients on stable ESA therapy, who were randomized to either ViE-coated polysulfone dialyzer or to a low-flux synthetic dialyzer. The primary outcome measure was the change in ESA resistance index (ERI) from baseline. RESULTS: Mean ERI decreased in the ViE group by 1.45 IU/kg*g/dl and increased in the control group by 0.53 IU/kg*g/dl, with a mean difference of 1.98 IU/kg*g/dl (p = 0.001 after adjusting for baseline ERI, as foreseen by the study protocol). Baseline ERI was inversely related to its changes during follow-up only in the control group (R2 = 0.29). CONCLUSIONS: The ViE dialyzer can improve ESA response in HD patients. Changes in ERI during follow-up are independent from baseline ERI only in the ViE group. Video Journal Club 'Cappuccino with Claudio Ronco' at http://www.karger.com/?doi=453442.

High-volume online haemodiafiltration improves erythropoiesis-stimulating agent (ESA) resistance in comparison with low-flux bicarbonate dialysis: results of the REDERT study.

BACKGROUND: In haemodialysis (HD) patients, anaemia is associated with reduced survival. Despite treatment with erythropoiesis-stimulating agents (ESAs), a large number of patients with chronic kidney disease show resistance to this therapy and require much higher than usual doses of ESAs in order to maintain the recommended haemoglobin (Hb) target, and recent studies suggest that hepcidin (HEP) may mediate the ESA resistance index (ERI). High-volume online haemodiafiltration (HV-OL-HDF) has been shown to improve anaemia and to reduce the need for ESAs in HD patients; this effect is associated with a reduced inflammatory state in these patients. The aim of the REDERT study (role of haemodiafiltration on ERI) was to investigate the effect of different dialysis techniques on ERI and HEP levels in chronic dialysis patients. METHODS: A single cross-over, randomized, multicentre study (A-B or B-A) was designed. Forty stable HD patients from seven different dialysis units (male 65%, mean age 67.6 ± 14.7 years and mean dialytic age 48 ± 10 months) were enrolled. Patients were randomized to the standard bicarbonate dialysis (BHD) with low-flux polysulfone (PS) membrane group or to the HV-OL-HDF group with high-flux PS membranes and exchange volume of >20 L/session. After 6 months, patients were shifted to the other dialytic group for a further 6 months. Clinical data, Hb, ESA doses and iron metabolism were recorded every month. HEP, beta2-microglobulin (b2MG) and C-reactive protein (CRP) were determined every 3 months, and ERI was calculated monthly as the weekly ESA dose per kilogram of body weight divided by Hb level. Data were analysed using paired-samples t-test, Wilcoxon signed-rank test and Spearman's correlation coefficient. RESULTS: Dialysis efficiency for small molecules assessed as Kt/V was significantly increased in HV-OL-HDF from 1.47 ± 0.24 to 1.49 ± 0.16; P < 0.01. A significant reduction of b2MG was obtained in HV-OL-HDF from month 3 whereas CRP values were not significantly changed during the study period either in BHD or HV-OL-HDF.ERI was significantly reduced in HV-OL-HDF at month 3 and 6 (from 9.1 ± 6.4 UI/weekly/Kg/Hb to 6.7 ± 5.3 UI/weekly/Kg/Hb; P < 0.05) due to a higher ESA consumption in BHD in spite of similar Hb levels. HEP levels were reduced in HV-OL-HDF with respect to BHD after 3 and 6 months. Iron consumption was not significantly different during BHD or HV-OL-HDF treatment as well as transferrin, ferritin and TSAT levels. A significant positive linear correlation between HEP and ERI (r(2) = 0.258, P < 0.001) was observed. CONCLUSIONS: In a uraemic patient population with low-grade inflammation treated with HV-OL-HDF, we observed a significant reduction of ERI values as well as HEP levels. The positive correlation between these two parameters supports a role for HEP in the development of ERI in the dialytic population. Moreover, the lower b2MG and the higher Kt/V achieved in HV-OL-HDF confirms the better depurative effect of this technique in comparison with BHD with respect to middle molecules and small-molecular-weight molecules.

Distribution of Elements in Durum Wheat Seed and Milling Products: Discrimination between Cultivation Methods through Multivariate Data Analysis.

Many staple foods originate from durum wheat and its milling products; because of this, it is very important to know their characteristics. This study investigates elemental contents in these products and if differences exist because of organic farming. The concentrations of 28 elements in the whole seed and in milling products, that is, bran, semolina and flour, of durum wheat, were determined through ICP-OES. The wheats were grown under conventional or organic agronomic practices to verify the possibility of discriminating, using the elemental content, between products coming from one or the other practice. The elements were more abundant in the outer layer of the seed, the bran, but most of them were also present in the others. Traces of Sb were present only in 3% of the samples, while traces of Tl were detected in approximately half of the seed and bran samples but not in other samples. The absence of an element was more characteristic of specific products, e.g., most semolina and flour lacked Co, while other elements showed small differences between products from organic and conventional cultivation or between different milling products, which was the case, for example, for traces of Ag, B, and V. The concentrations of these elements were coupled with multivariate discriminant analysis, specifically PLS-DA, to identify the cultivation provenance of the milled products. A few elements, although different for each product, are sufficient to attain precision and accuracy of classification close to 1; small differences exist for different products. The worst is flour, where the predicted precision and accuracy are 0.92, although using only three elements: B, K, and Se. Semolina attains perfect prediction when also adding to the three previous elements, Ag, Cd, and Cu. Further elements are necessary for bran, while Fe and Mg replace K and Ag to classify seeds. In conclusion, five elements, B, Cd, Cu, K, and Se, are the most important in distinguishing between organic and conventional agriculture; these elements also permit some differentiation among products. The method could help in fraud prevention.

ON-SITE monitoring OF BVOCS emission in Tremiti island, Italy.

A measurement campaign was conducted on San Domino Island, part of the Tremiti Islands archipelago, located in Foggia, Italy. The area is almost entirely covered by vegetation, dominated by the following main species: Juniperus turbinata, Helichrysum italicum, Myrtus communis, Rosmarinus officinalis, Pistacia lentiscus and Pinus halepensis.This study focused on the BVOCs emitted by plants and the ground, employing a simple, economical, and efficient sampling and analysis method. The main known BVOC species emitted by Mediterranean plant species as α-pinene, ß-pinene, camphene and limonene were detected. The measurements highlighted a daily complementarity between plant and soil emissions. The daily variations in BVOCs emitted by both plants and the soil are differ, ensuring an almost constant concentration throughout the day. At the same time, the composition of sea spray aerosol (SSA) was also measured. The measurement sites were selected based on botanical characterization to account for the predominant species on San Domino Island, and the sampling was conducted at human height to accurately identify the species for potential use. The combination of beneficial effects of the substances emitted by plant species and soil, along with the simultaneous presence of SSA, are factors that could enhance the effectiveness of forest therapy in a previously unexplored location.

Nutritional knowledge in hemodialysis patients and nurses: focus on phosphorus.

OBJECTIVE: To assess the knowledge of adult hemodialysis patients and nurses working in dialysis units, specifically with regard to knowledge of phosphorus and other nutrients related to dietary management of end-stage renal disease. DESIGN: Cross-sectional cohort study. SETTING: Hemodialysis unit. SUBJECTS: One hundred ninety-one hemodialysis patients and 105 dialysis nurses, as well as 86 control hospital employees who are not health professionals. INTERVENTION: Nutritional knowledge was assessed by a 25-item chronic kidney disease knowledge assessment tool for nutrition, which includes 15 questions on phosphorus and 10 questions on protein, sodium, and potassium knowledge. RESULTS: The scores obtained by patients were much lower than those of nurses (11.6 ± 3.9 vs. 16.0 ± 2.2, P < .001) but slightly higher than those of controls (10.6 ± 3.2, P < .05). Patients with phosphorus serum level >5.5 mg/dL showed chronic kidney disease knowledge assessment tool for nutrition scores similar to those of patients with a serum phosphorus level <5.5 mg/dL. The prevalence of right answers to questions regarding knowledge of phosphorus was lower than that regarding knowledge of the other nutrients, both for patients (38.4% ± 17.8% vs. 57.3% ± 19.9%, P < .001) and nurses (55.6% ± 11.1% vs. 74.8% ± 11.7%, P < .001) as well as for controls (30.7% ± 14.5% vs. 60.1% ± 17.4%, P < .001). CONCLUSIONS: Our study suggests that nutritional knowledge of hemodialysis patients, although higher than the general population, is lower for phosphorus with respect to the other nutrients, such as protein, sodium, and potassium. This occurs even in patients with hyperphosphatemia or those taking phosphate binder medications. Nurses showed the best scores; however, improvement is necessary, especially with regard to knowledge of phosphorus. Training programs on nutrition for nurses and on information for patients should be implemented. They can contribute to achievement of a more effective control of phosphate balance, reduction of costs, and improvement of the quality of care for hemodialysis patients.

[Paired hemodiafiltration]. / Paired Hemodiafiltration (PHF).

Paired hemodiafiltration (PHF) is a dialysis modality where an ultrafilter and a filter with hollow-fiber membranes are used in series for on-line infusion of ultrapure fluid. PHF is a on-line hemodiafiltration modality which allows reinfusion using the dialysate prepared by the monitor. The dialysate is infused by retrofiltration, which takes place in the filter positioned in series with regard to the filter where ultrafiltration and the diffusive/convective exchange of solutes occur. The indications and contraindications of the method are the same as those of standard on-line hemodiafiltration, but PHF minimizes the inflammatory response of the patient. This is the result of the increased number of ultrafiltration stages (three), two of which in the monitor and the third by the filter in series, where the retrofiltration takes place. The method can also be carried out with acetate-free dialysate; this reduces the side effects of acetate used to stabilize pH in bicarbonate dialysate, resulting in better cardiovascular stability during dialysis.

Disclosing the Redox Pathway Behind the Excellent Performance of CuS in Solid-State Batteries.

Copper sulfide has attracted increasing attention as conversion-type cathode material for, especially, solid-state lithium-based batteries. However, the reaction mechanism behind its extraordinary electroactivity is not well understood, and the various explanations given by the scientific community are diverging. Herein, the CuS reaction dynamics are highlighted by examining the occurring redox processes via a cutting-edge methodology combining X-ray absorption fine structure spectroscopy, and chemometrics to overcome X-ray diffraction limitations posed by the poor material's crystallinity. The mathematical approach rules out the formation of intermediates and clarifies the direct conversion of CuS to Cu in a two-electron process during discharge and reversible oxidation upon delithiation. Two distinct voltage regions are identified corresponding to Cu- as well as the S-redox mechanisms occurring in the material.

Efficacy, Safety and Pharmacoeconomic Analysis of Intravenous Ferric Carboxymaltose in Anemic Hemodialysis Patients Unresponsive to Ferric Gluconate Treatment: A Multicenter Retrospective Study.

Patients undergoing hemodialysis with iron deficiency anemia (IDA) receiving treatment with erythropoiesis-stimulating agents (ESAs) who were intolerant or non-responsive to intravenous (i.v.) ferric gluconate (FG) (hemoglobin; Hb values < 10.5 g/dL or increase in <1 g/dL) or % transferrin saturation; TSAT of <20%) in the previous 6 months were switched to i.v. ferric carboxymaltose (FCM). Changes in iron status parameters, economic and safety measures were also assessed. Seventy-seven hemodialysis patients aged 68 ± 15 years were included. Erythropoietin resistance index decreased from 24.2 ± 14.6 at pre-switch to 20.4 ± 14.6 after 6 months of FCM treatment and Hb levels ≥10.5 g/dL improved from 61% to 75.3% patients (p = 0.042). A 1 g/dL increase in Hb levels was also seen in 26% of patients as well as a 37.7% increase in patients achieving >20% increase in TSAT after FCM. Levels of Hb, TSAT and ferritin parameters increased during FCM treatment with a concomitant decrease in ESA. A mixed-model analysis, which also considered gender, confirmed these trends. Safety variables remained stable, no hypersensitivity reaction was recorded and only one patient reported an adverse event after FCM. FCM treatment was associated with a cost saving of 11.11 EUR/patient/month. These results confirm the efficacy, safety and cost-effectiveness of FCM in correcting IDA in hemodialysis patients.

Multivariate curve resolution analysis for interpretation of dynamic Cu K-edge X-ray absorption spectroscopy spectra for a Cu doped V(2)O(5) lithium battery.

Vanadium pentoxide materials prepared through sol-gel processes act as excellent intercalation hosts for lithium as well as polyvalent cations. A chemometric approach has been applied to study the X-ray absorption near-edge structure (XANES) evolution during in situ scanning of the Cu(0.1)V(2)O(5) xerogel/Li ions battery. Among the more common techniques, the fixed size windows evolving factor analysis (FSWEFA) permits the number of species involved in the experiment to be determined and the range of existence of each of them. This result, combined with the constraints of the invariance of the total concentration and non-negativity of both concentrations and spectra, enabled us to obtain the spectra of the pure components using a multivariate curve resolution refined by an alternate least squares fitting procedure. This allowed the normalized concentration profile to be understood. This data treatment evidenced the occurrence, for the first time, of three species during the battery charging. This fact finds confirmation by comparison of the pure spectra with the experimental ones. Extended X-ray absorption fine structure (EXAFS) analysis confirms the occurrence of three different chemical environments of Cu during battery charging.

Economic impact of ferric carboxymaltose in haemodialysis patients / Impatto economico dell'utilizzo di carbossimaltosio ferrico nei pazienti in emodialisi

Intravenous iron supplementation is essential in hemodialysis (HD) patients to recover blood loss and to meet the requirements for erythropoiesis and, in patients receiving erythropoietin, to avert the development of iron deficiency. In a recent real-world study, Hofman et al. showed that a therapeutic shift from iron sucrose (IS) to ferric carboxymaltose (FCM) in HD patients improves iron parameters while reducing use of iron and erythropoietin. The objective of this economic analysis is to compare the weekly cost of treatment of FCM vs IS in hemodialysis patients in Italy. The consumption of drugs (iron and erythropoietin) was derived from Hofman's data, while the value was calculated at Italian ex-factory prices. The analysis was carried on the total patient sample and in two subgroups: patients with iron deficiency and patients anemic at baseline. In addition, specific sensitivity analyses considered prices currently applied at the regional level, simulating the use of IS vs iron gluconate (FG) and epoetin beta vs epoetin alfa. In the base-case analysis, the switch to FCM generates savings of -€12.47 per patient/week (-21%) in all patients, and even greater savings in the subgroups with iron deficiency -€17.28 (-27%) and in anemic patients -€23.08 (-32%). Sensitivity analyses were always favorable to FCM and confirmed the robustness of the analysis. FCM may represent a cost-saving option for the NHS, and Italian real-world studies are needed to quantify the real consumption of resources in dialysis patients.

Effect of Water and Alkali-Ion Content on the Structure of Manganese(II) Hexacyanoferrate(II) by a Joint Operando X-ray Absorption Spectroscopy and Chemometric Approach.

Manganese hexacyanoferrate (MnHCF) is made of earth-abundant elements by a safe and easy synthesis. The material features a higher specific capacity at a higher potential than other Prussian blue analogs. However, the effect of hydration is critical to determine the electrochemical performance as both the electrochemical behavior and the reaction dynamics are affected by interstitial/structural water and adsorbed water. In this study, the electrochemical activity of MnHCF is investigated by varying the interstitial ion content through a joint operando X-ray absorption spectroscopy and chemometric approach, with the intent to assess the structural and electronic modifications that occur during Na release and Li insertion, as well as the overall dynamic evolution of the system. In MnHCF, both the Fe and Mn centers are electrochemically active and undergo reversible oxidation during the interstitial ion extraction (Fe2+ /Fe3+ and Mn2+ /Mn3+ ). The adsorption of water results in irreversible capacity during charge but only on the Fe site, which is suggested by our chemometric analysis. The local environment of Mn experiences a substantial yet reversible Jahn-Teller effect upon interstitial ion removal because of the formation of trivalent Mn, which is associated with a decrease of the equatorial Mn-N bond lengths by 10 %.

Chemical and Sensory Profiling of Monovarietal Extra Virgin Olive Oils from the Italian Marche Region.

Chemical and sensory peculiarities of monovarietal extra virgin olive oils (MEVOOs) from the cultivars (cvs.) Ascolana tenera (ASC), Coroncina (COR), Mignola (MIG), Piantone di Mogliano (MOG), and Raggia (RAG) from Marche region (Italy) are investigated. Their polar phenolic substances and α-tocopherol are analysed through high performance liquid chromatography with different detectors. Volatile substances, fatty acid composition, and squalene are analysed by gas chromatography coupled to mass spectrometry (MS) and to the flame ionization detector, respectively. Total antioxidant activity and sensory analysis were also performed. MOG showed high squalene content (on average 0.88 ± 0.16 g/100 g), high relative amount of α-copaene among volatiles, and the highest oleic acid percentage. MIG had high α-tocopherol content (on average 350.0 ± 57.6 mg kg-1) and high α-farnesene in the volatile fraction. ASC showed the highest sensory quality and the lignan pinoresinol with higher concentration as compared to the other MEVOOs (p < 0.05), which resulted in a possible chemical marker for this cv. RAG was characterized by the sensory note of almond, which corresponds to its highest (E)-2-hexenal percentage. Sensory analysis and an antioxidant activity assay performed on a set of industrial extra virgin olive oils purchased in supermarkets, highlighted MEVOOs' superiority from these points of view. Principal component analysis displays the main characteristics of the cvs. investigated.

Nonpeptidergic allosteric antagonists differentially bind to the CXCR2 chemokine receptor.

The chemokine receptor CXCR2 is involved in different inflammatory diseases, like chronic obstructive pulmonary disease, psoriasis, rheumatoid arthritis, and ulcerative colitis; therefore, it is considered an attractive drug target. Different classes of small CXCR2 antagonists have been developed. In this study, we selected seven CXCR2 antagonists from the diarylurea, imidazolylpyrimide, and thiazolopyrimidine class and studied their mechanisms of action at human CXCR2. All compounds are able to displace (125)I-CXCL8 and inhibit CXCL8-induced beta-arrestin2 recruitment. Detailed studies with representatives of each class showed that these compounds displace and antagonize CXCL8, most probably via a noncompetitive, allosteric mechanism. In addition, we radiolabeled the high-affinity CXCR2 antagonist SB265610 [1-(2-bromophenyl)-3-(4-cyano-1H-benzo[d] [1,2,3]-triazol-7-yl)urea] and subjected [(3)H]SB265610 to a detailed analysis. The binding of this radioligand was saturable and reversible. Using [(3)H]SB265610, we found that compounds of the different chemical classes bind to distinct binding sites. Hence, the use of a radiolabeled low-molecular weight CXCR2 antagonist serves as a tool to investigate the different binding sites of CXCR2 antagonists in more detail.

Multibody Models for the Analysis of a Fall From Height: Accident, Suicide, or Murder?

The final subject position is often the only evidence in the case of the fall of a human being from a given height. Foreseeing the body trajectory and the respective driving force may not be trivial due to the possibility of rotations and to an unknown initial position and momentum of the subject. This article illustrates how multibody models can be used for this aim, with specific reference to an actual case, where a worker fell into a stair well, prior to stair mounting, and he was found in an unexpected posture. The aim of the analysis was establishing if this worker was dead in that same place, if he had been pushed, and which was his initial position. A multibody model of the subject has been built ("numerical android"), given his stature and his known mass. Multiple simulations have been performed, following a design of experiments where various initial positions and velocity as well as pushing forces have been considered, while the objective function to be minimized was the deviation of the numerical android position from the actual worker position. At the end of the analysis, it was possible to point how a very limited set of conditions, all including the application of an external pushing force (or initial speed), could produce the given final posture with an error on the distance function equal to 0.39 m. The full analysis gives a demonstration of the potentiality of multibody models as a tool for the analysis of falls in forensic inquiries.

Direct quantification of test bacteria in synthetic water-polluted samples by square wave voltammetry and chemometric methods.

A home-made microelectrode array, based on reticulated vitreous carbon, was used as working electrode in square wave voltammetry experiments to quantify the bacterial load of Escherichia coli ATCC 13706 and Pseudomonas aeruginosa ATCC 27853, chosen as test microorganisms, in synthetic samples similar to drinking water (phosphate buffer). Raw electrochemical signals were analysed with partial least squares regression coupled to variable selection in order to correlate these values with the bacterial load estimated by aerobic plate counting. The results demonstrated the ability of the method to detect even low loads of microorganisms in synthetic water samples. In particular, the model detects the bacterial load in the range 3-2,020 CFU ml(-1) for E. coli and in the range 76-155,556 CFU ml(-1) for P. aeruginosa.

Chemical and sensory differences between high price and low price extra virgin olive oils.

The aim of the study was to identify new potential chemical markers of extra virgin olive oil (EVOO) quality by using a multicomponent analysis approach. Sixty-six EVOOs were purchased from the Italian market and classified according to their price as low price EVOOs (LEVOOs) and high price EVOOs (HEVOOs) costing 3.60-5.90euro/L and 7.49-29.80euro/L respectively. Sensory and chemical parameters strictly related to olive oil quality have been investigated, like volatile substances, polar phenolic substances, antioxidant activity, fatty acid composition, and α-tocopherol. Significant differences in terms of chemical composition and sensory features have been highlighted between the two EVOOs classes investigated, proving a generally lower level of quality of LEVOOs, clearly showed also by means of principal component analysis. Among the most interesting outcomes, R ratio (free tyrosol and hydroxytyrosol over total free and bound forms), measuring the extent of secoiridoids hydrolysis, resulted to be significantly higher in LEVOOs than in HEVOOs. Other key differences were found in the volatile substances composition, in the stearic acid percentage and in p-coumaric acid content.

Correction: Interaction of healthcare staff's attitude with barriers to physical activity in hemodialysis patients: A quantitative assessment.

[This corrects the article DOI: 10.1371/journal.pone.0196313.].

Interaction of healthcare staff's attitude with barriers to physical activity in hemodialysis patients: A quantitative assessment.

BACKGROUND AND AIM OF THE STUDY: In hemodialysis patients, sedentarism is a potentially modifiable mortality risk factor. We explored whether healthcare staff's attitude towards exercise interacts with patient-perceived barriers in modifying the level of physical activity in this population. METHODS: In this prospective, cross-sectional, multicenter study we recruited 608 adult patients and 330 members of the healthcare staff in 16 hemodialysis units in Italy. We assessed patient-perceived barriers to, and healthcare staff's attitude towards, exercise by specific questionnaires. We fitted multilevel linear models to analyze the relationships of either barriers or staff's attitude, and their interaction, with a measure of patient self-reported physical activity (the Human Activity Profile-Adjusted Activity Score [HAP-AAS]), adjusting for multiple confounders. We also employed latent class analysis to dichotomize patients into those endorsing or not endorsing barriers. RESULTS: Most barriers were negatively associated with the HAP-AAS (adjusted change attributable to a given barrier ranging between -5.1 ["Feeling too old", 95% Confidence Interval: -9.4 to -0.8] and -15.6 ["Ulcers on legs and feet", 95%CI: -24.8 to -6.5]. We found a significant interaction between staff's attitude and barriers (adjusted P values ranging between 0.03 ["I do not believe that it is physician's or nurse's role providing advice on exercise to patients on dialysis"] and 0.001 ["I do not often ask patients about exercise"]). A beneficial effect of a proactive staff's attitude was evident only in patients not endorsing barriers. CONCLUSIONS: Barriers and non-proactive staff's attitude reduce physical activity in hemodialysis patients. Patients not endorsing barriers benefit the most from a proactive staff's attitude.

Deconvolution of UV-VIS spectroelectrochemical thin-layer data for an EE electrochemical process.

A deconvolution method, based on Fixed Size Window Evolving Factor Analysis (FSWEFA) has been developed to analyse the spectroelectrochemical sequence of spectra obtained in OTTLE electrodes for EE electrochemical processes. The analysis of the data permits to reconstruct the concentration profiles, the spectra of the intermediates and an optical analogue of the i vs E curve.

Complete inhibition of rhTSH-, Graves' disease IgG-, and M22-induced cAMP production in differentiated orbital fibroblasts by a low-molecular-weight TSHR antagonist.

The TSH receptor (TSHR) on orbital fibroblasts (OF) is a proposed target of the autoimmune attack in Graves' ophthalmopathy. In the present study, we tested whether the novel low-molecular-weight (LMW) TSHR antagonist Org-274179-0 inhibits cAMP production induced by rhTSH, Graves' disease IgG (GD-IgG), or M22 (a potent human monoclonal TSHR stimulating antibody) in cultured and differentiated OF from Graves' ophthalmopathy patients. cAMP production significantly increased after incubation either with 10 mU/ml rhTSH (3-fold; P ≤ 0.05), 1 mg/ml GD-IgG (2-fold; P ≤ 0.05), or 500 ng/ml M22 (5-fold; P ≤ 0.05). Incubation with the LMW TSHR antagonist dose dependently inhibited rhTSH, GD-IgG as well as the M22-induced cAMP production at nanomolar concentrations; complete blockade was affected at 10(-6) M. Our results suggest that GD-IgG- and M22-induced cAMP production in differentiated OF is exclusively mediated via the TSHR because it can be completely blocked by the LMW TSHR antagonist, Org 274179-0.