Comparison of Lutetium-177 tin colloid and Rhenium-188 tin colloid radiosynovectomy in chronic knee arthritis.

OBJECTIVE: To assess the role of Lutetium-177(Lu-177) tin colloid for radiosynovectomy and compare it with Rhenium-188 (Re-188) tin colloid radiosynovectomy for alleviation of pain in patients with chronic inflammatory arthritis of knee. METHODS: Patients of chronic inflammatory arthritis of the knee underwent pretherapeutic evaluation in a form of knee ultrasonogram, bone scan and clinical evaluation. Fifty-seven recruited patients were allocated at random to receive either intraarticular injections of Lu-177 tin colloid or Re-188 tin colloid. Eventually, 27 patients received Re-188 tin colloid and 30 patients received Lu-177 tin colloid. The joint was then immobilized for 2 days. Response evaluation was done using knee ultrasound, bone scan and clinical findings. RESULT: Of 30, 20 patients responded to radiosynovectomy in the Lu-177 tin colloid group compared to 21/27 patients in the Re-188 tin colloid group. CONCLUSION: Lu-177 tin colloid is an effective alternative to Re-188 tin colloid for radiosynovectomy in patients with chronic inflammatory knee arthritis.

Cu2ZnSnS4 nanocrystals for microwave thermal and microwave dynamic combination tumor therapy.

Cu2ZnSnS4 nanocrystals (CZTS NCs) have been demonstrated to be effective in tumor therapy as a novel susceptible agent for microwave thermal and microwave dynamic therapy. CZTS NCs intensify the heating effect of microwaves with a significant temperature increase of about 15 °C compared to the control group and showed remarkable anti-tumor performance after 5 min of microwave irradiation. For the first time, we report the microwave absorption performance and singlet oxygen production of CZTS NCs used in microwave therapy, which reveals new opportunities for novel combined mechanisms of microwave thermal and microwave dynamic tumor therapies.

The efficacy of tin-filtration for computed tomography in diagnosing urolithiasis.

INTRODUCTION: The purpose of this study was to evaluate the radiation dose and image quality of computed tomography urograms (CTU) using tin-filtration compared to conventional CTU (without tin-filtration) examinations in patients with suspected urolithiasis. METHODS: Group 1 consisted of 100 patients who were examined using the tin-filtered CTU protocols (Sn100kVp or Sn150kVp); Group 2 consisted of 100 patients who were examined using the same protocols but without tin-filtration (GE-NI41 or GE-NI43). The scanning protocol was based on the patients' body weight (<80 kg and ≥80 kg). The effective doses of all scans were compared between the two groups. Subjective image quality was evaluated by two blinded radiologists. The objective image quality was assessed for noise, signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR) and figure-of-merit (FOM) using the CTU scans acquired from both the tin-filtered and non-tin-filtered protocols. RESULTS: Tin-filtration resulted in the reduction of effective radiation dose ranging between 72% to 88% for the ≥80 kg and <80 kg patient groups respectively. For both groups, tin-filtration resulted in no significant differences in SNR and a significant increase in FOM. For the <80 kg group, tin-filtration resulted in significantly noisier images but with no significant difference in CNR. For the ≥80 kg group, tin-filtration resulted in significantly higher CNR. There was no significant difference in subjective image quality when assessed by the radiologists in terms of diagnostic confidence for urolithiasis. CONCLUSION: Tin-filtration significantly reduces patient dose while maintaining diagnostic image quality of CTUs for patients with suspected urolithiasis.

Recent Advances in Remineralization Therapies for Caries Lesions.

Remineralization of caries lesions is naturally achieved by salivary ions, and it can be enhanced by external factors or elements such as fluoride. Numerous studies have demonstrated the remineralizing efficacy of fluoride therapies as well as the limitations with some groups of the population. Consequently, developing new remineralization therapies to close this gap in efficacy has been a priority for the last 2 decades. In this review, we summarize and briefly discuss some of the latest advances in remineralization therapies. Most new therapies try to enhance the effect of fluoride by adding other potentially active ingredients to the formulation, such as calcium, phosphate, stannous, xylitol, and arginine. Other remineralization strategies have focused on creating remineralizing scaffolds within the lesions (e.g., self-assembling peptides). While several of the new remineralization strategies have progressed significantly in recent years, for most of them, the evidence is still insufficient to assess their true clinical potential.

A comprehensive biological insight of trinuclear copper(II)-tin(IV) chemotherapeutic anticancer drug entity: in vitro cytotoxicity and in vivo systemic toxicity studies.

Cisplatin (cis-diamminedichloroplatinum(II), CDDP) causes severe systemic toxicity, which limits its application in cancer treatment. Nevertheless, incorporation of endogenously present essential metal ions (copper) in anticancer drug regimes in a heterometallic ligand scaffold can substantially modulate the toxic effects of non-essential metals (platinum), thereby reducing unwanted toxic side effects. A chiral l-tryptophan derived [bis(1,2-diaminobenzene) copper(II)] chloride complex [CuSn2(Trp)] was previously synthesized by us as an active chemotherapeutic agent. Furthermore, we have explored CuSn2(Trp) induced in vitro cytotoxicity in a panel of human cancer cell lines and in vivo acute and systemic toxicities in healthy female Rattus norvegicus (Wistar) rats. MTT assay showed that CuSn2(Trp) exhibits strong anticancer potency against ovarian (PA-1) and prostate carcinomas (PC-3) but lower potency towards liver (HepG2) and breast carcinomas (MCF-7). Further, flow cytometric analysis demonstrated that CuSn2(Trp) kills PA-1 cells dose-dependently after 48 h treatment. Fluorescence microscopy and western blotting revealed that the plausible mechanism behind CuSn2(Trp) cytotoxicity was apoptosis, which was substantiated by cleavage of caspase-3 and poly-(ADP-ribose) polymerase (PARP). Furthermore, it has lower toxicity than CDDP in rats as evident from its eight fold (98.11 mg kg(-1)) more medial lethal dose (LD50) than CDDP (12 mg kg(-1)). Besides, the safety profile of CuSn2(Trp) was also established and no measurable DNA damage, nephrotoxicity, hepatotoxicity and neurotoxicity were observed when assessed as a function of oxidative stress markers in contrast to CDDP at equivalent lower doses. Our findings are of high importance in the context of further in vivo cancer studies on the CuSn2(Trp) drug entity.

Role of rhenium-188 tin colloid radiosynovectomy in patients with inflammatory knee joint conditions refractory to conventional therapy.

AIM: To evaluate the treatment response of rhenium-188 (¹88Re) tin colloid radiosynovectomy (¹88Re-RSV) in patients with inflammatory knee joint conditions refractory to conventional treatment. MATERIALS AND METHODS: Sixty-one knee joints in 48 patients with chronic synovitis caused by various inflammatory knee joint diseases refractory to conventional therapy were included in this prospective study. All patients were assessed clinically for pain, tenderness, joint swelling, mobility, analgesic intake, and blood pool activity on bone scan. Different scores were assigned to all these parameters. RSV of knee joint was done using intra-articular injection of 555 MBq of ¹88Re tin colloid. Response was assessed at 3, 6, and 12 months using various clinical parameter scores and blood pool bone scan mentioned above and categorized as responders and nonresponders on the basis of change in percentage of cumulative scores. RESULTS: Of the 61 joints, 57 were responder and four were nonresponder at 3-month post-RSV. Out of 57 responders, seven had recurrence on further follow-up (three at 6 months and four at 12 months). There was a statistically significant reduction in clinical parameters cumulative scores at 3, 6, and 12 months when compared with baseline (P<0.0001) in responder group. Blood pool scinitgraphy also showed decrease in blood pool activity compared with the baseline. There was statistically significant association between the responder group and shorter duration of disease (P=0.011). Patients having normal or minor X-ray changes, little or no swelling, mild tenderness, and better mobility were independently associated with good response. CONCLUSION: ¹88Re tin colloid synovectomy is a useful treatment modality in patients with chronic inflammatory knee joint conditions refractory to conventional treatment. Patients with shorter duration of disease, normal or minor X-ray findings, little or no swelling, mild tenderness, and better mobility are better candidates for RSV.

Tin and fluoride as anti-erosive agents in enamel and dentine in vitro.

OBJECTIVE: To investigate the efficacy of an experimental tin- and fluoride-containing mouth rinse on progression of erosion in enamel and dentine in vitro. MATERIAL AND METHODS: Human enamel and dentine specimens were subjected to a cyclic demineralization and remineralization procedure for 10 days, with six 5-min demineralization periods per day. Erosive demineralization was performed with 0.05 M citric acid (pH 2.3). Except in the negative control group, the specimens were treated for 2 min with mouth rinses after the first and sixth demineralizations. An experimental tin-containing fluoride mouth rinse [125 mg/kg F(-) (amine fluoride), 375 mg/kg F(-) (NaF), 800 mg/kg Sn(2+) (SnCl(2))] and an experimental sodium fluoride mouth rinse (500 mg/kg F(-)) were used (both pH 4.5). A commercially available, tin-containing mouth rinse served as a positive control (pH 4.2, 409 mg/kg Sn(2+), 250 mg/kg F(-)). Tissue loss was determined profilometrically. RESULTS: The highest tissue loss was found in the negative control group, in both enamel and dentine. In enamel, the NaF solution showed almost no effect. Both tin-containing solutions significantly reduced tissue loss (positive control: 65%; 800 mg/kg Sn(2+): 78%; both p ≤ 0.001 compared to negative control). In dentine all mouth rinses significantly reduced tissue loss (positive control: 43%; 800 mg/kg Sn(2+): 53%; NaF: 40%; all p ≤ 0.001 compared to negative control). CONCLUSIONS: In enamel, the efficacy of mouth rinses depended on the compound used; tin-containing preparations were notably effective. In dentine, however, reduction of substance loss was nearly the same in all treatment groups.

Characterization of Re-188-Sn microparticles used for synovitis treatment.

Rhenium-188 labeled tin (Sn) microparticles were developed for pain palliation therapy in the patients suffering from synovitis with acute pain. The rhenium tin microparticles were prepared using stannous chloride and freshly eluted (188)ReO(4)(-) from (188)W/(188)Re generator. The aggregated colloidal particles, packed in a spherical form after boiling for 90-120min were analyzed using electron microscope. The size, surface morphology and stability of microparticles were analyzed by changing temperature and volume conditions. The small colloidal particles clustered and formed spherical microparticles. The 90% of microparticles were in 5-10microm range, after 90min and 120min of boiling. The radiolabeling efficiency was improved to 98% after centrifugation for 10min at 3500rpm. The formulations were stable but the increase in volume had inverse effect on labeling efficiency. No leak was observed from knee area up to 24h with 15-20mCi injection of (188)Re-Sn microparticles. The relief in treated patients, from the pain and inflammation, was observed clinically and by (99m)Tc-MDP perfusion scan.

Tin foil as bolus material for therapeutic electron beams from the Varian Clinac 2100C/D.

Tin foils of sub-millimetre thickness have been investigated as bolus material for therapeutic electron beams from the Varian Clinac 2100C/D linear accelerator. Measurements with ionisation chamber and radiographic film in Plastic Water or water were performed under tin foil bolus to determine surface dose, therapeutic ranges, output factor correction, penumbra and dose outside the field edge. Appropriate thicknesses of tin foil for 90% dose at the surface were found to be approximately 0.3 mm for 6 MeV, and 0.4 mm for 9 MeV and 12 MeV. Enhanced therapeutic interval with tin foil bolus over water-equivalent bolus has previously been reported, but was found not to be evident for 12 MeV and for a small (4 x 4 cm2) 9 MeV field. The penumbra width of fields with tin foil and water-equivalent bolus were found to be within 2 mm, while the doses at 1 cm outside the field edge were within 1.5% of peak dose. Output factor corrections for fields with tin foil were measured as within 2% of unity. Air gaps between the tin foil and phantom surface up to 5 mm were observed to have minimal effect on output correction factor, relative surface dose, and therapeutic range.

188Re-tin-colloid as a new therapeutic agent for rheumatoid arthritis.

Radiation synovectomy is a useful treatment modality in patients with refractory synovitis. We have developed a 188Re-tin-colloid as a new radiopharmaceutical agent and investigated its efficacy and safety in patients with rheumatoid arthritis. Radiation synovectomy was performed using 188Re-tin-colloid in 22 knees from 21 rheumatoid arthritis patients refractory to intra-articular corticosteroid injection. The efficacy and safety of administration of 370-1110 MBq of 188Re-tin-colloid were evaluated after 1, 3, 6, 9 and 12 months. Pain intensity on a visual analogue scale decreased significantly 12 months after therapy (mean+/-SD: 68.0+/-26.1 mm vs. 25.1+/-23.4 mm; P=0.0001 by the paired t-test). Pain decreased in 19 cases (86.3%), joint tenderness improved in 14 cases (63.6%) and joint swelling was reduced in all cases (100%). 188Re-tin-colloid was safe. The residual activity of 188Re in the blood was 0.077%+/-0.25% of the injected dose. The radioactivity of 188Re in the urine was 0.14%+/-0.13% of the injected dose. Transient reactive synovitis was observed in 18 cases (81.8%). No clinical side-effects or abnormalities in leucocyte count, platelet count, liver function tests or urine analysis were observed in any patient. In conclusion, in this first study of radiation synovectomy using 188Re-tin-colloid for patients with rheumatoid arthritis, the treatment resulted in the improvement of arthritis and was well tolerated.

Radionuclide therapy with bone-seeking compounds: Monte Carlo calculations of dose-volume histograms for bone marrow in trabecular bone.

The purpose of the present work was to investigate how haematopoietic stem cell survival is affected by the differences in the dose distribution that arise from different radionuclides contained in bone-seeking radiopharmaceuticals. This was carried out in three steps: (a) calculations of representative dose distributions in individual bone marrow cavities that are irradiated by sources of 89Sr, 186Re, 117mSn or 153Sm, uniformly distributed on the bone surfaces; (b) assessment of the corresponding haematopoietic stem cell survival and (c) a comparison of these results with results obtained using the assumption of a uniform dose distribution. Two different idealized models of the geometry of trabecular bone were formulated, each consisting of an infinite array of identical elements. Monte Carlo simulations were used to generate dose-volume histograms that were used to assess haematopoietic stem cell survival with two different assumptions about spatial cell distributions. Compared with a homogeneous dose distribution, the estimated cell survival was markedly higher for 117mSn and 153Sm, and only slightly different for 89Sr and 186Re. The quantitative results differed between the two geometric models and the assumptions about spatial cell distribution, but the trends were the same. The results imply that it is necessary to include dose distributions for individual bone marrow cavities in considerations concerning bone marrow toxicity.

Considerations in the selection of radiopharmaceuticals for palliation of bone pain from metastatic osseous lesions.

UNLABELLED: Bone pain is a common complication for terminal patients with bone metastases from prostate, lung, breast, and other malignancies. A multidisciplinary approach in treating bone pain is generally required, 1 which includes a combination of analgesic drug therapy, radiation therapy, hormonal therapy, and chemotherapy. Over the years, treatment of bone pain using bone-seeking radiopharmaceuticals has been explored extensively. Pharmaceuticals labeled with energetic 1-particle emitters such as 32p, 89Sr, 153Sm, and 186Re, in addition to the low-energy electron emitter 117mSn, have been studied for this purpose. Bone-marrow toxicity as a consequence of chronic irradiation by the energetic , particles is a general problem associated with this form of treatment. It is therefore desirable to identify radiochemicals that minimize the dose to the bone marrow and at the same time deliver therapeutic doses to the bone. METHODS: New S values (mean absorbed dose per unit cumulated activity) for target regions of human bone and marrow were used to ascertain the capacity of various radiochemicals to deliver a high bone dose while minimizing the marrow dose. The relative dosimetric advantage of a given radiopharmaceutical compared with a reference radiochemical was quantitated as a dosimetric relative advantage factor (RAF). Several radionuclides that emit energetic 1 particles (32p, 89Sr, 153Sm, 186Re, and 177Lu) and radionuclides that emit low-energy electrons or beta particles (169Er, 117mSn, and 33p) were evaluated. For these calculations, ratios of the cumulated activity in the bone relative to cumulated activity in the marrow alpha equal to 10 and 100 were used. RESULTS: When the radiopharmaceutical was assumed to be uniformly distributed in the endosteum and alpha was taken as 100 for both the reference and test radiochemicals, the RAF values compared with the reference radionuclide 32p were 1.0, 1.2, 1.4, 1.6, 1.7, 1.9, and 2.0 for 89Sr, 186Re, 153Sm, 177Lu, 169Er, 117mSn, and 33P, respectively. In contrast, when the radiopharmaceutical is assumed to be uniformly distributed in the bone volume, the RAF values for these 7 radionuclides were 1.1, 1.5, 2.4, 3.2, 4.5, 5.1, and 6.5, respectively. CONCLUSION: These results suggest that low-energy electron emitters such as 117mSn and 33P are more likely to deliver a therapeutic dose to the bone while sparing the bone marrow than are energetic beta emitters such as 32p and 89Sr. Therefore, radiochemicals tagged with low-energy electron or beta emitters are the radiopharmaceuticals of choice for treatment of painful metastatic disease in bone.

A Monte Carlo simulation model for radiation dose to metastatic skeletal tumor from rhenium-186(Sn)-HEDP.

UNLABELLED: A Monte Carlo model has been developed for simulation of dose delivery to skeletal metastases by the bone surface-seeking radiopharmaceutical 186Re (Sn)-HEDP. METHODS: The model simulates: (1) the heterogeneous small scale geometry of the soft tissue/bone-spicule structure in the lesions as determined by histomorphometric measurements of histologic specimens, (2) the small scale spatial distribution of the radiopharmaceutical on the lesion bone spicule surface as determined by autoradiography, and (3) the 186Re beta and conversion electron decay spectrum and the associated charged particle transport within the modeled geometries. The results are compared with the commonly employed uniform lesion model, which assumes: (1) homogeneous lesion morphology, (2) uniform distribution of radioactivity within the lesion, and (3) complete energy deposition by charged particles within the lesion due to decay of this activity. Gamma and x-ray photons from the 186Re spectrum were assumed to escape from the lesion volume in both models. RESULTS: Results show a significant dependence on the bone volume fraction and hence on the histology of the lesion (lytic, blastic or mixed). The uniform lesion model calculations underestimate the radiation dose to blastic lesions by as much as a factor of 1.8. However, for lytic lesions with low bone volume fractions, both models provide similar dose values. CONCLUSIONS: These new model calculations provide a mechanism for optimizing treatment planning and dose response evaluations of therapeutic bone-seeking radiopharmaceuticals.

Antibody-targeted photolysis of bacteria in vivo.

We have evaluated the efficacy of antibody-targeted photolysis to kill bacteria in vivo using specific antibacterial photosensitizer (PS) immunconjugates. After infecting the dorsal skin in mice with Pseudomonas aeruginosa, both specific and nonspecific tin (IV) chlorin e6-monoclonal antibody conjugates were injected at the infection site. After a 15 min incubation period, the site was exposed to 630 nm light with a power density of 100 mW/cm2 for 1600 seconds. Irradiation resulted in a greater then 75% decrease in the number of viable bacteria at sites treated with a specific conjugate, whereas normal bacterial growth was observed in animals that were untreated or treated with a nonspecific conjugate. Antibody-targeted photolysis may be a selective and versatile tool for treating a variety of infections.

Efficacy of Listerine, Meridol and chlorhexidine mouthrinses as supplements to regular tooth cleaning measures.

The anti-plaque, anti-gingivitis and anti-microbial efficacies of a phenolic compound (Listerine) and 2 different amine/stannous fluoride mouthwashes (Meridol I, II) were compared when these solutions were used in addition to usual tooth cleaning. A placebo preparation was utilized as a negative control and a chlorhexidine solution as a positive control in this double-blind study. After professional tooth cleaning, 49 volunteers continued their habitual, self-performed and non-supervised oral hygiene for a period of 2 weeks, in order to have a more standard baseline. At day 0, they began to rinse twice daily with 1 of the 5 mouthwashes. After 3 weeks of rinsing, plaque indices remained the lowest in the chlorhexidine and the Meridol I groups, while subjects using Listerine or Meridol II demonstrated similar indices significantly lower than that of individuals rinsing with the placebo solution. Through this period, the gingival index scores were similar in the Meridol, Listerine and chlorhexidine groups. At day 21, the mean GI scores in the chlorhexidine group were significantly lower than the scores in the placebo group. The plaque vitality scores showed a bacterial effect in vivo of chlorhexidine and, to a lesser extent, of the Meridol solutions. No substantial evidence of an antibacterial effect in vivo was found for Listerine. This study has demonstrated that when mouthrinses are used to supplement habitual mechanical oral hygiene, chlorhexidine remains the most powerful solution. Furthermore, it was also shown that a combination of habitual self-performed and non-supervised oral hygiene with Meridol or Listerine is more beneficial for plaque control than the use of mechanical oral hygiene alone.

Treatment with tin prevents the development of hypertension in spontaneously hypertensive rats.

Cytochrome P-450-dependent metabolites of arachidonic acid (AA) increased in the kidneys of young, spontaneously hypertensive rats (SHRs) during the period of rapid elevation of blood pressure (BP) but not in adult SHRs or in Wistar Kyoto rats (WKYs) with normal BP. Treatment of SHRs and WKYs with stannous chloride (SnCl2), which selectively depletes renal cytochrome P-450, restored BP to normal, coincident with a natriuresis, in young but not in adult SHRs and did not affect either BP or sodium excretion in WKYs. Depletion of renal cytochrome P-450 was associated with decreased generation of these AA metabolites only in young SHRs. The antihypertensive effect of SnCl2 in young SHRs was greatly reduced by prevention of its cytochrome P-450-depleting action.

Manipulation of cytochrome P-450 dependent renal thromboxane synthase activity in spontaneously hypertensive rats.

Thromboxane synthase is a cytochrome P-450-like enzyme requiring an iron-centered oxygen attack of the prostaglandin endoperoxide substrate (PGH2) for subsequent thromboxane A2 (TxA2) formation. The activity and levels of P-450 enzymes can be manipulated by decreasing heme availability. Stannous chloride (SnCl2) selectively induces renal heme oxygenase activity, depleting heme and decreasing hemoprotein synthesis. We therefore manipulated the renal cytochrome P-450 system to influence thromboxane synthase activity, as measured by the conversion of 14C-PGH2 to thromboxane B2 (TxB2) in renal cortical microsomes from spontaneously hypertensive rats (SHR). Seven-week-old SHR were treated subcutaneously with SnCl2 (1, 10 and 15 mg/100 g body weight) for 4 consecutive days, and cortical microsomal heme oxygenase activity, heme content, P-450 content, thromboxane synthase activity and systolic blood pressure were measured. Heme oxygenase activity was significantly increased from 1058 +/- 62 nmol/mg protein in controls to 3125 +/- 918, 5057 +/- 690--and 4236 +/- 581 nmol/mg protein in SHR treated with 1, 10 and 15 mg/100 g body weight SnCl2, respectively. The increase in heme oxygenase activity was associated with corresponding decreases in heme content (0.29 mumol/mg protein, for control to 0.12 mumol/mg protein for SHR treated with SnCl2, 10 mg/100 g body weight) and cytochrome P-450 content (0.18 +/- 0.1 nmol/mg protein for control to 0.06 +/- 0.01 nmol/mg protein for SHR treated with SnCl2 10 mg/100 g body weight). The reduction in heme and P-450 content was associated with a reduction in thromboxane synthase activity, i.e., decreases of 38, 35 and 47% from control levels at doses of 1, 10 and 15 mg/100 g body weight.(ABSTRACT TRUNCATED AT 250 WORDS)

Tin--a toxic heavy metal? A review of the literature.

A tolerable limit for tin concentration in canned food of 250 ppm (Fritsch et al., 1977) is generally accepted. However, biochemical effects attributable to tin have been observed even after oral administration of 1 and 3 mg Sn/kg body wt (Yamaguchi et al., 1980). These doses reflect 10 and 30 ppm tin in the diet. The experiments of de Groot (1973) showed that hemoglobin concentrations in the blood of rats decreased significantly feeding a diet containing 150 ppm tin. The absorption of iron was diminished after simultaneous administration of 0.8 mumol Sn(II) and iron, reflecting a tin dose of 95 ppm tin, by injection into jejunal loops of rats (Schäfer and Forth, 1983). In general, however, canned food usually plays a secondary role in daily nutrition. Fortunately, concentrations of about 2000 ppm tin as reported by Warburton et al. (1962) and Barker and Runte (1972) are not found in canned food, but values between 50 and 500 ppm are not unusual (Piscator, 1979). If a large amount of canned food is eaten daily over a long period, disturbances of gastric acid secretion and a reduction in iron absorption or heme metabolism cannot be excluded. The storage of food, especially acid foods, in opened cans should be avoided as this practice increases the amount of tin in the food when it is consumed.