From: Subject: Cooperative Effect of Radioimmunotherapy and Antiangiogenic Therapy with Thalidomide in Human Cancer Xenografts -- Kinuya et al. 43 (8): 1084 -- The Journal of Nuclear Medicine Date: Sun, 17 Feb 2008 19:14:05 +0100 MIME-Version: 1.0 Content-Type: multipart/related; type="text/html"; boundary="----=_NextPart_000_0017_01C87199.43379430" X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3198 This is a multi-part message in MIME format. ------=_NextPart_000_0017_01C87199.43379430 Content-Type: text/html; charset="Windows-1252" Content-Transfer-Encoding: quoted-printable Content-Location: http://jnm.snmjournals.org/cgi/content/full/43/8/1084?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&fulltext=glioblastoma&searchid=1&FIRSTINDEX=0&volume=43&issue=8&resourcetype=HWCIT Cooperative Effect of Radioimmunotherapy and = Antiangiogenic Therapy with Thalidomide in Human Cancer Xenografts -- = Kinuya et al. 43 (8): 1084 -- The Journal of Nuclear Medicine
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Journal of Nuclear Medicine Vol. 43 No. 8 1084-1089
=A9 = 2002 by Society of Nuclear=20 Medicine

Basic Science Investigations

Cooperative Effect of Radioimmunotherapy and Antiangiogenic Therapy = with=20 Thalidomide in Human Cancer Xenografts

Seigo = Kinuya,=20 MD1, Atsuhiro Kawashima, MD2,=20 Kunihiko Yokoyama, MD1, Kiyoshi Koshida,=20 MD3, Shota Konishi, MD1, = Naoto=20 Watanabe, MD4, Noriyuki Shuke, = MD5,=20 Hisashi Bunko, MD6, Takatoshi Michigishi,=20 MD1 and Norihisa Tonami, MD1 =

1 Department of Biotracer Medicine, = Kanazawa=20 University Graduate School of Medical Sciences, Kanazawa, = Japan
2=20 Department of Molecular and Cellular Pathology, Kanazawa University = Graduate=20 School of Medical Sciences, Kanazawa, Japan
3 Department = of=20 Integrative Cancer Therapy and Urology, Kanazawa University Graduate = School of=20 Medical Sciences, Kanazawa, Japan
4 Department of = Radiology,=20 Toyama Medical and Pharmaceutical University, Toyama, = Japan
5=20 Department of Radiology, Asahikawa Medical College, Asahikawa,=20 Japan
6 Medical Informatics, Kanazawa University Hospital, = Kanazawa, Japan


3D"   =20 ABSTRACT
TOP
3D"ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
CONCLUSION
REFERENCES
 
A= ntiangiogenic=20 therapy may prolong the dormancy of cancer lesions. Moreover, = radioimmunotherapy (RIT) may eradicate this population of = cells. This=20 study dealt with determining the benefits associated with the = combined usefulness of these 2 therapies with respect to = inhibition=20 of tumor growth. Methods: Antiangiogenic therapy using = oral=20 thalidomide (daily dose, 200 mg/kg) and RIT involving a = single=20 intravenous injection (4.63 MBq 131I-A7, an IgG1 murine=20 monoclonal antibody) were conducted in mice bearing LS180 = human=20 colon cancer xenografts. RIT with an irrelevant IgG1, HPMS-1, = was also performed as a control. Antiangiogenesis of = thalidomide=20 was investigated by immunohistochemical analysis of tumor = sections.=20 Results: Antiangiogenic therapy and RIT with = 131I-A7=20 significantly suppressed the growth of xenografts. This = combination=20 produced more efficient tumor growth inhibition than did the=20 monotherapy (P < 0.005). RIT using = 131I-HPMS-1=20 was far less effective than 131I-A7, even when = combined=20 with thalidomide administration. Immunohistochemistry = revealed a=20 decrease in the microvessel number within tumors treated with = thalidomide (P < 0.0001). Combined therapy further = reduced=20 the microvessel number (P < 0.01 vs. thalidomide=20 monotherapy). Conclusion: The combination of RIT and=20 thalidomide antiangiogenic therapy produces a better response = of=20 tumors than does monotherapy. Acting in concert, = antiangiogenic=20 therapy may prolong the dormancy of cancer lesions and RIT = may=20 eradicate this population of cells.

Key Words: radioimmunotherapy =95 antiangiogenic = therapy =95=20 thalidomide


3D"   =20 INTRODUCTION
TOP
ABSTRACT
3D"INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
CONCLUSION
REFERENCES
 
T= heoretically,=20 radioimmunotherapy (RIT) may selectively treat cancer = lesions;=20 moreover, systemic administration of radiolabeled monoclonal = antibody=20 (mAb) may enable the treatment of disseminated metastatic = lesions.=20 However, results reported for conventional RIT in most solid = tumors,=20 with the exception of malignant lymphoma, have not been = favorable (1).=20 Consequently, modification of RIT is considered to effect = killing of=20 solid tumors (1).=20

Tumor growth depends on neoangiogenesis; therefore, = antiangiogenic=20 therapy using agents that suppress endothelial cell growth = has=20 been investigated extensively (2).=20 The objective of this treatment is to maintain lesions at = limited=20 size, a situation referred to as tumor dormancy (2).=20 We hypothesize that the combined usefulness of antiangiogenic = therapy=20 and RIT may synergistically effect treatment of cancer = lesions. The=20 rationale for this hypothesis is based on the suppression of = tumor=20 endothelial compartments by antiangiogenic agents as well as = the=20 direct action of RIT against tumor cells. In addition, = antiangiogenic=20 therapy induces dormancy of lesions and RIT may eradicate = this=20 population of dormant cells.

Thalidomide was developed as a sedative; however, it was = subsequently=20 withdrawn from clinical use because of its teratogenicity. = This=20 agent is now known to exert effects on the immune system. = Thalidomide=20 inhibits the production of tumor necrosis factor-3D{alpha}=20, = stimulates=20 cytotoxic T-cell proliferation, and induces secretion of = interferon-=20 and interleukin-2 (3,4).=20 On the basis of its antiinflammatory properties, thalidomide = is used=20 in the treatment of such diseases as erythema nodosum = leprosum and=20 graft-versus-host disease. In addition, thalidomide possesses = potent=20 antiangiogenic properties (5=967),=20 which arise as a result of blockade of the action of = angiogenic=20 factors such as vascular endothelial growth factor (VEGF) (8)=20 and basic fibroblast growth factor (bFGF) (5).=20 Consequently, this agent is being investigated currently in = clinical=20 trials of patients exhibiting various types of malignancies = (3,4).=20 This study dealt with determining the effects of combined RIT = and thalidomide therapy in the treatment of xenografts of = human=20 colon cancer cells.


3D"   =20 MATERIALS AND METHODS
TOP
ABSTRACT
INTRODUCTION
3D"MATERIALS AND METHODS
RESULTS
DISCUSSION
CONCLUSION
REFERENCES
 
<= STRONG>mAb=20 and Tumor Model
A7, an IgG1 murine mAb with 3D{kappa}=20-light chain, = was used in=20 this investigation (9).=20 It was purified from ascites of hybridoma-bearing mice by = protein=20 A-Sepharose column (Bio-Rad, Richmond, CA) chromatography. A7 = mAb=20 recognizes a 45-kDa surface glycoprotein of human colonic = carcinomas.=20 An additional IgG1 recognizing placental alkaline = phosphatase,=20 HPMS-1, was used as a class-matched irrelevant control mAb = (10).=20 The mAbs were radiolabeled with 131I by the = chloramine-T=20 method and subsequently purified on a PD-10 column (Pharmacia = LKB=20 Biotechnology, Uppsala, Sweden). The specific activity of = purified=20 131I-A7 was 142 MBq/mg. Immunoreactivity of = purified=20 131I-A7 was assessed as 69% in antigen excess conditions = with an LS180 human colon cancer cell line. The specific = activity=20 of 131I-HPMS-1 was 90 MBq/mg. Antibodies were = sterilized=20 by filtration (Millex-GV, 0.22 mm; Millipore, Bedford, MA)=20 before further experiments. Animal studies were performed in=20 compliance with the regulations of our institutions, and the=20 principles of laboratory animal care were followed. BALB/c=20 nu/nu mice (female, 6=968 wk old) were xenografted=20 subcutaneously in the thigh area with 5 x 106 LS180 cells. Animals were = subjected=20 to treatment when xenografts attained a diameter of = approximately=20 5=967 mm. This cell line reportedly shows considerable VEGF=20 production; moreover, the growth of xenografts in mice is = suppressed=20 by an anti-VEGF antibody (11).=20

Antitumor Effect of 131I-A7 with=20 Thalidomide
Tumor volume (mm3), calculated as = length (mm)=20 x width (mm)2 = x 0.5, was expressed as a ratio of volumes = to the=20 volume obtained on the initial day of treatment (day 0). = Tumor volume=20 on day 0 was measured as 139 =B1 21 mm3. The = therapeutic=20 experiment consisted of 6 groups: nontreated mice (n = =3D 5);=20 mice receiving a daily oral dose of 200 mg/kg thalidomide = (n =3D=20 10); mice undergoing treatment with 4.63 MBq = 131I-A7=20 (n =3D 10); mice treated with 131I-A7 and a = daily=20 dose of thalidomide (n =3D 10); mice treated with 4.63 = MBq of an=20 irrelevant mAb, 131I-HPMS-1 (n =3D 5); and = mice=20 receiving 131I-HPMS-1 and a daily dose of thalidomide = (n=20 =3D 6). Thalidomide suspensions were prepared in 0.1 mL 0.5%=20 carboxymethylcellulose (Wako, Osaka, Japan) and administered = orally.=20 The dose of 131I-A7 used in this study was = approximately=20 half of the maximum tolerated dose in this model. Thalidomide = dosage=20 was determined in accordance with previous reports (5=967).=20 This dosage has not been reported to cause critical side = effects.=20 Nontreated mice and mice receiving RIT monotherapy were = orally=20 administered an identical volume of carboxymethylcellulose = that did=20 not contain thalidomide. The therapeutic efficacy of each = treatment=20 was analyzed statistically by tumor size on day 13. = Furthermore,=20 analysis with the area under the curve (AUC) of tumor growth = curves,=20 which would be the reflection of the therapeutic effect = during the=20 entire observation period, was also performed for the = comparison=20 among the treatment groups.

Toxicity of Treatment
A major critical organ in = RIT is=20 the bone marrow. Thalidomide may cause neutropenia (3).=20 Therefore, we attempted to establish whether the combined = therapy=20 increased myelotoxicity in animals. Nontumor-bearing mice = were=20 partitioned into 4 groups: nontreated mice, mice receiving = RIT (4.63=20 MBq), mice treated with a daily dose of thalidomide (200 = mg/kg), and=20 mice undergoing the combined regimen (n =3D 3). = Thalidomide=20 administration was conducted until day 23. A blood sample (5 = =B5L) was=20 obtained from a tail vein in each mouse. Samples from mice = within a=20 single group were pooled and diluted 1:10,000 in = phosphate-buffered=20 saline for red blood cell (RBC) counts, 1:20 in 3% acetic = acid=20 for white blood cell (WBC) counts, and 1:100 in 1% ammonium=20 oxalate for platelet counts (12).=20 Body weight of the animals was also determined.

Assessment of Microvessel Density in = Tumors
Tumors were=20 excised from nontreated control mice, mice treated with = either=20 thalidomide or 131I-A7 RIT monotherapy, and mice = treated=20 with the combined therapy on day 15. Specimens were fixed = with 10%=20 formalin and embedded in paraffin wax (n =3D 3).=20 Immunoperoxidase staining using a rabbit polyclonal antibody=20 against factor VIII (DAKO, Copenhagen, Denmark) was = performed.=20 The microvessel density was determined by light microscopy = according=20 to the procedure described by Weidner et al. (13).=20 Briefly, the areas of highest vascularization were detected = by=20 low-power scanning (100x) = of the=20 tumor sections. Individual microvessels were then counted in = a=20 200x field, with 5 different fields=20 examined in each section. Two sections per tumor were = studied, and=20 the average of 3 tumors in each group was compared. =

In Vitro Assay with Thalidomide
Cells were = incubated in=20 24-well culture plates at 20,000 cells per milliliter in = Dulbecco=92s=20 modified Eagle medium (Nissui Seiyaku, Tokyo, Japan) to = determine=20 whether thalidomide would exert a direct effect on LS180 cell = growth.=20 Increasing concentrations of thalidomide (up to 100 =B5mol/L) = were=20 introduced. The agents were dissolved in dimethyl sulfoxide = ([DMSO];=20 Wako, Tokyo, Japan) and diluted in the medium. The final=20 concentration of DMSO was 0.5%. Control cells were treated = with DMSO=20 alone. Before the assay, confirmation was obtained that DMSO = at this=20 concentration did not affect proliferation of LS180 cells. = Cells were=20 exposed to trypsin, and the viable cell number was then = determined=20 1, 3, and 5 d after inoculation (n =3D 3).

The possibility of radiosensitization effects of thalidomide = on=20 LS180 cells was investigated. Briefly, cells were treated = with=20 thalidomide at various concentrations up to 100 =B5mol/L for = 24 h,=20 followed by exposure to an x-ray beam (0=968 Gy) (MBR-1520R; = Hitachi,=20 Tokyo, Japan) at 1.41 Gy/min. Cell proliferation was = observed, and=20 the surviving fraction was calculated as described (14).=20

Statistical Analysis
Results were analyzed by = 1-way ANOVA=20 with Fisher=92s protected least-significant difference. The = level of=20 significance was set at 5% .


3D"   =20 RESULTS
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
3D"RESULTS
DISCUSSION
CONCLUSION
REFERENCES
 
E= ither=20 thalidomide or 131I-A7 RIT monotherapy significantly=20 suppressed growth of LS180 xenografts in comparison with no=20 treatment (P < 0.005 and P < 0.0005, = respectively) (Fig.=20 1). Relative tumor volumes on day 13 were 12.7 =B1 = 2.95,=20 6.84 =B1 0.71, and 3.05 =B1 0.37 in the control group, = the group=20 administered thalidomide, and the group receiving = 131I-A7=20 RIT, respectively. Treatment with a class-matched irrelevant = mAb,=20 131I-HPMS-1, did not effectively suppress tumor growth=20 (P =3D 0.06). The combined treatment of thalidomide and=20 131I-A7 RIT resulted in more efficient tumor = growth=20 inhibition than did monotherapy with thalidomide or RIT = alone,=20 displaying a tumor volume of 1.31 =B1 0.33 on day 13 = (P <=20 0.005 vs. RIT alone). Moreover, 6 of 10 tumors regressed with = combined treatment, whereas no regression occurred with=20 monotherapy. Results of AUC analysis of tumor growth curves = also=20 indicated the superiority of the combined regimen of = thalidomide and=20 131I-A7 RIT over monotherapy for the inhibition of = tumor=20 growth (Table=20 1). The combination of 131I-HPMS-1 and = thalidomide was=20 far less effective than the combination of 131I-A7 = and=20 thalidomide.



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  		FIGURE 1. Growth of LS180 = colon=20 cancer xenografts. Tumor volume is expressed as ratio = relative to=20 volume on day 0. 3D{diamondsuit}=20,=20 Nontreated control; 3D{blacktriangleup}=20, tumors=20 treated with 200 mg/kg/d thalidomide; 3D{square}=20, tumors=20 treated with 4.63 MBq 131I-HPMS-1; 3D{blacksquare}=20, tumors=20 treated with thalidomide and 131I-HPMS-1; 3D{circ}=20, = tumors=20 treated with 4.63 MBq 131I-A7; =95, tumors = treated with=20 thalidomide and 131I-A7. =

 = ;

View = this=20 table:
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  		TABLE 1 Analysis of AUC = of Growth=20 Curves of Colon Cancer Xenografts After Initiation of = Treatment =

 = ;
Thalidomide=20 treatment induced minor depression of the peripheral WBC = count=20 relative to that observed in nontreated mice (Fig.=20 2). The nadir of WBC count depression in mice treated = with=20 the combination of RIT and thalidomide therapy was similar to = that in mice treated with RIT alone; however, the depression=20 persisted in mice treated with the combination regimen. The = WBC=20 count recovered after the cessation of thalidomide=20 administration. Changes in the platelet count in mice = receiving the=20 combination treatment did not vary from those in mice = undergoing RIT=20 alone. Thalidomide administration did not affect the RBC = count.=20 The use of thalidomide did not result in weight loss that was = different than that of the control (Fig.=20 3). The maximal body weight loss was similar in mice = treated with=20 RIT alone and those undergoing the combination regimen; = however,=20 recovery was delayed by thalidomide treatment. No mice showed = signs=20 of diarrhea or infection.



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  		FIGURE 2. Changes in = peripheral=20 blood cell counts in mice. (A) RBCs; (B) WBCs; (C) = platelets. , Nontreated control; 3D{blacktriangleup}=20, tumors=20 treated with 200 mg/kg/d thalidomide; 3D{circ}=20, = tumors=20 treated with 4.63-MBq RIT; =95, tumors treated with = thalidomide and=20 RIT. =

 = ;


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  		FIGURE 3. Body weight = changes of=20 mice. 3D{diamondsuit}=20,=20 Nontreated control; 3D{blacktriangleup}=20, tumors=20 treated with 200 mg/kg/d thalidomide; 3D{circ}=20, = tumors=20 treated with 4.63-MBq RIT; =95, tumors treated with = thalidomide and=20 RIT. =

 = ;
Immunostaining=20 of tumor sections with antifactor VIII antibody showed = significant=20 reduction in the microvessel density in tumors treated with=20 thalidomide (8.30 =B1 0.8 per 200x = field)=20 in comparison with nontreated control tumors (29.9 =B1 = 2.5)=20 (P < 0.0001) (Fig.=20 4). Interestingly, the microvessel density in tumors = treated with=20 131I-A7 RIT (10.8 =B1 1.4) was lower than that in = control=20 tumors (P < 0.0001). Moreover, the combination of = RIT and=20 thalidomide treatment further decreased the microvessel = density in=20 tumors (4.3 =B1 0.5; P < 0.0001 vs. RIT monotherapy = and=20 P < 0.01 vs. thalidomide monotherapy).



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  		FIGURE 4. = Immunohistochemistry of=20 tumor sections with an antifactor VIII antibody. Microvessel = numbers=20 in 200x field are 29.9 = =B1 2.5 in=20 control tumor (A) and 8.30 =B1 0.8 in tumor treated with = thalidomide=20 (B) (P < 0.0001). =

 = ;
The=20 in vitro proliferation assay revealed no evidence of direct=20 cytotoxicity of thalidomide in LS180 cells up to = concentrations=20 of 100 =B5mol/L (Fig.=20 5). In addition, thalidomide did not radiosensitize the = cells (Fig.=20 6).



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  		FIGURE 5. In vitro = proliferation=20 assay of LS180 colon cancer cells in presence of increasing=20 concentrations of thalidomide (n =3D 3). 3D{blacksquare}=20, Without=20 thalidomide; in presence of thalidomide: =95, 0.1 =B5mol/L; = , 1 =B5mol/L;=20 3D{diamondsuit}=20, 5 =B5mol/L;=20 3D{square}, 10 =B5 mol/L; 3D{circ}=20, = 50 =B5mol/L;=20 3D{triangleup}, 100 =B5mol/L. =

 = ;


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  		FIGURE 6. Clonogenic = assay of LS180=20 colon cancer cells exposed to x-ray radiation (n =3D = 3). , Control cells; cells pretreated with = thalidomide for 24=20 h: 3D{blacksquare}=20, 1 =B5mol/L;=20 3D{blacktriangleup}=20, 10=20 =B5mol/L; 3D""=20, 100=20 =B5mol/L. =

 = ;

3D"   =20 DISCUSSION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
3D"DISCUSSION
CONCLUSION
REFERENCES
 
T= halidomide=20 is currently being investigated in several clinical trials = involving=20 tumors such as low-grade glioma, AIDS-related Kaposi=92s = sarcoma, renal=20 cell carcinoma, hepatocellular carcinoma, and multiple = myeloma (3).=20 The effectiveness of thalidomide in colorectal cancer = patients is=20 also currently being evaluated (3).=20 It is known that the degree of tumor vascularization is a = prognostic=20 factor of colon cancer and is correlated with its metastases = and=20 recurrence (15).=20 Expression of VEGF and bFGF also correlates with prognosis in = colon=20 cancer patients (16).=20 A neutralizing anti-VEGF mAb exhibits antitumor activity in=20 xenografts of various colon cancer cell lines (11).=20 In our study, the tumor-suppressive effects of thalidomide = were=20 detected in xenografts of LS180 colon cancer cells. These = facts=20 suggest a role for antiangiogenic therapy in patients with = colon=20 cancer. However, because the response rates of solid tumors = to=20 monotherapy with thalidomide are limited in clinical trials, = the=20 combination with chemotherapy or radiotherapy has been = considered.=20 The efficacy of conventional RIT is also currently limited in = most=20 types of tumors; consequently, several methods have been=20 proposed to improve its therapeutic outcome (1).=20

The combination of antiangiogenic therapy and RIT proposed in = this=20 investigation is a novel approach in which 2 agents act = against=20 distinct compartments within tumors: Antiangiogenic agents = suppress=20 the endothelial compartment, whereas RIT acts directly = against the=20 tumor cell. We showed that thalidomide was unable to exert = direct=20 cytotoxicity on LS180 cells or to sensitize cells to = radiation;=20 however, thalidomide reduced the microvessel density in = xenografts in=20 mice. These results clearly indicate that the cooperative = improvement=20 of tumor growth suppression with combined therapy was related = to this=20 hypothesis. The effectiveness of combining RIT and another=20 antiangiogenic agent, 2-methoxyestradiol, has been reported = recently=20 (17).=20

Interestingly, 131I-A7 RIT monotherapy reduced the=20 microvessel number in tumors compared with no treatment; = furthermore,=20 the combined regimen more effectively decreased the = microvessel=20 number than did thalidomide monotherapy. These findings were=20 probably caused by radiation injury of the vessels due to=20 131I-A7. Because penetration of extravasated mAbs = into=20 solid tumors is disturbed by factors such as high = interstitial=20 pressure (18)=20 and the so-called binding site barrier (19),=20 mAbs would deposit close to the vasculature; for instance, = IgG would=20 require days to cover the distance of 3D~=201 mm after = the=20 extravasation (18).=20 Consequently, considerable radiation dose would be absorbed = by=20 the vascular compartment. In this sense, RIT and antiangiogenic=20 therapy would cooperatively give effect to suppression of the = tumor vasculature in the combined regimen.

Changes in peripheral blood cell counts and the body weight = of=20 mice were monitored to assess the toxicity of treatment. In = addition=20 to teratogenicity, thalidomide displays several adverse = effects,=20 including somnolence, constipation, skin rashes, orthostatic=20 hypotension, nausea, peripheral neuropathy, and neutropenia = (3,=20 4).=20 Most of these untoward effects do not occur in RIT therapy; = however,=20 neutropenia may affect combination with RIT. Minor depression = of the=20 peripheral WBC count was observed in mice that were treated = daily=20 with thalidomide. The combined regimen delayed recovery of = the WBC=20 count in comparison with RIT monotherapy. Recovery of body = weight=20 loss was also delayed by this combination. Reports describing = clinical trials of thalidomide indicate that = thalidomide-induced=20 neutropenia occurs rarely in patients other than those = presenting=20 with HIV infection and recipients of an allogeneic bone = marrow=20 transplant (3,4);=20 however, marrow toxicity should be carefully monitored during = combination therapy involving RIT and thalidomide. =

Clinical results of RIT have been disappointing in the treatment = of bulky disease of solid tumors (1).=20 However, several reports have described the role of RIT in an = adjuvant setting of cancer treatment, in which small residual = tumors=20 are targeted by mAbs (20=9623).=20 Antiangiogenic therapy also has an important role in this = setting, in=20 which angiogenesis inhibitors may induce dormancy of = microscopic=20 metastases or stabilization of residual disease (24=9626).=20 Because antiangiogenic therapy alone may not produce a = complete cure,=20 a combination with cytotoxic therapy is required to be = curative (1).=20 In this regard, a combined regimen with external beam = radiotherapy=20 has been proven to enhance tumor growth suppression in = various kinds=20 of tumors (27,28).=20 However, as a local treatment, external beam radiotherapy = should=20 have a limitation in patients with metastatic foci. The = combined=20 regimen with RIT and antiangiogenic therapy surely has an = advantage=20 in this setting, because both treatments can be administered=20 systemically.


3D"   =20 CONCLUSION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
3D"CONCLUSION
REFERENCES
 
W= e=20 observed a cooperative relationship in the combined usefulness = of RIT=20 and antiangiogenic therapy with thalidomide in the treatment = of colon=20 cancer xenografts. Acting in concert, antiangiogenic therapy = may=20 prolong the dormancy of cancer lesions and RIT may eradicate = this=20 population of cells. We are currently testing this hypothesis = in=20 liver metastasis and intraperitoneal metastasis models. =


3D"   =20 ACKNOWLEDGMENTS
 
The = authors thank=20 former Professor Toshio Takahashi and Dr. Toshiharu Yamaguchi = (First=20 Department of Surgery, Kyoto Prefectural University of = Medicine) for=20 providing A7 mAb. This study was supported in part by = grants-in-aid=20 from the Ministry of Education, Science, Sports and Culture,=20 Japan.


3D"   =20 FOOTNOTES
 
Received Oct. 29, 2001; revision accepted = Apr. 19,=20 2002.

For correspondence or reprints contact: Seigo Kinuya, MD, = Department=20 of Biotracer Medicine, Kanazawa University Graduate School of = Medical Sciences, 13-1 Takaramachi, Kanazawa, Ishikawa = 920-8640,=20 Japan.

E-mail: kinuya{at}med.kanazawa-u.ac.jp


3D"   =20 REFERENCES
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
CONCLUSION
3D"REFERENCES
&nb= sp;

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Opera = requires=0A= the Sun Java Runtime Environment .=0A= =0A= by Andrew Gregory=0A= http://www.scss.com.au/family/andrew/webdesign/xmlhttprequest/=0A= =0A= This work is licensed under the Creative Commons Attribution License. To = view a=0A= copy of this license, visit = http://creativecommons.org/licenses/by-sa/2.5/ or=0A= send a letter to Creative Commons, 559 Nathan Abbott Way, Stanford, = California=0A= 94305, USA.=0A= =0A= Attribution: Leave my name and web address in this script intact.=0A= =0A= Not Supported in Opera=0A= ----------------------=0A= * user/password authentication=0A= * responseXML data member=0A= =0A= Not Fully Supported in Opera=0A= ----------------------------=0A= * async requests=0A= * abort()=0A= * getAllResponseHeaders(), getAllResponseHeader(header)=0A= =0A= */=0A= // IE support=0A= if (window.ActiveXObject && !window.XMLHttpRequest) {=0A= window.XMLHttpRequest =3D function() {=0A= var msxmls =3D new Array(=0A= 'Msxml2.XMLHTTP.5.0',=0A= 'Msxml2.XMLHTTP.4.0',=0A= 'Msxml2.XMLHTTP.3.0',=0A= 'Msxml2.XMLHTTP',=0A= 'Microsoft.XMLHTTP');=0A= for (var i =3D 0; i < msxmls.length; i++) {=0A= try {=0A= return new ActiveXObject(msxmls[i]);=0A= } catch (e) {=0A= }=0A= }=0A= return null;=0A= };=0A= }=0A= // Gecko support=0A= /* ;-) */=0A= // Opera support=0A= if (window.opera && !window.XMLHttpRequest) {=0A= window.XMLHttpRequest =3D function() {=0A= this.readyState =3D 0; // = 0=3Duninitialized,1=3Dloading,2=3Dloaded,3=3Dinteractive,4=3Dcomplete=0A= this.status =3D 0; // HTTP status codes=0A= this.statusText =3D '';=0A= this._headers =3D [];=0A= this._aborted =3D false;=0A= this._async =3D true;=0A= this._defaultCharset =3D 'ISO-8859-1';=0A= this._getCharset =3D function() {=0A= var charset =3D _defaultCharset;=0A= var contentType =3D = this.getResponseHeader('Content-type').toUpperCase();=0A= val =3D contentType.indexOf('CHARSET=3D');=0A= if (val !=3D -1) {=0A= charset =3D contentType.substring(val);=0A= }=0A= val =3D charset.indexOf(';');=0A= if (val !=3D -1) {=0A= charset =3D charset.substring(0, val);=0A= }=0A= val =3D charset.indexOf(',');=0A= if (val !=3D -1) {=0A= charset =3D charset.substring(0, val);=0A= }=0A= return charset;=0A= };=0A= this.abort =3D function() {=0A= this._aborted =3D true;=0A= };=0A= this.getAllResponseHeaders =3D function() {=0A= return this.getAllResponseHeader('*');=0A= };=0A= this.getAllResponseHeader =3D function(header) {=0A= var ret =3D '';=0A= for (var i =3D 0; i < this._headers.length; i++) {=0A= if (header =3D=3D '*' || this._headers[i].h =3D=3D header) {=0A= ret +=3D this._headers[i].h + ': ' + this._headers[i].v + '\n';=0A= }=0A= }=0A= return ret;=0A= };=0A= this.getResponseHeader =3D function(header) {=0A= var ret =3D getAllResponseHeader(header);=0A= var i =3D ret.indexOf('\n');=0A= if (i !=3D -1) {=0A= ret =3D ret.substring(0, i);=0A= }=0A= return ret;=0A= };=0A= this.setRequestHeader =3D function(header, value) {=0A= this._headers[this._headers.length] =3D {h:header, v:value};=0A= };=0A= this.open =3D function(method, url, async, user, password) {=0A= this.method =3D method;=0A= this.url =3D url;=0A= this._async =3D true;=0A= this._aborted =3D false;=0A= this._headers =3D [];=0A= if (arguments.length >=3D 3) {=0A= this._async =3D async;=0A= }=0A= if (arguments.length > 3) {=0A= opera.postError('XMLHttpRequest.open() - user/password not = supported');=0A= }=0A= this.readyState =3D 1;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= };=0A= this.send =3D function(data) {=0A= if (!navigator.javaEnabled()) {=0A= alert("XMLHttpRequest.send() - Java must be installed and = enabled.");=0A= return;=0A= }=0A= if (this._async) {=0A= setTimeout(this._sendasync, 0, this, data);=0A= // this is not really asynchronous and won't execute until the = current=0A= // execution context ends=0A= } else {=0A= this._sendsync(data);=0A= }=0A= }=0A= this._sendasync =3D function(req, data) {=0A= if (!req._aborted) {=0A= req._sendsync(data);=0A= }=0A= };=0A= this._sendsync =3D function(data) {=0A= this.readyState =3D 2;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= // open connection=0A= var url =3D new java.net.URL(new = java.net.URL(window.location.href), this.url);=0A= var conn =3D url.openConnection();=0A= for (var i =3D 0; i < this._headers.length; i++) {=0A= conn.setRequestProperty(this._headers[i].h, this._headers[i].v);=0A= }=0A= this._headers =3D [];=0A= if (this.method =3D=3D 'POST') {=0A= // POST data=0A= conn.setDoOutput(true);=0A= var wr =3D new = java.io.OutputStreamWriter(conn.getOutputStream(), this._getCharset());=0A= wr.write(data);=0A= wr.flush();=0A= wr.close();=0A= }=0A= // read response headers=0A= // NOTE: the getHeaderField() methods always return nulls for me :(=0A= var gotContentEncoding =3D false;=0A= var gotContentLength =3D false;=0A= var gotContentType =3D false;=0A= var gotDate =3D false;=0A= var gotExpiration =3D false;=0A= var gotLastModified =3D false;=0A= for (var i =3D 0; ; i++) {=0A= var hdrName =3D conn.getHeaderFieldKey(i);=0A= var hdrValue =3D conn.getHeaderField(i);=0A= if (hdrName =3D=3D null && hdrValue =3D=3D null) {=0A= break;=0A= }=0A= if (hdrName !=3D null) {=0A= this._headers[this._headers.length] =3D {h:hdrName, = v:hdrValue};=0A= switch (hdrName.toLowerCase()) {=0A= case 'content-encoding': gotContentEncoding =3D true; break;=0A= case 'content-length' : gotContentLength =3D true; break;=0A= case 'content-type' : gotContentType =3D true; break;=0A= case 'date' : gotDate =3D true; break;=0A= case 'expires' : gotExpiration =3D true; break;=0A= case 'last-modified' : gotLastModified =3D true; break;=0A= }=0A= }=0A= }=0A= // try to fill in any missing header information=0A= var val;=0A= val =3D conn.getContentEncoding();=0A= if (val !=3D null && !gotContentEncoding) = this._headers[this._headers.length] =3D {h:'Content-encoding', v:val};=0A= val =3D conn.getContentLength();=0A= if (val !=3D -1 && !gotContentLength) = this._headers[this._headers.length] =3D {h:'Content-length', v:val};=0A= val =3D conn.getContentType();=0A= if (val !=3D null && !gotContentType) = this._headers[this._headers.length] =3D {h:'Content-type', v:val};=0A= val =3D conn.getDate();=0A= if (val !=3D 0 && !gotDate) this._headers[this._headers.length] = =3D {h:'Date', v:(new Date(val)).toUTCString()};=0A= val =3D conn.getExpiration();=0A= if (val !=3D 0 && !gotExpiration) = this._headers[this._headers.length] =3D {h:'Expires', v:(new = Date(val)).toUTCString()};=0A= val =3D conn.getLastModified();=0A= if (val !=3D 0 && !gotLastModified) = this._headers[this._headers.length] =3D {h:'Last-modified', v:(new = Date(val)).toUTCString()};=0A= // read response data=0A= var reqdata =3D '';=0A= var stream =3D conn.getInputStream();=0A= if (stream) {=0A= var reader =3D new java.io.BufferedReader(new = java.io.InputStreamReader(stream, this._getCharset()));=0A= var line;=0A= while ((line =3D reader.readLine()) !=3D null) {=0A= if (this.readyState =3D=3D 2) {=0A= this.readyState =3D 3;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= }=0A= reqdata +=3D line + '\n';=0A= }=0A= reader.close();=0A= this.status =3D 200;=0A= this.statusText =3D 'OK';=0A= this.responseText =3D reqdata;=0A= this.readyState =3D 4;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= if (this.onload) {=0A= this.onload();=0A= }=0A= } else {=0A= // error=0A= this.status =3D 404;=0A= this.statusText =3D 'Not Found';=0A= this.responseText =3D '';=0A= this.readyState =3D 4;=0A= if (this.onreadystatechange) {=0A= this.onreadystatechange();=0A= }=0A= if (this.onerror) {=0A= this.onerror();=0A= }=0A= }=0A= };=0A= };=0A= }=0A= // ActiveXObject emulation=0A= if (!window.ActiveXObject && window.XMLHttpRequest) {=0A= window.ActiveXObject =3D function(type) {=0A= switch (type.toLowerCase()) {=0A= case 'microsoft.xmlhttp':=0A= case 'msxml2.xmlhttp':=0A= case 'msxml2.xmlhttp.3.0':=0A= case 'msxml2.xmlhttp.4.0':=0A= case 'msxml2.xmlhttp.5.0':=0A= return new XMLHttpRequest();=0A= }=0A= return null;=0A= };=0A= }=0A= ------=_NextPart_000_0017_01C87199.43379430 Content-Type: application/octet-stream Content-Transfer-Encoding: quoted-printable Content-Location: http://jnm.snmjournals.org/javascript/ajax/utility.js /************************************************************************= *****=0A= * javascript/ajax/utility.js=0A= *=0A= * Utility functions for working with XMLHttpRequest data.=0A= *=0A= * Copyright 2006 Board of Trustees of the Leland Stanford Junior = University.=0A= = *************************************************************************= ***/=0A= =0A= /*=0A= * Copy XML nodes into an HTMLElement. This effectively=0A= * clones XML markup which uses XHTML naming conventions=0A= * into an HTML DOM.=0A= */=0A= function copy_xml_to_html(src, dst) {=0A= if (src.nodeType =3D=3D 1) { /* Node.ELEMENT_NODE */=0A= var e =3D document.createElement(src.nodeName);=0A= for (var i =3D 0; i < src.childNodes.length; i++) {=0A= copy_xml_to_html(src.childNodes[i], e);=0A= }=0A= for (var i =3D 0; i < src.attributes.length; i++) {=0A= var n =3D src.attributes[i].name;=0A= var v =3D unescape_xml_string(src.attributes[i].value); =0A= e.setAttribute(n, v);=0A= if (n =3D=3D "class") {=0A= e.className =3D v;=0A= }=0A= else if (n =3D=3D "style") {=0A= set_css_style(v, e, "");=0A= }=0A= }=0A= dst.appendChild(e);=0A= }=0A= else if (src.nodeType =3D=3D 3) { /* Node.TEXT_NODE */=0A= dst.appendChild(document.createTextNode(src.nodeValue));=0A= }=0A= }=0A= =0A= /* =0A= * It is unclear that this is the right thing to be calling=0A= * from copy_xml_to_html, but it appears that Safari decides=0A= * to convert & to the NCR #, and then encodes that=0A= * NCR to &%26%2338;. So, I'm going to treat the DOM Attr=0A= * value as a plain string, and run our XML string input=0A= * through the decoding routine below.=0A= */=0A= function unescape_xml_string(s) {=0A= return s.replace(/'/g, "'")=0A= .replace(/'/g, "'")=0A= .replace(/"/g, "\"")=0A= .replace(/"/g, "\"")=0A= .replace(/>/g, ">")=0A= .replace(/>/g, ">")=0A= .replace(/</g, "<")=0A= .replace(/</g, "<")=0A= .replace(/&/g, "&")=0A= .replace(/&/g, "&");=0A= }=0A= =0A= /*=0A= * Parse set of CSS rules and apply them to an element.=0A= * This is quite horrifying, but I'm unable to determine=0A= * how else to handle this with IE 6. FireFox and other=0A= * sane browsers let you simply set the style attribute=0A= * or use e.style.setProperty(rule, value, priority),=0A= * IE 6 appears to have neither of these capabilities..=0A= */=0A= function set_css_style(css, e, priority) {=0A= var rules =3D css.split(";");=0A= for (var i =3D 0; i < rules.length; i++) {=0A= var nvpair =3D rules[i].split(":");=0A= if (nvpair.length =3D=3D 2) {=0A= try {=0A= var name =3D nvpair[0]; /* style attribute */=0A= var value =3D nvpair[1]; /* attribute value */=0A= =0A= /*=0A= * For each possible style attribute, set the=0A= * appropriate style property in the element.=0A= */=0A= if (name =3D=3D "background") {=0A= e.style.background =3D value;=0A= }=0A= else if (name =3D=3D "background-attachment") {=0A= e.style.backgroundAttachment =3D value;=0A= }=0A= else if (name =3D=3D "background-color") {=0A= e.style.backgroundColor =3D value;=0A= }=0A= else if (name =3D=3D "background-image") {=0A= e.style.backgroundImage =3D value;=0A= }=0A= else if (name =3D=3D "background-position") {=0A= e.style.backgroundPosition =3D value;=0A= }=0A= else if (name =3D=3D "background-position-x") {=0A= e.style.backgroundPositionX =3D value;=0A= }=0A= else if (name =3D=3D "background-position-y") {=0A= e.style.backgroundPositionY =3D value;=0A= }=0A= else if (name =3D=3D "background-repeat") {=0A= e.style.backgroundRepeat =3D value;=0A= }=0A= else if (name =3D=3D "behavior") {=0A= e.style.behavior =3D value;=0A= }=0A= else if (name =3D=3D "border") {=0A= e.style.border =3D value;=0A= }=0A= else if (name =3D=3D "border-bottom") {=0A= e.style.borderBottom =3D value;=0A= }=0A= else if (name =3D=3D "border-bottom-color") {=0A= e.style.borderBottomColor =3D value;=0A= }=0A= else if (name =3D=3D "border-bottom-style") {=0A= e.style.borderBottomStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-bottom-width") {=0A= e.style.borderBottomWidth =3D value;=0A= }=0A= else if (name =3D=3D "border-collapse") {=0A= e.style.borderCollapse =3D value;=0A= }=0A= else if (name =3D=3D "border-color") {=0A= e.style.borderColor =3D value;=0A= }=0A= else if (name =3D=3D "border-left") {=0A= e.style.borderLeft =3D value;=0A= }=0A= else if (name =3D=3D "border-left-color") {=0A= e.style.borderLeftColor =3D value;=0A= }=0A= else if (name =3D=3D "border-left-style") {=0A= e.style.borderLeftStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-left-width") {=0A= e.style.borderLeftWidth =3D value;=0A= }=0A= else if (name =3D=3D "border-right") {=0A= e.style.borderRight =3D value;=0A= }=0A= else if (name =3D=3D "border-right-color") {=0A= e.style.borderRightColor =3D value;=0A= }=0A= else if (name =3D=3D "border-right-style") {=0A= e.style.borderRightStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-right-width") {=0A= e.style.borderRightWidth =3D value;=0A= }=0A= else if (name =3D=3D "border-style") {=0A= e.style.borderStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-top") {=0A= e.style.borderTop =3D value;=0A= }=0A= else if (name =3D=3D "border-top-color") {=0A= e.style.borderTopColor =3D value;=0A= }=0A= else if (name =3D=3D "border-top-style") {=0A= e.style.borderTopStyle =3D value;=0A= }=0A= else if (name =3D=3D "border-top-width") {=0A= e.style.borderTopWidth =3D value;=0A= }=0A= else if (name =3D=3D "border-width") {=0A= e.style.borderWidth =3D value;=0A= }=0A= else if (name =3D=3D "bottom") {=0A= e.style.bottom =3D value;=0A= }=0A= else if (name =3D=3D "clear") {=0A= e.style.clear =3D value;=0A= }=0A= else if (name =3D=3D "clip") {=0A= e.style.clip =3D value;=0A= }=0A= else if (name =3D=3D "color") {=0A= e.style.color =3D value;=0A= }=0A= else if (name =3D=3D "cssText") {=0A= e.style.Sets =3D value;=0A= }=0A= else if (name =3D=3D "cursor") {=0A= e.style.cursor =3D value;=0A= }=0A= else if (name =3D=3D "direction") {=0A= e.style.direction =3D value;=0A= }=0A= else if (name =3D=3D "display") {=0A= e.style.display =3D value;=0A= }=0A= else if (name =3D=3D "font") {=0A= e.style.font =3D value;=0A= }=0A= else if (name =3D=3D "font-family") {=0A= e.style.fontFamily =3D value;=0A= }=0A= else if (name =3D=3D "font-size") {=0A= e.style.fontSize =3D value;=0A= }=0A= else if (name =3D=3D "font-style") {=0A= e.style.fontStyle =3D value;=0A= }=0A= else if (name =3D=3D "font-variant") {=0A= e.style.fontVariant =3D value;=0A= }=0A= else if (name =3D=3D "font-weight") {=0A= e.style.fontWeight =3D value;=0A= }=0A= else if (name =3D=3D "height") {=0A= e.style.height =3D value;=0A= }=0A= else if (name =3D=3D "ime-mode") {=0A= e.style.imeMode =3D value;=0A= }=0A= else if (name =3D=3D "layout-flow") {=0A= e.style.layoutFlow =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid") {=0A= e.style.layoutGrid =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid-char") {=0A= e.style.layoutGridChar =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid-line") {=0A= e.style.layoutGridLine =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid-mode") {=0A= e.style.layoutGridMode =3D value;=0A= }=0A= else if (name =3D=3D "layout-grid-type") {=0A= e.style.layoutGridType =3D value;=0A= }=0A= else if (name =3D=3D "left") {=0A= e.style.left =3D value;=0A= }=0A= else if (name =3D=3D "letter-spacing") {=0A= e.style.letterSpacing =3D value;=0A= }=0A= else if (name =3D=3D "line-break") {=0A= e.style.lineBreak =3D value;=0A= }=0A= else if (name =3D=3D "line-height") {=0A= e.style.lineHeight =3D value;=0A= }=0A= else if (name =3D=3D "list-style") {=0A= e.style.listStyle =3D value;=0A= }=0A= else if (name =3D=3D "list-style-image") {=0A= e.style.listStyleImage =3D value;=0A= }=0A= else if (name =3D=3D "list-style-position") {=0A= e.style.listStylePosition =3D value;=0A= }=0A= else if (name =3D=3D "list-style-type") {=0A= e.style.listStyleType =3D value;=0A= }=0A= else if (name =3D=3D "margin") {=0A= e.style.margin =3D value;=0A= }=0A= else if (name =3D=3D "margin-bottom") {=0A= e.style.marginBottom =3D value;=0A= }=0A= else if (name =3D=3D "margin-left") {=0A= e.style.marginLeft =3D value;=0A= }=0A= else if (name =3D=3D "margin-right") {=0A= e.style.marginRight =3D value;=0A= }=0A= else if (name =3D=3D "margin-top") {=0A= e.style.marginTop =3D value;=0A= }=0A= else if (name =3D=3D "min-height") {=0A= e.style.minHeight =3D value;=0A= }=0A= else if (name =3D=3D "overflow") {=0A= e.style.overflow =3D value;=0A= }=0A= else if (name =3D=3D "overflow-x") {=0A= e.style.overflowX =3D value;=0A= }=0A= else if (name =3D=3D "overflow-y") {=0A= e.style.overflowY =3D value;=0A= }=0A= else if (name =3D=3D "padding") {=0A= e.style.padding =3D value;=0A= }=0A= else if (name =3D=3D "padding-bottom") {=0A= e.style.paddingBottom =3D value;=0A= }=0A= else if (name =3D=3D "padding-left") {=0A= e.style.paddingLeft =3D value;=0A= }=0A= else if (name =3D=3D "padding-right") {=0A= e.style.paddingRight =3D value;=0A= }=0A= else if (name =3D=3D "padding-top") {=0A= e.style.paddingTop =3D value;=0A= }=0A= else if (name =3D=3D "page-break-after") {=0A= e.style.pageBreakAfter =3D value;=0A= }=0A= else if (name =3D=3D "page-break-before") {=0A= e.style.pageBreakBefore =3D value;=0A= }=0A= else if (name =3D=3D "pixelBottom") {=0A= e.style.pixelBottom =3D value;=0A= }=0A= else if (name =3D=3D "pixelHeight") {=0A= e.style.pixelHeight =3D value;=0A= }=0A= else if (name =3D=3D "pixelLeft") {=0A= e.style.pixelLeft =3D value;=0A= }=0A= else if (name =3D=3D "pixelRight") {=0A= e.style.pixelRight =3D value;=0A= }=0A= else if (name =3D=3D "pixelTop") {=0A= e.style.pixelTop =3D value;=0A= }=0A= else if (name =3D=3D "pixelWidth") {=0A= e.style.pixelWidth =3D value;=0A= }=0A= else if (name =3D=3D "posBottom") {=0A= e.style.posBottom =3D value;=0A= }=0A= else if (name =3D=3D "posHeight") {=0A= e.style.posHeight =3D value;=0A= }=0A= else if (name =3D=3D "position") {=0A= e.style.position =3D value;=0A= }=0A= else if (name =3D=3D "posLeft") {=0A= e.style.posLeft =3D value;=0A= }=0A= else if (name =3D=3D "posRight") {=0A= e.style.posRight =3D value;=0A= }=0A= else if (name =3D=3D "posTop") {=0A= 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quoted-printable Content-Location: http://jnm.snmjournals.org/javascript/entrez/callback.js /************************************************************************= *****=0A= * javascript/entrez/callback.js=0A= *=0A= * Entrez Linking callback to populate content box.=0A= *=0A= * Copyright 2006 Board of Trustees of the Leland Stanford Junior = University.=0A= = *************************************************************************= ***/=0A= =0A= /*=0A= * Execute callback to fill content box with Entrez Linking information.=0A= */=0A= function entrez_callback(pmid, callback_url) {=0A= /*=0A= * MSIE 5.5 and below have issues with the JavaScript=0A= * used for Entrez Linking. 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