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High-density assembly of gold nanoparticles on multiwalled carbon nanotubes using 1-pyrenemethylamine as interlinker.

In this article, we describe the formation of carbon nanotube (CNT)-gold nanoparticle composites in aqueous solution using 1-pyrenemethylamine (Py-CH2NH2) as the interlinker. The alkylamine substituent of 1-pyrenemethylamine binds to a gold nanoparticle, while the pyrene chromophore is noncovalently attached to the sidewall of a carbon nanotube via pi-pi stacking interaction. Using this strategy, gold nanoparticles with diameters of 2-4 nm can be densely assembled on the sidewalls of multiwalled carbon nanotubes. The formation of functionalized gold nanoparticles and CNT-Au nanoparticle composites was followed by UV-vis absorption and luminescence spectroscopy. After functionalization of gold nanoparticles with 1-pyrenemethylamine, the distinct absorption vibronic structure of the pyrene chromophore was greatly perturbed and its absorbance value was decreased. There was also a corresponding red shift of the surface plasmon resonance (SPR) absorption band of the gold nanoparticles after surface modification from 508 to 556 nm due to interparticle plasmon coupling. Further reduction of the pyrene chromophore absorbance was observed upon formation of the CNT-Au nanoparticle composites. The photoluminescence of 1-pyrenemethylamine was largely quenched after attaching to gold nanoparticles; formation of the CNT-Au nanoparticle composites further lowered its emission intensity. The pyrene fluoroprobe also sensed a relatively nonpolar environment after its attachment to the nanotube surface. The present approach to forming high-density deposition of gold nanoparticles on the surface of multiwalled carbon nanotubes can be extended to other molecules with similar structures such as N-(1-naphthyl)ethylenediamine and phenethylamine, demonstrating the generality of this strategy for making CNT-Au nanostructure composites.

Journal Article↗

Quantitation of metal content in the silver-assisted growth of gold nanorods.

The seed-mediated approach to making gold nanorods in aqueous surfactant solutions has become tremendously popular in recent years. Unlike the use of strong chemical reductants to make spherical gold nanoparticles, the growth of gold nanorods requires weak reducing conditions, leading to an unknown degree of gold reduction. The metal content of gold nanorods, made in high yield in the presence of silver ion, is determined by inductively coupled plasma atomic emission spectroscopy. Through the use of the known gold concentration in nanorods, molar extinction coefficients are calculated for nanorods of varying aspect ratios from 2.0 to 4.5. The extinction coefficients at the longitudinal plasmon band peak maxima for these nanorods vary from 2.5x10(9) to 5.5x10(9) M-1 cm-1, respectively, on a per-particle basis. Many of the gold ions present in the growth solution remain unreacted; insights into the growth mechanism of gold nanorods are discussed.

Journal Article↗

Spectroscopic study of laser-induced phase transition of gold nanoparticles on nanosecond time scales and longer.

The pulsed laser induced phase transition of gold nanoparticles in aqueous solution was observed via a transient absorption on nanosecond time scales and longer. Gold nanoparticles were excited with an intense picosecond laser pulse (355 nm, 30 ps), and the subsequent changes were monitored using two continuous wave laser wavelengths (488 and 635 nm). On the nanosecond time scale, below 6.3 mJ cm(-2), no change was observed; however, in the low fluence region between 6.3 and 17 mJ cm(-2), gold nanoparticles produced a bleach signal (488 nm) attributed to the melting of the gold nanoparticles, which decreased linearly with increasing laser fluence. Laser fluences above 17 mJ cm(-2) resulted in a strong absorption at both wavelengths, which is ascribed to vaporization of gold nanoparticles rather than solvated electrons (ejected from gold nanoparticles) or light scattering. The decay of both signals was faster than the 5 ns time resolution used in our experimental system. On the microsecond time scale, increase in absorbance at 635 nm was observed with a time constant of 1.0 micros, while no change was observed at 488 nm. It is considered that this increase is attributed to the formation of smaller gold nanoparticles resulting from pulsed laser induced size reduction of initial gold nanoparticles.

Journal Article↗

Assessment of 4-(dimethylamino)pyridine as a capping agent for gold nanoparticles.

Gold nanoparticles stabilized with 4-(dimethylamino)pyridine (DMAP) were prepared by ligand exchange and phase transfer (toluene/water) of functionalized gold nanoparticles. DMAP-protected gold nanoparticles are water-soluble, positively charged, and fairly monodisperse (6.2 +/- 0.9 nm). To understand the scope of this interesting system, the details of the binding of DMAP to gold nanoparticles were investigated. The adsorption of DMAP onto gold surfaces was studied by electrochemistry and surface plasmon resonance. It is concluded that of the three most likely binding modes, the one involving the pyridine nitrogen binding to the gold surface, as suggested previously (Gittins, D. I.; Caruso, F. Angew. Chem., Int. Ed. 2001, 40, 3001), is consistent with experimental data. Other 4-substituted pyridines were also assessed as capping agents. The solubility in toluene and basicity of the incoming ligand, as well as the ability to form charged nanoparticles, determine whether ligand exchange and subsequent phase transfer of the nanoparticles occur. The solubility and stability of the DMAP-protected gold nanoparticles were studied as a function of pH using UV-visible spectroscopy and transmission electron microscopy (TEM). These nanoparticles are soluble and stable over a wide pH range (5.0-12.8). It was found that excess DMAP is necessary for both the preparation and the stability of the DMAP-protected gold nanoparticles.

Journal Article↗

Thermoresponsive behavior of poly(n-isopropylacrylamide) hydrogels containing gold nanostructures.

We report the changes in the structure and thermoresponsive behavior of poly(N-isopropylacrylamide) (PNIPAm) hydrogels when gold nanostructures are synthesized in situ within the hydrogel matrix. Cross-linked PNIPAm hydrogels were synthesized using NIPAm and 0.00-3.50% (w/w versus NIPAm) of N,N'-methylenebisacrylamide (MBAm) and/or N,N'-cystaminebisacrylamide (CBAm) as cross-linking agents. The hydrogels were soaked in potassium tetrachloroaurate to introduce gold ions. The hydrogels containing Au3+ were then immersed in a sodium borohydride solution to reduce the gold ions. Infrared spectroscopy, UV-visible spectroscopy, and equilibrium swelling were used to examine the structural/physical differences between gels of different compositions; UV-visible spectroscopy and mass measurements were used to observe the kinetics and thermodynamics of the hydrogel volume phase transition. These studies revealed several differences in the physical characteristics and thermoresponsive behavior of hydrogels based on cross-linker identity and the presence or absence of gold nanostructures. Hydrogels with gold nanostructures and high CBAm and low MBAm content have equilibrium swelling masses 3-20 times their native analogues. In comparison, gold-containing hydrogels with high MBAm and low CBAm content have swelling masses that are equal to their native analogues. Additionally, the gold-containing PNIPAm hydrogels cross-linked with only CBAm have a deswelling temperature of approximately 40 degrees C, approximately 8 degrees C above the samples cross-linked with only MBAm. Varying the CBAm content and introducing gold enables tuning of the deswelling temperature.

Journal Article↗

Generation of gold thread from Au(III) and triethylamine.

Gold threads were spontaneously generated at room temperature from a combined solution of aqueous HAuCl4 and triethylamine. Initially, some reduced gold nanoparticles were self-organized into one-dimensional (1D) nanowires, which then developed into three-dimensional (3D) microscale threads of gold. After aging, the self-organized gold thread grew spontaneously, accompanied by the generation of new buds to start new branches. TEM images show that the gold thread is composed of elastic materials; in which, colloidal gold nanoparticles reduced by triethylamine serve as the bricks, whereas elastic substances formed during the reaction serve as the mortar, which holds together the gold particles. The results of IR and 1H NMR spectroscopy demonstrate that the elastic materials consist mainly of alkane-like substances. Examination of the 1H NMR spectrum of triethylamine under the conditions of the reaction shows evidence of the occurrence of oxidative N dealkylation. The observed isotope effect demonstrates the existence of O-D cleavage resulting from the degradation of carbinolamine, which is the reported oxidation route of amines. Triethylamine acts here not only acts as a stabilizer and a reducing agent, but also as a precursor to build the alkane-like material upon which the gold thread is formed.

Journal Article↗

Intracellular colocalization of variant surface glycoprotein and transferrin-gold in Trypanosoma brucei.

Endocytosis and intracellular transport has been studied in the bloodstream forms of Trypanosoma brucei by light and electron microscopy, using colloidal gold coupled to bovine transferrin (transferrin-gold). The endocytosed transferrin-gold, visualized by silver intensification for light microscopy, was present in vesicular structures between the cell nucleus and flagellar pocket of the organism. At the ultrastructural level, transferrin-gold was present after a 10-min incubation in the flagellar pocket, coated vesicles, cisternal networks, and lysosomelike structures. Endocytosis and intracellular processing of T. brucei variable surface glycoprotein (VSG) was studied using two preparations of affinity-purified rabbit IgG directed against different parts of the VSG. One preparation of IgG was directed against the cross-reacting determinant (CRD): a complex glycolipid side chain covalently linked to the COOH-terminus of the VSG molecule. The other was directed against determinants on the rest of the VSG molecule. When the two IgG preparations were used on thawed, thin cryosections of trypanosomes that had been incubated in transferrin-gold before fixation, the organelles involved with transferrin-gold endocytosis labeled with both antibodies, as well as many vesicular, tubular, and vacuolar structures that did not contain endocytosed transferrin-gold. Both antibodies also labeled the cell surface. In double-labeling experiments both antibodies were closely associated except that IgG directed against the VSG molecule labeled all the cisternae of the Golgi apparatus, whereas anti-CRD IgG was shown to label only half of the Golgi apparatus. Evidence for sorting of VSG molecules from endocytosed transferrin-gold was found. Double-labeling experiments also showed some tubular profiles which labeled on one side with anti-CRD IgG and on the other side with anti-VSG IgG, suggesting a possible segregation of parts of the VSG molecule.

Animals↗

Long-term results of MISTI gold breast implants: a retrospective study.

MISTI Gold breast implants (Bioplasty, St. Paul, Minn.) filled with polyvinylpyrrolidone-hydrogel were developed as a promising alternative to silicone-filled implants. Some studies have reported on the positive effects of the implant, such as improved radiolucency and biocompatibility of the gel; however, there are also reports that such implants increased in volume and were subject to capsular contracture in the human body, resulting in demands for their removal. The purpose of this retrospective study was to analyze the long-term results of a series of patients with MISTI Gold breast implants. Between 1991 and 1993, the authors inserted 83 MISTI Gold implants in 61 patients with an average age of 46 years (range, 16 to 69). The authors were able to follow up 48 patients with 71 MISTI Gold implants. The average follow-up was 68 months (range, 10 to 108 months). The retrospective study found that 59 percent of all MISTI Gold implants were removed after an average period of 4.14 years. The main reason for implant removal was an increase in volume of 38 percent, followed by capsular contracture in 14 percent of all 71 MISTI Gold implants. The average increase in volume of all removed MISTI Gold implants was 43 percent. Capsular contracture was graded as Baker I and II in 63 percent and as Baker III and IV in 37 percent. In conclusion, the authors believe that MISTI Gold implants do not fulfill the criteria of safe breast implants, and they agree with the December of 2000 opinion of the Medical Devices Agency of the Department of Health in London that the hydrogel-filled breast implants should not be used until more information about the filler material and its metabolic fate is available.

Adolescent↗

X-ray absorption spectroscopic studies on gold nanoparticles in mesoporous and microporous materials.

Au L(3)-edge X-ray absorption spectroscopic measurements were carried out over a series of mesoporous and microporous materials containing gold nanoparticles to investigate the effects of the host matrix and preparation methods on the properties of gold nanoparticles. The materials of structure type MCM-41, ZSM-5, SAPO-18 and LSX with varying framework composition containing low concentrations of gold nanoparticles were prepared and characterized. In these materials the size of the gold nanoparticles varied in the range approximately 1 to 4 nm. A series of gold nanoparticles within different mesoporous and microporous materials have been investigated using X-ray absorption fine structure (XANES, EXAFS) and other techniques. Information such as atomic distances, bonding and neighbouring environment obtained from XAFS measurements was useful in elucidating the nature and structure of gold nanoparticles on these catalytic materials. The influence of the high-temperature (823, 1113, 1273 K) treatment on gold nanoparticles inside the mesoporous matrix was investigated using the XAFS technique. The XAFS and XANES results confirm various characteristics of gold nanoparticles in these materials suitable for catalysis, fabrication of nanodevices and other applications.

Journal Article↗

Colloidal Gold Cytochemistry of Endo-1,4-beta-Glucanase, 1,4-beta-D-Glucan Cellobiohydrolase, and Endo-1,4-beta-Xylanase: Ultrastructure of Sound and Decayed Birch Wood.

Colloidal gold coupled to endo-1,4-beta-glucanase II (EG II) and 1,4-beta-D-glucan cellobiohydrolase I (CBH I), isolated from Trichoderma reesei (QM9414), and endo-1,4-beta-xylanase from Aureobasium pullulans (NRRLY-2311-1) was used successfully to determine the ultrastructural localization of cellulose and xylan in sound birch wood. In addition, these enzyme-gold complexes demonstrated the distribution of cellulose and xylan after decay by three white rot fungi, Phanerochaete chrysosporium, Phellinus pini, and Trametes versicolor, and one brown rot fungus, Fomitopis pinicola. Transverse sections of sound wood showed that EG II was localized primarily on the S(1) layer of the secondary wall, whereas CBH I labeled all layers of the secondary wall. Oblique sections showed a high concentration of gold labeling, using EG II or CBH I. Preference for the sides of the microfibrillar structure was observed for both EG II and CBH I, whereas only CBH I had a specificity for the cut ends of microfibrils. Labeling with the xylanase-gold complex occurred primarily in the inner regions of the S(2) layer, S(1), and the middle lamella. In contrast, little labeling occurred in the middle lamella with EG II or CBH I. Intercellular regions within the cell corners of the middle lamella were less electron dense and labeled positively when EG II- and xylanase-gold were used. Wood decayed by P. chrysosporium or P. pini was delignified, and extensive degradation of the middle lamella was evident. The remaining secondary walls labeled with EG II and CBH I, but little labeling was found with the xylanase-gold complex. Wood decayed by T. versicolor was nonselective, and erosion of all cell wall layers was apparent. Remaining wall layers near sites of erosion labeled with both EG II and CBH I. Erosion troughs that reached the S(1) layer or the middle lamella had less xylanase-gold labeling in the adjacent cell wall that remained. Brown-rotted wood had very low levels of gold particles present in sections treated with EG II or xylanase. Labeling with CBH I had the lowest concentrations in the S(2) layer near cell lumina and corresponded to sites with the most extensive degradation.

Journal Article↗

Decreased level of antibodies against Helicobacter pylori in patients with rheumatoid arthritis receiving intramuscular gold.

BACKGROUND: Sodium aurothiomalate has been reported to have in vitro activity against Helicobacter pylori. Intramuscular gold, as given to patients with rheumatoid arthritis (RA), may therefore influence the colonisation of the gastric mucosa with H pylori. METHODS: Two groups were compared. One group of 42 patients was treated with intramuscular gold; the other group of 58 patients was treated with antimalarial drugs. Antibodies to H pylori (IgA and IgG) were assessed by an enzyme linked immunosorbent assay (ELISA) and total IgA and IgG were measured by nephelometry. RESULTS: IgA and IgG antibody titres against H pylori and total IgA and IgG levels were lower in the patients treated with gold than in the group treated with antimalarial drugs. The ratio of IgA antibodies to H pylori to total IgA antibodies and the ratio of IgG antibodies to H pylori to total IgG antibodies were lower in the group treated with gold. The percentage of seropositivity to H pylori was significantly lower in the group treated with gold than in the group treated with antimalarial drugs for the two IgA antibodies (35 and 55% respectively) and IgG antibodies to H pylori (40 and 65% respectively). CONCLUSIONS: Although this study cannot completely exclude the possibility that a suppressive effect of intramuscular gold on total immunoglobulin production plays a part in the decrease in the titres of IgA antibodies to H pylori and IgG antibodies to H pylori, the lower ratios of antibodies to H pylori to total immunoglobulin antibodies and the lower percentages of seropositivity to H pylori in the group treated with gold suggests that treatment with intramuscular gold decreases H pylori colonisation.

Adolescent↗

Insulin response to an intravenous glucose load during development of obesity in gold thioglucose-injected mice.

The insulin secretory response to an intravenous glucose load was examined in chronically catheterized, conscious mice 2, 5, and 10 wk after induction of obesity by a single injection of gold thioglucose. At 2 wk after administration of gold thioglucose, a significant increase in both the insulinemia and incremental area under the curve of insulin release after intravenous glucose were observed (incremental area under the curve for 2-wk control mice, 852 +/- 54 min/pM; incremental area under the curve for 2-wk GTG-injected mice, 1140 +/- 114 min/pM; P < 0.05). At this stage, no significant difference existed in the glucose tolerance or body weight of control and gold thioglucose-injected mice. By 5 wk, the gold thioglucose-injected mice were approximately 33% heavier than their lean controls and showed a marked glucose intolerance. This was accompanied by overt hyperinsulinemia in both the basal state and also in response to an intravenous glucose bolus as indicated by the increase in the incremental area under the curve of insulin (5-wk control mice, 816 +/- 114 min/pM; 5-wk gold thioglucose-injected mice, 1374 +/- 156 min/pM; P < 0.05). At 10 wk after gold thioglucose administration, body weight and the degree of glucose intolerance were increased. Although 10-wk gold thioglucose-injected mice showed basal hyperinsulinemia, an intravenous glucose bolus elicited a smaller insulin secretory response than that observed in the age-matched lean control animals (10-wk control mice, 672 +/- 54 min/pM; 10-wk gold-thioglucose-injected mice 186 +/- 42 min/pM; P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗

[Possible role of cellular immunity against tubular basement membrane antigen (TBM) in gold nephropathy in rheumatoid arthritis patients].

Autoantibody formation and lymphocytes proliferative response to tubular basement membrane (TBM) antigen were examined to clarify the pathogenesis of gold nephropathy, in rheumatoid arthritis patients. The existence of tubulopathy was ascertained by urine protein analysis, electrophoresis and urine TBM antigen titration. Circulating antibody to human TBM antigen titrated by enzyme immunoassay was significantly elevated in patients with gold tubulopathy, and mitogenic stimulation with TBM antigen of peripheral lymphocytes specifically responded in the early stage after receiving gold, but then clearly decreased after the cessation of gold. But, when the lymphocytes had been passed through a nylon wool column, the reaction was remarkably high even in the later stage after receiving gold, suggesting that another suppressive population of lymphocytes became trapped in the nylon wool column. This evidence suggests that gold compounds definitely act as initiating and promoting agents, and the development of tubular disorders induced by gold are likely related to the cellular recognition of effector T cells to the TBM antigen, following the strong effect of gold on the cellular immune system.

Adult↗

Longterm outcome of treatment of Felty's syndrome with intramuscular gold: case reports and recommendations for management.

OBJECTIVE: To evaluate the incidence, complications, and course of Felty's syndrome (FS) in patients treated with intramuscular (IM) gold. METHODS: Retrospective chart review of all FS cases (1979 to 2003) was conducted in the Mary Pack Arthritis Centre (MPAC) gold clinic. FS was diagnosed if patients had rheumatoid arthritis (RA; American College of Rheumatology criteria) and persistent leukopenia [white blood cell (WBC) count < 4] in the absence of other known causes of leukopenia. Splenomegaly was not part of the inclusion criteria. RESULTS: Thirteen patients with FS were identified in the gold clinic population. The mean age at diagnosis of FS was 58.7 years and the mean duration of RA at time of diagnosis was 6.9 years. The weekly dose of gold ranged from 10 mg to 50 mg depending on tolerability. Gold therapy resulted in normalization of the WBC count in 9 of 13 patients. The mean time to normalization of the WBC was 40 weeks. Only one patient with FS had experienced recurrent infectious complications from FS, and this did not recur after gold treatment was initiated. No patient had vasculitis. CONCLUSION: In our gold clinic population FS is a mild disease and is rarely associated with infectious complications. Gold is an effective treatment of FS.

Adult↗

Gold is a T cell polyclonal activator in BN and LEW rats but favors IL-4 expression only in autoimmune prone BN rats.

Gold salts are beneficial in the treatment of rheumatoid arthritis but may induce immune-mediated disorders in predisposed patients. Gold salts induce Th2-dependent autoimmunity in Brown-Norway (BN) rats but not in Lewis (LEW) rats. The aim of this study was to define molecular targets of gold salts and to approach why LEW rats are resistant. Gold salts act on early steps of transduction in T cells from BN and LEW rats since they trigger tyrosine phosphorylation of numerous proteins including p56(lck) and a calcium signal which results in IL-4 and IFN-gamma expression by BN and LEW T cells. However, the IL-4 response was favored in BN spleen cells in vitro and in vivo. IFN-gamma, produced in part by CD8(+) cells, contributes to the resistance of LEW rats since gold salt-injected LEW rats receiving anti-CD8 or anti-IFN-gamma mAb displayed the parameters characteristics of gold salt-induced Th2 autoimmunity although to a lesser extent than in BN rats. Gold salts transduce a signal in BN and LEW spleen cells resulting in IL-4 and IFN-gamma gene transcription with a preferential IL-4 response in BN rats, a Th2-prone strain, while IFN-gamma contributes to the resistance of LEW rats.

Animals↗

Oral gold: a comparison with placebo and with intramuscular sodium aurothiomalate.

The therapeutic and toxic effects of the orally absorbed gold compound auranofin have been compared with placebo and parenterally administered sodium aurothiomalate (GST) in 90 patients with active rheumatoid arthritis over one year. At the end of this period 57% of patients on auranofin, 73% on GST and 0% on placebo remained on therapy. Toxicity caused discontinuation of therapy in 10% of patients on auranofin, 20% on GST and 7% on placebo. Lack of response to therapy led to withdrawal of 20% on auranofin, 0% on GST and 90% on placebo. Multiple clinical and biochemical assessments were performed during this study. Analysing them separately, and in the case of 6 of them grouped together in a disease activity index, the same trend is apparent throughout, namely that placebo has no effect on active inflammatory rheumatoid arthritis, and that both gold drugs are beneficial. GST has an earlier effect and tends to produce a greater change but after one year there was no significant difference between the 2 gold drugs for any parameter assessed. Gold levels in plasma or erythrocytes did not predict or correlate with either the development of toxicity or clinical efficacy. This study has demonstrated the second-line potential of auranofin which seems to be effective at gold concentrations in the blood below those observed with GST therapy. Toxicity limits the use of gold salts in RA. If the reduced incidence of adverse reaction with auranofin observed in this study is substantiated in larger numbers over prolonged periods, the use of gold at an earlier stage of disease may be facilitated.

Administration, Oral↗

Comparison of auranofin, gold sodium thiomalate, and placebo in the treatment of rheumatoid arthritis. Subsets of responses.

A comparison of placebo, auranofin, and parenteral gold sodium thiomalate therapy in 209 patients with active rheumatoid arthritis was performed in a 21-week prospective, controlled, double-blind multicenter trial. When the 161 patients who completed at least 20 weeks of treatment were analyzed for different degrees of response, no remissions were identified. When 50 percent or greater improvement of pain/tenderness scores were compared for end of trial versus entry values, 9 percent of placebo-treated patients, 34 percent of auranofin-treated patients, and 48 percent of gold sodium thiomalate-treated patients showed important improvement that was statistically significant for both gold treatments. When 50 percent improvement for joint swelling was analyzed, 12 percent of the placebo-treated group, 28 percent in the auranofin-treated group, and 37 percent in the gold sodium thiomalate-treated group showed this degree of improvement. Auranofin almost achieved statistical significance for improvement in joint swelling when compared with placebo (p = 0.07), but gold sodium thiomalate was much better than placebo (p = 0.009). There was no statistically significant difference between the two gold treatments. Thus it appears that a subset of patients had an important response to gold therapy that would not be evident by the usual analyses of mean or median changes. Analysis for predictors of response did not discriminate between responders and nonresponders. Because the trial was limited to 21 weeks of therapy, no prediction of the longer-term-effects, especially for auranofin, should be inferred.

Adolescent↗

Gold concentrations in blood fractions of patients with rheumatoid arthritis treated with Myocrisin.

Gold levels in the plasma and blood cells of patients treated with the gold drug Myocrisin (sodium aurothiomalate) were determined by atomic absorption spectrometry. There is a correlation between whole blood gold and plasma gold concentrations which is different for smokers and non-smokers. Most cellular gold is associated with the membrane and is present in concentrations approximately equivalent to the number of reactive sulphydryl groups on the exofacial surface of the cell. Since gold would be expected to react with SH groups and since these groups are vital for cellular function, a possible role for gold in modifying cellular metabolism is indicated.

Arthritis, Rheumatoid↗