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Advances in Brief
Enhanced Antitumor Activity of Anti-epidermal Growth Factor Receptor Monoclonal Antibody IMC-C225 in Combination with Irinotecan (CPT-11) against Human Colorectal Tumor Xenografts
Marie C. Prewett, Andrea T. Hooper, Rajiv Bassi, Lee M. Ellis, Harlan W. Waksal and Daniel J. Hicklin
Marie C. Prewett
Andrea T. Hooper
Rajiv Bassi
Lee M. Ellis
Harlan W. Waksal
Daniel J. Hicklin
DOI: Published May 2002
Colon carcinomas frequently express the epidermal growth factor receptor (EGFR), and this expression correlates with more aggressive disease and poor prognosis. Previous studies have shown that EGFR blockade by monoclonal antibody IMC-C225 can inhibit the growth of human colon carcinoma tumor cells in vitro and xenografts of these tumors in athymic mice. In this report, we have studied the in vivo activity of IMC-C225 combined with the topoisomerase I inhibitor irinotecan (CPT-11) using two models of human colorectal carcinoma in nude mice. IMC-C225 was tested at a dose of 1 or 0.5 mg administered q3d. CPT-11 was administered at a dose of 100 mg/kg/week or a maximum tolerated dose of 150 mg/kg/week. Treatment with the combination of IMC-C225 (1 and 0.5 mg) and CPT-11 (100 mg/kg) significantly inhibited the growth of established DLD-1 and HT-29 tumors compared with either CPT-11 or IMC-C225 monotherapy (P < 0.05). Combination therapy with IMC-C225 (1 mg) and the MTD of CPT-11 (150 mg/kg) resulted in a regression rate of 100 and 60% of established DLD-1 and HT-29 tumors, respectively. In a refractory tumor model, combined treatment with IMC-C225 and CPT-11 significantly inhibited the growth of CPT-11 refractory DLD-1 and HT-29 tumors, whereas either agent alone did not control tumor growth. Histological examination of treated tumors showed extensive tumor necrosis, decreased tumor cell proliferation, increased tumor cell apoptosis, and a marked decrease in tumor vasculature. These results suggest that EGFR blockade by IMC-C225 combined with topoisomerase I inhibitors may be an effective therapy against chemorefractory colorectal carcinoma tumors.
Colorectal cancer is the third most common cancer in men and women in the United States with an estimated 130,200 new cases diagnosed in 2000, including 93,800 patients with colon cancer and 36,400 with rectal cancer (1) . Metastatic colorectal cancer is the second-leading cause of death from cancer in North America, largely because of the poor clinical response of colorectal tumors to conventional chemotherapeutics. The topoisomerase I inhibitor irinotecan (CPT-11) is a water soluble, semisynthetic derivative of camptothecin that has shown activity against a number of different tumor types in preclinical models and in clinical trials of patients with various human cancers (2) . The active metabolite of CPT-11, SN-38, stabilizes a ternary complex between the nuclear enzyme topoisomerase I and double-stranded DNA, which results in replication fork arrest and chromatid breaks (3) . CPT-11 is active clinically in the second-line treatment of patients with colorectal cancer failing or refractory to first-line 5-FU2 and leucovorin therapy (4) . More recently, CPT-11 administered as a three drug regimen along with 5-FU and leucovorin has been shown to prolong survival in patients with colorectal cancer that is refractory to treatment with 5-FU/leucovorin (5) . Despite clinical improvements attributed to the addition of CPT-11 therapy for metastatic colorectal cancer, nearly all patients will eventually become refractory to CPT-11. Thus, new treatment options are needed to improve survival in patients with CPT-11 refractory colorectal cancer.
The EGFR and its ligands EGF and TGF-α play an important role in the growth and survival of human colorectal tumors (reviewed in Refs. 6 and 7 ). EGF and/or TGFα mRNA and protein are found in a majority of human colorectal tumors and cell lines, and coexpression of these ligands and EGFR has been reported, suggesting the potential existence of an autocrine loop. In addition, the expression of EGFR in colorectal cancer correlates with more aggressive disease and poor prognosis. Hence, blockade of the EGFR receptor may be a useful strategy for treatment of adenocarcinoma of the colon. IMC-C225 binds the EGFR with high affinity, competes for ligand binding, and down-regulates receptor expression on the cell surface (reviewed in Ref. 8 ). Several studies have shown that IMC-C225 is capable of inhibiting growth of EGFR-expressing tumor cells in vitro, and treatment with IMC-C225 results in marked inhibition of tumor growth in nude mice bearing xenografts of human cancer cell lines (8, 9, 10) . Moreover, treatment with IMC-C225 in combination with chemotherapeutic drugs or radiotherapy is effective in eradicating well-established tumors in nude mice (8, 9, 10, 11) . IMC-C225 is currently in Phases II and III clinical testing in a number of EGFR+ malignancies (9) . The objective of this study was to determine the potential therapeutic utility of the MAb IMC-C225 when combined with CPT-11 therapy to inhibit growth of human colon carcinoma tumor xenografts in a nude mouse model.
The colon carcinoma cell lines DLD-1 and HT-29 were obtained from the American Type Culture Collection (Manassas, VA) and maintained in DMEM (DLD-1) or McCoy's 5A (HT-29) media (Invitrogen, Carlsbad, CA) supplemented with 10% fetal bovine serum (HyClone, Lenexa, KY) and 2 mm GlutaMAX (Invitrogen). Cells were cultured at 37°C in a 5% CO2 atmosphere and passaged routinely by Trypsin-EDTA (Invitrogen) treatment.
Anti-EGFR MAb IMC-C225
Clinical grade anti-EGFR MAb IMC-C225 (ERBITUX) was produced by the ImClone Systems, Inc. manufacturing facility (Somerville, NJ).
Subconfluent cultures of colon carcinoma cell lines, grown in 100-mm2 plates, were washed in ice-cold HBSS and detached by incubating the monolayers in HBSS with EDTA. Aliquots of 106 cells were incubated for 1 h on ice with primary antibody (IMC-C225 or control IgG), diluted in PBS with 1% BSA and 0.02% sodium azide (flow buffer). Cells were washed twice with flow buffer and then incubated for 30 min on ice with FITC-labeled goat antihuman IgG Fc-specific secondary antibody (KPL, Gaithersburg, MD) diluted in flow buffer. Cells were washed as above and analyzed for FITC emission intensity on an Epics XL (Beckman-Coulter, Hialeah, FL) flow cytometer. Dead cells and debris were eliminated from the analysis on the basis of forward and sideways light scatter.
Phosphorylation Assay
Phosphorylation assays were done by seeding DLD-1 or HT-29 cells at 5 × 106/150-mm2 tissue culture plate in media containing 0.5% fetal bovine serum in 5 μg/ml IMC-C225 or normal human IgG (Jackson Immunoresearch, West Grove, PA). After culturing for 24 h, cells were then stimulated with 5 ng/ml EGF or 5 ng/ml TGF-α (Sigma Chemical Co., St. Louis, MO) for 20 min at 37°C. Monolayers were than washed with ice-cold PBS containing 1 mm sodium orthovanadate. Cells were lysed, immunoprecipitated with IMC-C225, and subjected to Western blot analysis. The phosphorylation patterns were determined by probing the blots with a horseradish peroxidase-conjugated antiphosphotyrosine MAb (UBI) followed by detection using the Lumi-GLO method (KPL). Loading equivalence of EGFR in each lane was verified by stripping and reprobing blots with a polyclonal anti-EGFR antibody (Calbiochem, San Diego, CA).
Female athymic (nu/nu) mice (5–6 weeks old) were purchased from Harlan Sprague Dawley, Inc. (Indianapolis, IN). Mice were housed under pathogen-free conditions in microisolator cages with laboratory chow and water available ad libitum. All experiments and procedures were performed in accordance with the United States Department of Agriculture, Department of Health and Human Services, and NIH policies regarding the humane care and use of laboratory animals.
Treatment of s.c. Colorectal Carcinoma Xenografts
Subcutaneous colon carcinoma tumors were established by injecting 5 × 106 DLD-1 or HT-29 cells mixed in Matrigel (Collaborative Research Biochemicals, Bedford, MA) s.c. into the left flank of athymic mice. Three separate study designs were used to examine the activity of IMC-C225, irinotecan (CPT-11; Pharmacia & Upjohn, Kalamazoo, MI), and combination therapy.
Study Design I.
Tumors were allowed to reach 150–300 mm3 in size, and then mice were randomized into groups of five animals each. Mice were treated with saline q3d, IMC-C225 at 1 mg/dose (q3d), IMC-C225 at 0.5 mg/dose (q3d), CPT-11 at 100 mg/kg (q7d), combination treatment with IMC-C225 at 1 mg/dose (q3d) + CPT-11 at 100 mg/kg (q7d), and IMC-C225 at 0.5 mg/dose (q3d) + CPT-11 at 100 mg/kg (q7d). Treatment of animals was continued for the duration of the study.
Study Design II.
Tumors were allowed to reach 150–300 mm3 in size, and then mice were randomized into groups of 10 animals each. Mice were treated with saline q3d, IMC-C225 at 1 mg/dose (q3d), CPT-11 at 150 mg/kg (q7d), or combination treatment with IMC-C225 at 1 mg/dose (q3d) + CPT-11 at 150 mg/kg (q7d). Treatment of animals was continued for the duration of the study.
Study Design III.
Tumors were allowed to reach 150 mm3 in size, and then mice were treated with two cycles of CPT-11 (100 mg/kg; days 0 and 7). After two cycles of CPT-11 treatment (day 12), those CPT-11-treated mice with tumor volumes > 2-fold the volume on day 0 ("refractory" mice) were randomized into three groups of seven to eight mice per group. The frequency of CPT-11 refractory mice was 50–60% of the total CPT-11-treated group. CPT-11 refractory mice were treated with IMC-C225 at 1 mg/dose (q3d), CPT-11 at 100 mg/kg (q7d), or combination treatment with IMC-C225 at 1 mg/dose (q3d) + CPT-11 at 100 mg/kg (q7d). Treatment of animals was continued for the duration of the study.
The MTD for CPT-11 therapy was defined as the maximum dose that did not result in drug-related morbidity and/or weight loss of >20% of body weight at the beginning of therapy. Mice were monitored daily for signs of CPT-11-related toxicity (body weight, diarrhea, and lethality).
All drug treatments were administered by i.p. injection. Tumors were measured twice each week with calipers, and tumor volumes were calculated by the formula [π/6 (w1 × w2 × w2)], where "w1" represents the largest tumor diameter, and "w2" represents the smallest tumor diameter.
Tumors were harvested from animals at multiple time points after treatment and snap frozen in liquid nitrogen for anti-CD31 immunohistochemistry and TUNEL staining. For anti-CD31 immunohistochemistry, 6-μm cryostat sections were fixed at room temperature in acetone for 5 min and were postfixed in 10% neutral buffered formalin for 10 s before staining. Endogenous peroxidase activity was blocked by a 30-min incubation with 0.3% H2O2 in H2O. After blocking with 5% BSA, 10% normal goat serum + 0.02% Tween 20, FITC anti-CD31 (clone Mec13.3, BD PharMingen, Franklin Lakes, NJ), and FITC rat IgG isotype control (PharMingen) were diluted to a final concentration of 1 μg/ml in blocking buffer and were incubated with the tissue sections for 60 min at room temperature. Slides were then washed in PBS to remove excess antibody. Slides were incubated with 1 μg/ml horseradish peroxidase-conjugated anti-FITC (Roche Molecular Biochemicals, Indianapolis, IN) for 30 min at room temperature and were washed subsequently in PBS. Immunostaining was developed for 4 min using a liquid DAB substrate kit (Zymed, San Francisco, CA) per manufacturer's instructions. The reactions were stopped by successive washes in tap and distilled water and were lightly counterstained with Mayer's hematoxylin. Sections were coverslipped using a permanent mounting medium. TUNEL staining was performed using the Apoptosis Detection System, Fluorescein (Promega, Madison, WI). Briefly, 6-μm cryostat sections were fixed in 4% paraformaldehyde for 10 min at room temperature and rinsed in PBS with 0.1% Triton X-100. Sections were then incubated in Equilibration Buffer for 5 min at room temperature followed by incubation in TUNEL Mix, prepared according to kit instructions, for 1 h at 37°C. After successive washes in PBS, sections were coverslipped using an antifade reagent.
For H&E and Ki-67 staining, resected tumors were fixed in zinc-buffered formalin (Shandon Lipshaw, Pittsburgh, PA) overnight at 4°C. After paraffin embedding and sectioning at 6 μm, formalin-fixed sections were stained with Mayer's H&E (Richard Allen, Kalamazoo, MI). For Ki-67 immunohistochemistry, slides were heated in a water bath at 95°C-99°C in Target Retrieval Solution (DAKO, Carpinteria, CA) for 20 min, followed by a 20-min cool down period at room temperature. After heat retrieval, sections were rinsed well in PBS and stained with rabbit antihuman Ki-67 antigen (DAKO N-series, ready to use) per manufacturer's instructions and were lightly counterstained with Mayer's hematoxylin.
The Proliferation Index was determined by Ki-67 immunostaining and calculating the number of DAB-positive pixels per total number of nuclear pixels (hematoxylin-positive pixels plus DAB-positive pixels × 100) in five fields at ×200. The Apoptosis Index, determined by TUNEL staining, is calculated from the number of TUNEL-positive pixels per total number of Hoechst-positive pixels × 100 in five fields at ×200. The Apoptosis:Proliferation Ratio equals the Apoptosis Index/Proliferation Index × 1000.
Tumor volumes and histological quantitations were analyzed using Student's t test. Antitumor activity of combination therapy with IMC-C225 and CPT-11 in vivo was analyzed by the fractional product method as described (12) . Analyses were computed using the SigmaStat statistical package (v. 2.03; Jandel Scientific, San Rafael, CA).
The in vivo effects of IMC-C225 and CPT-11 therapy alone or in combination were tested in a DLD-1 and HT-29 s.c. xenograft model in athymic nude mice. DLD-1 and HT-29 cells express moderate levels of EGFR, and IMC-C225 inhibits both EGF and TGF-α-stimulated EGFR tyrosine phosphorylation in both cell lines (Fig. 1, A and B)⇓ . Three different therapy designs were tested in the DLD-1 and HT-29 tumor models. In therapy schedule I, mice with well-established (∼200 mm3) DLD-1 or HT-29 tumors were treated by i.p. administration of two different doses of IMC-C225 (1 or 0.5 mg; q3d) with or without CPT-11 (100 mg/kg; q7d). A total of 16 cycles of IMC-C225 and 7 cycles of CPT-11 was administered. Treatment with IMC-C225, CPT-11, or the combination was well tolerated for the duration of therapy. IMC-C225 treatment alone at 1- or 0.5-mg doses did not significantly inhibit growth of DLD-1 or HT-29 tumors (P > 0.05; Fig. 2, A and B⇓ ). Although CPT-11 treatment alone (100 mg/kg/week) was active against DLD-1 (P < 0.02) tumors, the treatment did not significantly inhibit growth of HT-29 tumors (P > 0.05). In contrast, enhanced antitumor activity using combination therapy with IMC-C225 and CPT-11 was observed in both DLD-1 and HT-29 tumors compared with mice treated with control, IMC-C225, or CPT-11 alone.
A, EGFR expression on DLD-1 and HT-29 colon carcinoma cells. Flow cytometry analysis histograms of IMC-C225 binding (black) or a control human antibody (gray). B, phosphorylation assay showing IMC-C225 inhibition of EGFR phosphorylation. DLD-1 or HT-29 cultures were incubated with IMC-C225 or normal human IgG followed by stimulation with exogenous EGF or TGF-α. Western blotting with antiphosphotyrosine antibody (top panels). Blots were stripped and reprobed with anti-EGFR antibody (bottom panels).
Growth inhibition of DLD-1 and HT-29 colorectal tumor xenografts in nude mice. A and B, dose-dependent effects are shown for treatment of established DLD-1 (A) and HT-29 (B) tumors treated with saline (○), IMC-C225 at 1 mg (•) or 0.5 mg (▵) per dose/q3d, CPT-11 at 100 mg/kg/week (□), CPT-11 (100 mg/kg/week) plus IMC-C225 at 1 mg/dose/q3d (▪), and CPT-11 (100 mg/kg/week) plus IMC-C225 at 0.5 mg/dose/q3d (▴). C and D, combination therapy on established DLD-1 (C) and HT-29 (D) tumors treated with saline (○), IMC-C225 at 1 mg (•), CPT-11 at 150 mg/kg/week (□), and CPT-11 (150 mg/kg/week) plus IMC-C225 at 1 mg/dose/q3d (▪). Bars, ±SE.
In the second study design, we attempted to achieve a maximal therapeutic effect by increasing the dose of CPT-11 to the MTD. Pilot studies determined that the MTD of CPT-11 in mice was 150 mg/kg when given q7d (data not shown). The MTD of CPT-11 was then used in combination therapy with IMC-C225 in the DLD-1 and HT-29 xenograft models (Fig. 2, C and D)⇓ . A total of 17 cycles of IMC-C225 and 7 cycles of CPT-11 was administered. Combination therapy with 1 mg of IMC-C225 and 150 mg/kg CPT-11 resulted in a significant inhibition of tumor growth compared with single-agent IMC-C225 (P < 0.002) or compared with single-agent CPT-11 (P < 0.03). In the DLD-1 model, complete inhibition of tumor growth was observed in the majority of animals. In addition, regression of established DLD-1 tumors was observed in 60% of animals treated with the combination of IMC-C225 and CPT-11. The activity of IMC-C225 and CPT-11 treatment was even more pronounced in the HT-29 model. Regression of established HT-29 tumors was observed in 100% of the animals treated with IMC-C225 and CPT-11 combination therapy. The indices for combination therapy are >1, indicating an enhanced interaction between IMC-C225 and CPT-11. Table 1⇓ summarizes the analysis of the enhanced combination effect at different time points during therapy.
Combination therapy with IMC-C225 and CPT-11
The results from studies described above demonstrated that DLD-1 or HT-29 tumors are poorly responsive to IMC-C225 or CPT-11 therapy alone. To further examine the effect of IMC-C225/CPT-11 combination therapy against CPT-11 refractory colorectal tumors, a refractory model was designed and tested. Animals with established DLD-1 or HT-29 tumors were treated with two cycles of CPT-11 (100 mg/kg; days 0 and 7). At day 12, tumor volumes were measured, and animals with tumors that had increased >2 × the original tumor volume at the start of CPT-11 therapy were considered CPT-11 refractory. These mice were selected and randomized into treatment groups receiving continued CPT-11 (100 mg/kg; q7d), switched to IMC-C225 therapy (1 mg; q3d) or combination therapy on the same schedule. DLD-1 or HT-29 tumors in animals that continued to receive CPT-11 grew at similar kinetics before randomization, i.e., these tumors did not respond to additional CPT-11 therapy (Fig. 3)⇓ . IMC-C225 therapy alone did not have a significant antitumor effect on CPT-11 refractory DLD-1 or HT-29 tumors and grew at similar kinetics to those tumors treated with CPT-11 (P > 0.05 compared with CPT-11-treated mice). In contrast, combination therapy with IMC-C225 and CPT-11 significantly inhibited the growth of CPT-11 refractory DLD-1 or HT-29 tumors (P < 0.01 and <0.01, respectively) with enhanced antitumor activity compared with either agent alone (Table 2)⇓ .
Growth inhibition of CPT-11 refractory colorectal tumor xenografts in nude mice. Mice with established DLD-1 (A) or HT-29 (B) tumors were treated with two cycles of CPT-11 therapy (100 mg/kg) on days 0 and 7. Mice with tumors that did not respond to CPT-11 therapy (>2 × initial tumor volume at day 12; shown as dotted vertical line) were selected, randomized, and then treated with IMC-C225 at 1 mg/dose/q3d (•), continued CPT-11 at 100 mg/kg/week (□), or combination therapy (▪). Bars, ±SE.
Response of CPT-11 refractory xenografts to IMC-C225 and CPT-11
Histological examination of HT-29 xenografts at day 47 after treatment showed marked differences in tumors from animals administered the combined IMC-C225/CPT-11 therapy compared with tumors from animals receiving control or single agent therapy. Extensive tumor necrosis was observed in DLD-1 and HT-29 tumors treated with a combination of IMC-C225 and CPT-11 (Fig. 4A)⇓ . A reduction in tumor cell proliferation (Ki-67 staining), a reduction in tumor vasculature (CD31 staining), and an increase in tumor apoptosis (TUNEL staining) was observed in tumors treated with IMC-C225 and CPT-11. Quantitative analysis of the tumor cell proliferation, apoptosis, and vessel staining showed a significant difference between combination therapy versus control and between combination therapy versus single-agent therapy (P < 0.001 for all comparisons; Fig. 4B⇓ ). In HT-29 tumors, combination treatment resulted in a ≥1.4-fold decrease in tumor cell proliferation, a ≥2.9-fold increase in tumor cell apoptosis, and a ≥4.3-fold decrease in vessel staining. The Apoptosis:Proliferation Ratio was increased ≥4.5-fold in IMC-C225/CPT-11-treated tumors compared with either agent alone. Similar findings were observed in DLD-1 tumors. These findings show that the antitumor activity of combined treatment with IMC-C225 and CPT-11 is a result of increased tumor cell apoptosis and necrosis compared with treatment with either agent alone.
A, histological examination of HT-29 tumor xenografts (day 47) stained with H&E, anti-CD31 vessel staining (brown), anti-Ki-67 nuclear antigen (black), and apoptosis by FITC-labeled TUNEL assay (green). (magnification: ×200). A higher magnification of combination therapy-treated HT-29 tumors stained with H&E is shown (inset) to illustrate necrosis in these tumors (×400). B, quantitation of CD31 vessel staining, Ki-67 Proliferation Index, Apoptosis Index, and Apoptosis:Proliferation Ratio. Columns, mean number positive pixels in 10 fields. Bars, ±SD. Tumors from mice treated with vehicle (white), IMC-C225 (diagonal hatched), CPT-11 (horizontal hatched), or IMC-C225 plus CPT-11 (cross-hatched).
EGFR and its ligands EGF and TGF-α are important factors in the normal physiology of the human gastrointestinal tract (for review, see Ref. 13 ). EGF is a potent epithelial mitogen that is produced in the Paneth cells of the small intestine and the Brunner's glands of the duodenum (14) . EGF stimulates intestinal crypt stem cell proliferation in both the small and large intestine and is an important survival factor for the intestinal mucosa (14 , 15) . EGFR and TGF-α expression can be detected in normal epithelial mucosa throughout the gastrointestinal tract (16) . The EGFR and its ligands also play an important role in the growth and survival of human colorectal tumors (6 , 7) . EGFR is expressed at varying levels in human colorectal tumors and colorectal carcinoma cell lines. Moreover, coexpression of EGF and/or TGFα and EGFR has been reported in several human colon tumor cell lines, suggesting the potential existence of an autocrine stimulatory loop (17, 18, 19) . In addition, studies have demonstrated that the EGFR expression level is related to the progression of the colon cancer disease, as well as survival after tumor resection (20 , 21) . The relationship between EGFR expression and the hepatic metastasis of colon adenocarcinoma has been reported in a limited number of studies (22) . These studies suggest that expression of EGFR in colon carcinoma correlates with the ability to produce liver metastases. Hence, the role of the EGFR signaling pathway in colorectal cancer has led to considerable interest in the potential therapeutic utility of agents that block EGFR function.
Studies herein were performed to determine the activity of the anti-EGFR MAb IMC-C225 in combination with the topoisomerase I inhibitor CPT-11 in colorectal tumor models and to provide a rationale for clinical development of this combination. We hypothesized that combining IMC-C225 with CPT-11 would enhance the antitumor activity of these drugs against tumors that are poorly responsive to either drug alone. Our findings show that functional inhibition of EGFR with IMC-C225 combined with CPT-11 therapy is highly effective in two different colorectal tumor models. Importantly, HT-29 and DLD-1 colorectal tumor xenografts were found to be poorly sensitive to CPT-11 or IMC-C225 monotherapy. However, when IMC-C225 treatment was added to CPT-11 therapy, enhanced tumor growth inhibition was observed. To further evaluate the enhanced effects of combining IMC-C225 with CPT-11, we studied the combination of these two agents in a more rigorous CPT-11 refractory model. In these experiments, mice with established HT-29 or DLD-1 tumors that did not respond to CPT-11 therapy were selected and subsequently treated with CPT-11, IMC-C225, or the combination. Remarkably, IMC-C225 treatment was able to reverse the CPT-11 refractory nature of both HT-29 and DLD-1 tumors. These results are consistent with a number of studies demonstrating the same principle; namely, combined therapy with IMC-C225 and antineoplastic drugs results in a marked potentiation of antitumor activity compared with monotherapy. IMC-C225 or its parent murine antibody 225 has been shown to augment the antitumor activity of several anticancer agents, including cisplatin, doxorubicin, 5-FU, gemcitabine, and paclitaxel in preclinical models (23, 24, 25, 26, 27) . Similar enhancement of activity has been observed in studies where IMC-C225 is used in combination with irradiation (11 , 28 , 29) . Importantly, the combination of IMC-C225 with cytotoxic drugs/radiation often results in tumor regression and, in many cases, eradication of established tumors. It should be noted that overexpression of EGFR by tumor cells is not required for the enhanced activity observed when combining IMC-C225 with chemotherapy and particular in these studies when combining IMC-C225 with CPT-11. The two colorectal tumor lines used in our studies express moderate levels of EGFR (∼1 × 105) and respond to ligand stimulation, and receptor activation can be inhibited by treatment with IMC-C225. These findings suggest that the biology of the EGFR pathway in tumor cells, rather than the overall receptor level, is important for the activity of IMC-C225 when combined with chemotherapeutic agents. The effectiveness of combining IMC-C225 and chemotherapy or radiation therapy observed in preclinical studies has been extended to clinical trials in patients with squamous cell carcinoma of the head and neck (30 , 31) .
The activity of IMC-C225 or its parental murine antibody 225 has been demonstrated previously in a number of in vitro and in vivo preclinical models of human colon adenocarcinoma (32, 33, 34) . In these studies, treatment with IMC-C225 inhibited the growth of colorectal tumor cells in vitro and growth of s.c. tumor xenografts. Combination treatment with IMC-C225 and other antineoplastic agents or radiation has been shown to enhance the antitumor effect against established colorectal tumor xenografts (34, 35, 36) . Ciardiello et al. (35) tested IMC-C225 in combination with the topoisomerase I inhibitor topotecan in the human GEO colon adenocarcinoma xenograft model. Treatment of mice bearing established GEO colon tumors with either IMC-C225 or topotecan alone was ineffective at controlling the growth of GEO tumors. However, combined treatment resulted in a significant suppression of tumor growth and almost complete tumor regression and a significant survival advantage compared with animals receiving monotherapy. These results are consistent with those obtained in the present study with CPT-11. Interestingly, Ciardiello et al. (35) found that sequential treatment with topotecan followed by IMC-C225 therapy resulted in the most effective antitumor response. In our studies, CPT-11 and IMC-C225 was administered to mice simultaneously during the initial treatment cycle followed by weekly CPT-11 treatment with sustained serum levels of IMC-C225 administered on a q3d schedule. We found no dependency for sequential treatment with CPT-11 followed by IMC-C225. Moreover, we have found optimal therapeutic effects in animal models when CPT-11 and IMC-C225 are administered together rather than sequentially.3 The discrepancy between our results and that of Ciardiello et al. (35) are unclear. These differences may be related to drug treatment schedule, pharmacodynamics of CPT-11 versus topotecan, mechanisms of interaction between IMC-C225 and CPT-11 versus topotecan, or differences in the tumor models.
Histological examination of HT-29 and DLD-1 tumors treated with IMC-C225 and CPT-11 therapy showed marked changes in several markers that can be attributed to the antitumor effect of combination therapy. These comprise a decrease in tumor cell proliferation, an increase in tumor cell apoptosis, and a reduction in tumor vasculature. The reduction in tumor cell proliferation and tumor vasculature and increase in apoptosis led to extensive tumor necrosis in IMC-C225/CPT-11-treated colorectal tumors. The incidence of tumor cell apoptosis suggests that the effects of IMC-C225/CPT-11 treatment on colon carcinoma tumors in vivo were not merely cytostatic but rather affect the growth and survival of these tumors. These findings are consistent with those of previous studies that have demonstrated IMC-C225-induced tumor cell apoptosis in other EGFR+ human tumor models in nude mice (25 , 27 , 37) . The decreased CD31 vessel staining observed in HT-29 and DLD-1 xenografts are also consistent with results of other studies and suggest that antiangiogenic mechanisms may be associated with IMC-C225 treatment (25 , 37 , 38) . However, given the dramatic antitumor effect of combination therapy observed in these studies, it is difficult to determine to what extent generalized tumor necrosis contributed to the reduction of microvessels in treated tumors. We speculate that reduced vasculature in HT-29 and DLD-1 tumors treated with IMC-C225 and CPT-11 is because of both generalized tumor necrosis and a direct effect on neoangiogenesis.
A number of mechanisms have been identified that characterize the effect of IMC-C225 on the growth and survival of EGFR+ tumors in preclinical models (reviewed in Ref. 8 ). The direct mechanism responsible for IMC-C225's activity is inhibition of EGFR tyrosine kinase activation. The outcome of this blockade is reflected in the disruption of any number of processes regulated by the EGFR pathway in a given tumor cell. These processes include: (a) regulation of cell cycle progression; (b) cell survival pathways; (c) tumor cell invasion; and (d) angiogenesis. Several mechanisms appear to contribute to the enhanced cytotoxic response observed with combinations of IMC-C225 and chemo/radiation. Genotoxic damage induced by chemotherapy or radiation leads to induction of arrest at the G1-S cell cycle checkpoint or activation of apoptosis; both are regulated by the EGFR signaling pathway. Disruption of EGFR-mediated survival signals and an increase in programmed cell death appear to be major mechanisms whereby IMC-C225 synergizes with cytotoxic agents (8 , 11) . Changes in the expression or activation of a number of molecules after IMC-C225 treatment contribute to this response; these include p27 (Kip1), cdk-2, Rb, Bcl-2, and Bax. Studies of sublethal DNA damage repair and potentially lethal damage repair analyses in cultured tumor cells have demonstrated a strong inhibitory effect of IMC-C225 on postradiation DNA damage repair (11) . These studies have also shown that exposure of tumor cells to IMC-C225 induces a redistribution of DNA-dependent protein kinase from the nucleus to the cytosol and reduction in proliferating cell nuclear antigen expression, which is known to play a role in DNA repair (39) . Hence, IMC-C225 inhibits both survival pathways and DNA repair mechanisms and, thus, enhances the apoptotic potential of chemotherapeutic drugs or radiation that induce cellular damage and programmed cell death. Finally, IMC-C225 may also enhance the response to cytotoxic therapy through mechanisms involving reduced angiogenesis and tumor cell invasion. The mechanisms responsible for the enhanced antitumor activity we observed in the present studies have not been fully defined. Our findings from analysis of HT-29 and DLD-1 tumors suggest that the enhanced antitumor effects IMC-C225/CPT-11 therapy are attributable to cell cycle, apoptotic, and possibly angiogenic mechanisms of tumor growth. However, additional tumorigenic regulatory processes, such as DNA repair and unknown mechanisms of CTP-11 resistance, are likely to play a contributing role. Additional studies will be necessary to fully elucidate the mechanisms contributing to the IMC-C225/CPT-11 antitumor response and determine which molecular pathways are involved in this response.
In conclusion, these results demonstrate enhanced antitumor activity of IMC-C225 combined with CPT-11 therapy compared with CPT-11 or IMC-C225 treatment alone. This combination was highly effective against established, CPT-11 refractory colorectal tumors. The precise mechanisms responsible for the combined effects of IMC-C225 and CPT-11 in these colorectal tumor models are still unclear and will require additional studies. Nevertheless, the ability of combination therapy to inhibit growth of chemoresistant colorectal carcinoma tumors suggests that EGFR blockade by IMC-C225 combined with CPT-11 has potential as a promising therapeutic strategy for clinical testing in CPT-11 refractory colorectal cancer.
We thank Dr. Peter Bohlen for critical review of the manuscript and Sarah Michaud for assistance in preparation of the manuscript. We dedicate this work to the memory of our good friend and colleague, Angel Santiago.
↵1 To whom requests for reprints should be addressed, at ImClone Systems, Inc., Department of Immunology, 180 Varick Street, New York, New York 10014. Phone: (212) 645-1405; Fax: (212) 645-2054; E-mail: DanH{at}imclone.com
↵2 The abbreviations used are: 5-FU, 5-fluorouracil; EGFR, epidermal growth factor receptor; CPT-11, 7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxy-camptothecin (irinotecan); EGF, epidermal growth factor; TGF, transforming growth factor; MAb, monoclonal antibody; MTD, maximum tolerated dose; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling; DAB, diaminobenzidine.
↵3 M. C. Prewett, unpublished data.
Received September 26, 2001.
Accepted February 16, 2002.
Greenlee R. T., Murray T., Bolden S., Wingo P. A. Cancer statistics, 2000. CA Cancer J. Clin., 50: 7-33, 2000.
Saijo N. Preclinical and clinical trials of topoisomerase inhibitors. Ann. N. Y. Acad. Sci., 922: 92-99, 2000.
Rivory L. P. Irinotecan (CPT-11): a brief overview. Clin. Exp. Pharmacol. Physiol., 23: 1000-1004, 1996.
Rothenberg M. L., Cox J. V., DeVore R. F., Hainsworth J. D., Pazdu R., Rivkin S. E., Macdonald J. S., Geyer C.E., Jr., Sandbach J., Wolf D. L., Mohrland J. S., Elfring G. L., Miller L. L., Von Hoff D. D. A multicenter, phase II trial of weekly irinotecan (CPT-11) in patients with previously treated colorectal carcinoma. Cancer (Phila.), 85: 786-795, 1999.
Saltz L. B., Cox J. V., Blanke C., Rosen L. S., Fehrenbacher L., Moore M. J., Maroun J. A., Ackland S. P., Locker P. K., Pirotta N., Elfring G. L., Miller L. L. Irinotecan plus fluorouracil and leucovorin for metastatic colorectal cancer. Irinotecan study group. N. Engl. J. Med., 343: 905-914, 2000.
Salomon D. S., Bradt R., Ciardiello F., Normanno N. Epidermal growth factor-related peptides and their receptors in human malignancies. Crit. Rev. Oncol. Hematol., 19: 183-232, 1995.
Normanno N., De Luca A., Salomon D. S., Ciardiello F. Epidermal growth factor-related peptides as targets for experimental therapy of human colon carcinoma. Cancer Detect. Prev., 22: 62-67, 1998.
Mendelsohn J. Blockade of receptors for growth factors: an anticancer therapy. Clin. Cancer Res., 6: 747-753, 2000.
Mendelsohn J. The epidermal growth factor receptor as a target for cancer therapy. Endocr. Relat. Cancer, 8: 3-9, 2001.
Waksal H. W. Role of an anti-epidermal growth factor receptor in treating cancer. Cancer Metastasis Rev., 18: 427-436, 1999.
Huang S. M., Harari P. M. Modulation of radiation response after epidermal growth factor receptor blockade in squamous cell carcinomas: inhibition of damage repair, cell cycle kinetics and tumor angiogenesis. Clin. Cancer Res., 6: 2166-2174, 2000.
Yokoyama Y., Dhanabal M., Griffioen A. W., Sukhatme V. P., Ramakrishnan S. Synergy between angiostatin and endostatin: inhibition in ovarian cancer growth. Cancer Res., 60: 2190-2196, 2000.
Chailler P., Menard D. Ontogeny of EGF receptors in the human gut. Front. Biosci., 4: D87-D101, 1999.
Reeves J. R., Richards R. C., Cooke T. The effects of intracolonic EGF on mucosal growth and experimental carcinogenesis. Br. J. Cancer, 63: 223-226, 1991.
Wright N. A., Pike C., Elia G. Induction of a novel epidermal growth factor-secreting cell lineage by mucosal ulceration in human gastrointestinal stem cells. Nature (Lond.), 343: 82-85, 1990.
Markowitz S. D., Molkentin K., Gerbic C., Jackson J., Stellato T., Wilson J. K. V. Growth stimulation by coexpression of transforming growth factor-α and epidermal growth factor receptor in normal and adenomatous human colon epithelium. J. Clin. Investig., 86: 356-362, 1990.
Huang S. M., Lin P. F., Fan D., Price J. E., Trujillo J. M., Chakrabarty S. Growth modulation by epidermal growth factor (EGF) in human colonic carcinoma cells: constitutive expression of the human EGF gene. J. Cell. Physiol., 148: 220-227, 1991.
Coffey R. J., McCutchen C. M., Graves-Deal R., Polk W. H., Jr. Transforming growth factors and related peptides in gastrointestinal neoplasia. J. Cell. Biochem. Suppl., 16G: 111-118, 1992.
Howell G. M., Sun L. Z., Ziober B. L., Wu S. P., Brattain M. G. The role of growth regulatory aberrations in progression of human colon carcinoma. Cancer Metastasis Rev., 12: 275-286, 1993.
Mayer A., Takimoto M., Fritz E., Schellander G., Kofler K., Ludwig H. The prognostic significance of proliferating cell nuclear antigen, epidermal growth factor receptor, and mdr gene expression in colorectal cancer. Cancer (Phila.), 71: 2454-2460, 1993.
De Jong K. P., Stellema R., Karrenbeld A., Koudstaal J., Gouw A. S., Sluiter W. J., Peeters P. M., Slooff M. J., De Vries E. G. Clinical relevance of transforming growth factor α, epidermal growth factor receptor, p53, and Ki67 in colorectal liver metastases and corresponding primary tumors. Hepatology, 28: 971-979, 1998.
Radinksy R., Risin, Fan, Dong, Bielenberg, Bucana C. D., Fidler J. Level and function of epidermal growth factor receptor predict the metastatic potential of human colon carcinoma cells. Clin. Cancer Res., 1: 19-31, 1995.
Baselga J., Norton L., Masui H., Pandiella A., Coplan K., Miller W. H., Jr., Mendelsohn J. Antitumor effects of doxorubicin in combination with anti-epidermal growth factor receptor monoclonal antibodies. J. Natl. Cancer Inst. (Bethesda), 85: 1327-1333, 1993.
Prewett M., Rockwell P., Rose C., Goldstein N. I. Anti-tumor and cell cycle responses in KB cells treated with a chimeric anti-EGFR monoclonal antibody in combination with cisplatin. Int. J. Oncol., 9: 217-224, 1996.
Bruns C. J., Harbison M. T., Davis D. W., Portera C. A., Tsan R., McConkey D. J., Evans D. B., Abbruzzese J. L., Hicklin D. J., Radinsky R. Epidermal growth factor receptor blockade with C225 plus gemcitabine results in regression of human pancreatic carcinoma growing orthotopically in nude mice by antiangiogenic mechanisms. Clin. Cancer Res., 6: 1936-1948, 2000.
Overholser J. P., Prewett M. C., Hooper A. T., Waksal H. W., Hicklin D. J. Epidermal growth factor receptor blockade by antibody IMC-C225 inhibits growth of human pancreatic carcinoma xenografts in nude mice. Cancer (Phila.), 89: 74-82, 2000.
Inoue K., Slaton J. W., Perrotte P., Davis D. W., Bruns C. J., Hicklin D. J., McConkey D. J., Sweeney P., Radinsky R., Dinney C. P. Paclitaxel enhances the effects of the anti-epidermal growth factor receptor monoclonal antibody ImClone C225 in mice with metastatic human bladder transitional cell carcinoma. Clin. Cancer Res., 6: 4874-4884, 2000.
Saleh M. N., Raisch K. P., Stackhouse M. A., Grizzle W. E., Bonner J. A., Mayo M. S., Kim H-G., Meredith R. F., Wheeler R. H., Buchsbaum D. J. Combined modality therapy of A431 human epidermoid cancer using anti-EGFr antibody C225 and radiation. Cancer Biother. Radiopharm., 14: 451-463, 1999.
Milas L., Mason K., Hunter N., Petersen S., Yamakawa M., Ang K., Mendelsohn J., Fan Z. In vivo enhancement of tumor radioresponse by C225 anti-epidermal growth factor receptor antibody. Clin. Cancer Res., 6: 701-708, 2000.
Robert F., Ezekiel M. P., Spencer S. A., Meredith R. F., Bonner J. A., Khazaeli M. B., Saleh M. N., Carey D., LoBuglio A. F., Wheeler R. H., Cooper M. R., Waksal H.W. Phase I study of anti-epidermal growth factor receptor antibody cetuximab in combination with radiation therapy in patients with advanced head and neck cancer. J. Clin. Oncol., 19: 3234-3243, 2001.
Shin D. M., Donato N. J., Perez-Soler R., Shin H. J., Wu J. Y., Zhang P., Lawhorn K., Khuri F. R., Glisson B. S., Myers J., Clayman G., Pfister D., Falcey J., Waksal H., Mendelsohn J., Hong W. K. Epidermal growth factor receptor-targeted therapy with C225 and cisplatin in patients with head and neck cancer. Clin. Cancer Res., 7: 1204-1213, 2001.
Karnes W. E., Jr., Walsh J. H., Wu S. V., Kim R. S., Martin M. G., Wong H. C., Mendelsohn J., Park J. G., Cuttittal F. Autonomous proliferation of colon cancer cells that coexpress transforming growth factor α and its receptor. Variable effects of receptor-blocking antibody. Gastroenterology, 102: 474-485, 1992.
Wu X., Fan Z., Masui H., Rosen N., Mendelsohn J. Apoptosis induced by an anti-epidermal growth factor receptor monoclonal antibody in a human colorectal carcinoma cell line and its delay by insulin. J. Clin. Investig., 95: 1897-1905, 1995.
Ciardiello F., Damiano V., Bianco R., Bianco C., Fontanini G., De Laurentiis M., De Placido S., Mendelsohn J., Bianco A. R., Tortora G. Antitumor activity of combined blockade of epidermal growth factor receptor and protein kinase A. J. Natl. Cancer Inst. (Bethesda), 88: 1770-1776, 1996.
Ciardiello F., Bianco R., Damiano V., De Lorenzo S., Pepe S., De Placido S., Fan Z., Mendelsohn J., Bianco A. R., Tortora G. Antitumor activity of sequential treatment with topotecan and anti-epidermal growth factor receptor monoclonal antibody C225. Clin. Cancer Res., 5: 909-916, 1999.
Bianco C., Bianco R., Tortora G., Damiano V., Guerrieri P., Montemaggi P., Mendelsohn J., De Placido S., Bianco A. R., Ciardiello F. Antitumor activity of combined treatment of human cancer cells with ionizing radiation and anti-epidermal growth factor receptor monoclonal antibody C225 plus type I protein kinase A antisense oligonucleotide. Clin. Cancer Res., 6: 4343-4350, 2000.
Perrotte P., Matsumoto T., Inoue K., Kuniyasu H., Eve B. Y., Hicklin D. J., Radinsky R., Dinney C.P.N. Anti-epidermal growth factor receptor antibody C225 inhibits angiogenesis in human transitional cell carcinoma growing orthotopically in nude mice. Clin. Cancer Res., 5: 257-265, 1999.
Petit A. M. V., Rak J., Hung M-C., Rockwell P., Goldstein N., Fendly B., Kerbel R. S. Neutralizing antibodies against epidermal growth factor and ErB-2/neu receptor tyrosine kinases down-regulate vascular endothelial growth factor production by tumor cells in vitro and in vivo. Am. J. Pathol., 151: 1523-1530, 1997.
Hasan S., Hassa P. O., Imhof R., Hottiger M. O. Transcription coactivator p300 binds PCNA and may have a role in DNA repair synthesis. Nature (Lond.), 410: 387-391, 2001.
You are going to email the following Enhanced Antitumor Activity of Anti-epidermal Growth Factor Receptor Monoclonal Antibody IMC-C225 in Combination with Irinotecan (CPT-11) against Human Colorectal Tumor Xenografts
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Canada extending ban on most foreign travellers to at least July 31
Ottawa plans to extend the sweeping travel ban that bars entry to all travellers who are not Canadian citizens or permanent residents, or Americans travelling for reasons deemed essential, for at least another month.
The order, which was set to expire tonight, "has been extended until July 31 for public health reasons," Rebecca Purdy, spokesperson for the Canada Border Services Agency, said in a statement to CBC News.
The order — one of a set of extraordinary new measures introduced on March 16 to stop the spread of COVID-19 — bans most foreign nationals from entering Canada if they arrive from a foreign country other than the U.S. (There are limited exceptions for air crew, diplomats and immediate family members of citizens. Some seasonal workers, caregivers and international students are also exempt.)
The current Order in Council — a cabinet decision made without having to go to Parliament — was set to expire at 11:59 p.m. ET Tuesday.
The government intends to continue with the blanket ban of foreign nationals from entering Canada at this time, officials confirm, rather than modifying the order to reopen the border to certain countries — for example, those with low infection rates or those allowing Canadian tourists to visit.
A separate order prohibits non-essential travel between Canada and the U.S. and is in effect until July 21 after being extended earlier this month. Under this order, essential workers, such as truckers and health workers, are allowed to cross the border.
Tuesday's decision by Ottawa to continue the ban on foreign nationals from entering Canada comes as the European Union agrees to allow some tourists to enter the EU beginning tomorrow. Canada is among the 15 countries on the EU's so-called "safe" list.
The EU said Tuesday it expects countries on that list to lift any bans they might have in place on European travellers.
Last week, Prime Minister Justin Trudeau pushed back against pressure to relax the ban on international travellers, arguing that moving too quickly could spark a second wave of the coronavirus.
"We are going to be very, very careful about when and how we start reopening international borders," Trudeau said at a briefing on June 22.
"I understand how difficult this is and how frustrating this is for some people, but we know that reopening too quickly or carelessly would lead us to a resurgence that might well force us to go back into lockdown, to shut down the economy once again, and nobody wants that."
A cabinet order requiring all travellers entering Canada to isolate for 14-days was set to expire Tuesday, but will be extended, a government official told CBC News on Monday. (Jennifer Gauthier/Bloomberg)
Quarantine order also to be extended: official
Tuesday marks six months since the World Health Organization was first informed of a cluster of unusual pneumonia cases in China – the first public indications of the coronavirus's emergence.
WHO Director-General Dr. Tedros Adhanom Ghebreyesus warned this week that the coronavirus pandemic is "not even close to being over" and that the outbreak is accelerating globally. To date, the WHO has recorded more than 10 million cases and 500,000 deaths globally.
CBC News reported on Monday that the federal government also intends to extend strict quarantine rules requiring travellers to isolate for 14 days upon their arrival in Canada. That separate order is also set to expire at 11:59 p.m. ET on Tuesday.
Under the Quarantine Act, travellers also need to confirm that they have a suitable place to isolate where they will have access to basic necessities, including food and medicine.
No one would be permitted to quarantine anywhere they could come into contact with vulnerable people. Those who, for example, normally live with an elderly person or someone with a compromised immune system would have to quarantine elsewhere.
If the Canada Border Services Agency suspects that a returning traveller is not going to comply with the rules, it can alert the Public Health Agency of Canada, which can then flag the RCMP's national operations centre. The RCMP has been playing a co-ordinating role with local police during the pandemic.
Maximum penalties for failing to comply with the Quarantine Act include a fine of up to $750,000 and/or imprisonment for six months. If someone jeopardizes another's life while wilfully or recklessly contravening the act, the penalties are even greater: $1 million or three years in prison, or both.
COVID-19 CANADÁ EUROPEAN UNION TRAVELLERS
Doctors are conducting 'dry runs' to decide who gets access to critical care if it gets to that point
Federal government lists COVID-19 testing services abroad after new rule strands many travellers
Ottawa 'actively looking' into loophole that allows quarantining travellers to get sickness benefit
TDSB needs millions to ensure safe return to school in fall, says it may have to cut French

redpajama_set_name: RedPajamaCommonCrawl


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You are here: Home / News / Community News / Features / Honoring healthcare workers and organizations — A behind-the-scenes look at the pandemic
Honoring healthcare workers and organizations — A behind-the-scenes look at the pandemic
By Mahlon Meyer
A ceremony honoring healthcare providers and organizations opened up new ways of looking at the pandemic. The virtual event, hosted by the Northwest Asian Weekly on Feb. 9, was structured so that supporters were free to pose any questions to the honorees.
The results were illuminating.
Healthcare professionals rated the Biden administration's handling of the crisis, shared previously unknown details about the origins of the outbreak in Seattle, reflected on what practices they hoped would continue, advised on how to find a balance between caring for others and oneself, and shared ways to overcome fear in the face of death.
In addition, Nate Miles, vice president of Strategic Initiatives at Eli Lilly, offered coaching about how to talk about an experiment undertaken by the United States government, lasting from the 1930s to the 1970s, to infect Black men with syphilis, the memory of which dissuades some Blacks from trusting the vaccine, he said.
"You've got to understand, you're going to have to slow walk these people and really convince them and you just can't gloss over it and pretend it didn't happen," he said.
John Chen emceed the Northwest Asian Weekly's virtual Healthcare Excellence Awards on Feb. 9
Miles said similar abuses experienced by other marginalized groups must also be acknowledged as part of a broader strategy to engender trust.
"You have to make this country understand what you did. You put people on reservations. You took people's homes from them and you put them in encampments and you want to call them camps. They weren't camps," he said, referring lastly to the incarceration of Japanese Americans during World War II.
Miles asked questions of Winona Hollins-Hauge, a social worker, activist, and radio host, and several other honorees, and his comments came in response to their remarks.
Asked to rate the Biden administration's handling of the pandemic, Hollins-Hauge praised the renewed "unleashing" of Dr. Anthony Fauci and said the new leadership "realized this is a big problem and that it is going to take everybody."
Another question from Miles, this time directed to Teresita Batayola, president and CEO of International Community Health Services (ICHS), asked her to compare differences between now and when the virus broke out.
As for the start of the pandemic, it has been largely unknown that it was in the Bellevue Clinic of ICHS where the first COVID-19 case was detected, even before King County Public Health announced it, she said.
Winnie Lee, the assistant medical director there, and one of the honorees, recognized COVID-19 even at that early stage and instantly ordered personal protective equipment (PPE) for herself and her staff.
As for the future, Batayola said one unanswered question was whether the Defense Production Act invoked by Biden would make a difference in the accelerated production of vaccines.
"The community needs to keep advocating to make sure that vaccines land in our hands instead of in mass distribution, where allocation formulas are so complex that community health centers like ICHS are being shorted," she said.
Responding to another question by Miles, Michael Byun, executive director at Asian Counseling and Referral Service (ACRS), praised his entire team and said it takes a collaborative approach to combat the pandemic.
"A lot of the solutions are within our communities, policymakers need to acknowledge that and lean in and listen to provide the support that the community is asking," he said.
Advocacy was also a theme of a question by Leeching Tran, vice president of the Viet-Wah Group, another event supporter.
"After the pandemic is over, what habits or behaviors do you want to see people continue or keep?" she asked Dr. Anthony Chen, director of health of Tacoma-Pierce County Health Department.
After offering praise for the widespread wearing of masks, social distancing, avoiding gatherings, and staying home while sick, all of which have actually lowered the transmission of the flu this season, Chen offered concrete hopes for the future. Asian countries routinely adopt mass mask wearing during cold and flu season, and he hoped we would consider doing so in this country.
Nevertheless, people should continue hygienic practices, self-care measures, such as eating healthily and getting enough rest, and continue to get outdoor exercise, he said. Finally, he marveled at the levels of compassion he had seen people showing for each other.
"We're all traumatized to some degree so this is wonderful that people are really focusing on compassion, I'd love to see that continue," he said.
Balancing compassion between oneself and others was the topic of a question by Tim Otani, director and external affairs manager at Union Bank.
"How do you train your students, our future nurses, to balance caring for patients with the need to take care of their own families?" he asked Gayle Robinson, a registered nurse and professional development specialist at Swedish Medical Center.
Beyond educating them in the appropriate use of PPE, Robinson, who also has a doctorate along with several other advanced degrees, teaches them to take into account how marginalized populations are impacted differently by society.
Albert Chun, International District branch manager for Washington Federal Bank, asked about the opposite end of the emotional spectrum.
"How do you overcome the fear of getting this disease while continuing to work in the health sector?"
Keith Koga answered, "Not just taking care of oneself physically and mentally and using PPE properly, 'but donning and doffing' it is vital." Koga is a clinical assistant nursing manager at Harborview, and manages a COVID testing site. "It's our job and responsibility to call anyone out that is not doing it correctly."
Min Chang, CEO of Kin On, added that both prevention and containment of positive cases were necessary. There is no good answer for how to take away the fear experienced by her staff, particularly in nursing, she said. But she has opened up units in Kin On's adult family home for nursing staff to stay in so they don't bring the virus home. Meanwhile, ACRS has provided classes about how to reduce stress.
Stress has reached new levels not only among healthcare workers, said Imei Hsu, a licensed mental health counselor and registered nurse, also an honoree. In comments made during the portion of the program supported by Rocky Fong, vice president of Corporate Social Responsibility at U.S. Bank, Hsu said with the dawn of 2021, we have moved beyond simply panic into disillusionment.
"In the disillusionment phase, there is more emphasis on what are you really going to do for your mental health now, because you've been putting it off saying, 'I think I can just get through this year (2020),' and that didn't quite happen the way we thought it would," she said.
One strategy she recommended was resilience.
"Never give up, always be available for others with the knowledge you have and the training you have." She also encouraged everyone to find inspiration from a past ancestor's story of courage.
Northwest Asian Weekly's associate publisher, John Liu, asked Yoon Joo Han, behavioral health director at ACRS, about tips for dealing with depression.
"Reach out to people you know and care about," said Han. "It's okay not to feel okay. Talk about your feelings with someone you trust."
Other questions centered on the logistics of vaccination, for instance, should those who've already had COVID-19 get the vaccine?
As long as you're feeling better and not still isolating, then yes, said Lee, the assistant director of the ICHS clinic in Bellevue.
While getting vaccinated, pop-up health clinics can also serve as places where a community can reunite and find emotional support.
A shot clinic last week at the Central Area Senior Center not only vaccinated 400 people, but it was the first time people had gathered in months.
"When we brought all those people together, with six feet of distancing and all the safety protocol, you wouldn't believe how good it felt, it felt like old home week," said Hollins-Hauge.
A less visible part of confronting the pandemic, however, emerged in remarks by Jim Doane, corporate counsel at Costco Wholesale. When large retail stores sell PPE from overseas, there is a herculean effort necessary to make sure it is reliable.
"You have to look at labeling, customs regulations, you have to look at FDA regulations, so there's a lot of nitty-gritty nerdy work behind the scenes to try to get non-counterfeit PPE into the country," said Doane.
Each individual honoree received a plaque, a bottle of red wine, and a gift certificate for a restaurant in the Chinatown-International District.
Organizations that were nominated received gift certificates from the Viet-Wah Supermarket.
"Please take time for yourself and don't forget to laugh every single day," said Assunta Ng, publisher of Northwest Asian Weekly.
Mahlon can be reached at info@nwasianweekly.com.
Filed Under: Features, Profiles, Community News, This week's issue Tagged With: 2021, VOL 40 NO 8 | FEBRUARY 20 - FEBRUARY 26


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To study neutrino interaction, Govt approves project to build India-based INO
News Bharati 12-Jul-2019
New Delhi, Jul. 12: The Government of India has approved a project to build the India-based Neutrino Observatory (INO) at Pottipuram in Theni District of Tamil Nadu. Briefly, the project aims to set up a 51000 ton Iron Calorimeter (ICAL) detector to observe naturally occurring atmospheric neutrinos in a cavern at the end of an approximately 2 km long tunnel in a mountain.
A neutrino detector is a physics apparatus which is designed to study neutrinos. Because neutrinos only weakly interact with other particles of matter, neutrino detectors must be very large to detect a significant number of neutrinos. Neutrino detectors are often built underground to isolate the detector from cosmic rays and other background radiation.
This will help to reduce the noise from cosmic rays that are ever present over-ground and which would outnumber the rare neutrino interactions even in a detector as large as ICAL.
Neutrinos are harmless and not till long ago were considered mass-less. Billions of neutrinos pass through our body every second, without hurting an atom or a molecule in our body. Two Nobel Prizes have already been given to scientists for research on these neutrinos. The neutrinos are part of elementary particle shower in natural cosmic rays and to isolate them require special observatories where they are secluded from other radiation.
This information was provided by the Union Minister of State (Independent Charge) Development of North-Eastern Region (DoNER), MoS PMO, Personnel, Public Grievances & Pensions, Atomic Energy and Space, Dr. Jitendra Singh in a written reply to a question in Rajya Sabha today.
The INO project does not disturb the ecosystem around the site and does not release any radiation, as it does not have any radioactive substance. It measures cosmic rays.
There is no other neutrino detector anywhere in India at present. ICAL at INO would be the first of its type.
India-based Neutrino Observatory neutrino detector Iron Calorimeter (ICAL) atmospheric neutrinos


Gingham Cambie dress | More blue fabric?
That picture is just amazing haha!
The dress looks lovely too, can definitely agree with the summer uniform connotation too.
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What is the challenge with employee disengagement? - Preparing you for the future of work.
What is the challenge with employee disengagement?
There is a significant body of research that reflects the degree of impact that employee disengagement has on the profitability of organisations.
First world culture conditions people to manage others in very specific kinds of unskilful ways and the anxiety created by stress and work pressure exacerbates this unskilfulness. This type of conditioning tends to dismantle employee's self-esteem promoting disengagement.
People tend to obtain a significant portion of their self-esteem from being valued and supported, and if this doesn't happen, they withdraw from interactions. They lose their spontaneity, creativity and motivation, the hallmarks of low engagement. Loss of productivity is the result.
Organisational cultures that promote over-professional behaviour tend to be cold, distant and uncaring, particularly under pressure, which is where the rubber meets the road. Messages delivered from management to staff in this manner, simply alienate them.
Managers often spend much of their time trying to inspire their staff to be more motivated and engaged; however if trust is not built first, inspiration will fail.
Coupled with this is the communication principle that most people need to be supported in their interactions, before they are challenged. If this concept is understood and practised, great trust can be built, which raises the self-esteem of staff, and engagement is catalysed.
When people don't know how to deal with their anxiety, their self-esteem crashes. Managers who become more skilled at mastering their own anxiety and raising their self-esteem, are in a better position to manage the self-esteem and hence disengagement of their subordinates.
The solution to this problem and the antidote to employee disengagement involves leaders and managers transforming their own self-esteem. They need to focus on their relationship with their own anxiety, and then communicating with their people in ways that raise self-esteem and engagement levels.
In summary, managers with high self-esteem know how to communicate with employees in ways that build the self-esteem and hence levels of engagement of their staff, raising productivity levels and reducing staff turnover.
Mark Kahn is a Practicing Clinical Psychologist and Management Consultant.


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October 2022 December 2022
Tue. 1 Nov, 2022
Tue. 1 Nov, 2022 6:00 pm
NOTICE IS HEREBY GIVEN that the Zoning Board of Appeals of the Town of Islip will hold a public hearing on Tuesday, November 1st, 2022 at the Town Hall West, 401 W Main Street, Islip, NY on the following applications at the times listed or as soon thereafter as they may be reached. At such time all interested parties will be given an opportunity to be heard. Any disabled person who needs a sign language interpreter or special accommodations to attend this meeting is asked to please contact Constituent Services at 631-224-5380 as early as possible in order for the Town to accommodate.
The meeting will also be streamed live over the Internet. Instructions on how to access the livestream will be available in the space below.
11/1/2022 - Zoning Board of Appeals Meeting Agenda
Mon. 7 Nov, 2022
Senior Fall Ball Returns November 7th, 2022
Mon. 7 Nov, 2022 11:00 am - 3:00 pm
Have a ball! Enjoy great food, music and more at the Town of Islip Senior's Fall Ball at the Timber Point Mansion. Tickets on sale now!
Wed. 9 Nov, 2022
Wed. 9 Nov, 2022 6:00 pm
NOTICE IS HEREBY GIVEN that the Planning Board of the Town of Islip will hold an in-person Public Meeting on November 9, 2022 at 6:00 pm, located at Islip Town Hall West, 401 W Main Street, Islip, NY 11751. The meeting will also be streamed live over the Internet. Instructions on how to access the livestream will be available in the space below.
Please note that due to the Novel Coronavirus (COVID-19) Pandemic and recent Legislation signed by the Governor, the format of this meeting is subject to change and may be held electronically via Zoom Webinar. In the event the format of this meeting changes to being held electronically instead of in-person, instructions on how to access an online livestream will be made available below. Attendees should check the Town website for instructions and any updates prior to the date of the meeting.
11/9/2022 - Planning Board Meeting Agenda
#1 Sayville Plaza Development LLC SP2022 066 Site Plan
#2 New Home holdings Corp CZ2022 026 Site Plan
#3 Veolia ES Technical Solutions, LLC CZ2022 027 Site Plan
#4 BDG SUN VET LLC CZ2022 028 Site Plan
#5 Central Islip Associates LLC CZ2022 015 Site Plan
#6 Vesta Acquisitions LLC CZ2022 016 Site Plan
#7 Julie & Chi Yim CZ2021 027 Site Plan
WATCH LIVE ON YOUTUBE
Thu. 10 Nov, 2022
Town Board Meeting (Budget Hearing)
Thu. 10 Nov, 2022 10:30 am
NOTICE IS HEREBY GIVEN that the Islip Town Board will hold a meeting on Thursday, Novemeber 10, 2022 at 10:30 a.m. at Town Hall, 655 Main St., Islip, New York and will be streamed live over the internet. Information regarding how to access the live stream will be available in the space below.
11/10/2022 - Town Board Budget Hearing Public Notice
Tue. 15 Nov, 2022
Town Board Meeting
Tue. 15 Nov, 2022 2:00 pm
NOTICE IS HEREBY GIVEN that the Islip Town Board will hold a meeting on Tuesday, November 15, 2022 at 2:00 p.m. at Town Hall, 655 Main St., Islip, New York and will be streamed live over the internet. Information regarding how to access the live stream will be available in the space below.
11/15/2022 - Town Board Meeting Agenda
11/15/2022 - Town Board Meeting Resolutions
NOTICE IS HEREBY GIVEN that the Zoning Board of Appeals of the Town of Islip will hold a public hearing on Tuesday, November 15th, 2022 at the Town Hall West, 401 W Main Street, Islip, NY on the following applications at the times listed or as soon thereafter as they may be reached. At such time all interested parties will be given an opportunity to be heard. Any disabled person who needs a sign language interpreter or special accommodations to attend this meeting is asked to please contact Constituent Services at 631-224-5380 as early as possible in order for the Town to accommodate.
11/15/2022 - Zoning Board of Appeals Meeting
Keep Islip Clean General Meeting
Receive information and updates on Keep Islip Clean activities such as the International Coastal Cleanups and other activities that focus on the beautification and maintenance of the roadsides throughout the Town of Islip.
There will also be a presentation on the Keep America Beautiful Recycling Pledge in honor of America Recycles Day, which is celebrated in November. Learn about volunteer opportunities to help end littering, beautify communities, increase recycling rates and educate youth.
For more information contact Keep Islip Clean at (631) 224-2627 or email This email address is being protected from spambots. You need JavaScript enabled to view it..
Change of Zone Hearing
Thu. 17 Nov, 2022 5:00 pm
NOTICE HEREBY IS GIVEN that there will be a Change of Zone hearing on Thursday, November 17, 2022 at 5:00 p.m. at Town Hall, 655 Main St., Islip, New York The meeting will also be streamed live over the Internet. Instructions on how to access the livestream will be available in the space below.
11/17/2022 - Change of Zone Public Notice Document
11/17/2022 - Change of Zone Hearing Agenda
Instructions to Join:
Wed. 30 Nov, 2022
Wed. 30 Nov, 2022 6:00 pm
NOTICE IS HEREBY GIVEN that the Planning Board of the Town of Islip will hold an in-person Public Meeting on November 30, 2022 at 6:00 pm, located at Islip Town Hall West, 401 W Main Street, Islip, NY 11751. The meeting will also be streamed live over the Internet. Instructions on how to access the livestream will be available in the space below.
11/30/2022 - Planning Board Meeting Agenda
#1 SP2022 020 Jason K Bria (Suncap Property Group, LLC) Site Plan
#2 PB2022 026 5150 Veterans LLC Site Plan
#3 PB2022 027 62 West Main Holding, LLC Site Plan
#4 PB2022 028 KCE NY 29, LLC Site Plan
#5 CZ2022 029 Craig Hinck Site Plan
#6 CZ2022 030 Costco Wholesale Corporation as successor by merger to Price Corporation Site Plan
#7 SP2014 059 Darshan Shah Site Plan


Today's and tonight's Melbourne, FL weather forecast, weather conditions and Doppler radar from The Weather Channel and Weather.com. Show me the weather in... city, zip, or place.
Weather Underground provides Melbourne weather radar information, Doppler detected storms and radar maps for all U.S. locations using Nexrad and TDWR radar stations.


When is the validation evaluated? It would seem that there would be exception thrown with the validation errors before it started to try and authenticate the current user/password. But it seems this is not the case.
The validation filter is executed in a Global Request Filter before the Service is executed. The [Authenticate] attribute has a -100 priority so it's executed before any Global Request Filters, it's the lowest priority of all built-in Request Filter Attributes so it's usually executed first.
Note: the recommended API to access the IAuthRepository is base.AuthRepository.
Right, when executed from a HTTP/MQ Request it should invoke the UpdatePasswordValidator.
Can you put a breakpoint on the RuleFor() line to see if it's being executed?
I found the issue. Turns out FluentValidation will pass when NewPassword is empty.


"A force to be reckoned with" is how, Canada's top Celtic publication, The Celtic Connection describes Pat Chessell, "...Check out his live show now so you could say you knew him when..."
Pat Chessell has been playing music for as long as he remembers. His first public performance was a guest spot with Belfast folk singer Danny Burns of the legendary Irish rebel group, The Wolfhound. Since then, Chessell has brought his music all over the world, performing his brand of Celtic influenced folk rock all over Canada and Ireland and most recently, a tour of New York. Wherever people have heard his music, they are always left wanting more due to his charasmatic stage presence, impressive guitar techniques and warm, character vocal stylings. John Murphy of the Boston based Irish punk e-zine, Shite 'N' Onions wrote: "Chessell is a strong vocalist... a name we will be hearing again." Pat is currently working on material for his second album due for release in 2010.


1. Newbies usually fail to keep track of EVERYTHING. Back up your passwords, transactions, time stamps, wallets, all for your own security and prevention in case you have any tax issues. Store them somewhere secure.
In the crypto world (most of the times) there is no one to reset or back up your information in case of loss or theft. If you lose it — you lose it forever. This is something that should be done from the moment you step into the cryptocurrency industry.
Ps: Learn about cryptocurrency taxes in your country of residency and stay on top of it because laws around this new technology are changing constantly, which is the reason why you should keep good track of every transaction you make, so you do not end up having to pay 300% of your crypto profits to any tax agency.
2. Live. Breath. And Eat All Crypto. Resist the urge to never leave the cryptocurrency world. Whenever we first learn about cryptos it is impossible to stay away from it, I admit it, once you understand the concept it is difficult not to indulge and be all in. But, the people around us may not be and it is important to become inclusive, share with others what you have learned about the ecosystem while keeping your feet on the ground. The world is not all about crypto. At least not yet.
3. The belief you will get rich overnight. This is a bad one guys. It is the same mistake many people make when they first get into stocks, day trading, etc. They all believe they will become trillionaires in a short period of time. That is a lie. Even if you see true stories about it, that is a very, I repeat, verysmall percentage of the people. Trading cryptos is hard work. This ecosystem requires all the same steps to success as with any other industry or market… Hard work is the path that could lead you to get rich.
4. Fear of Missing Out (FOMO) Once you start following the crypto groupies you will hear opinions about which coins to buy, or when to buy or sell coins ALL THE TIME, and such opinions could be right on the money, but you must know whether it fits with your plans or not. Wait, dear newbie do you even have a plan? We will talk about that a little further along.
For now, understand that new opportunities arise daily, so do not fear missing out. Rule Number #1 in trading is NEVER TRADE WITH YOUR EMOTIONS.
6. Lack of planning and patience. The most patient will be the most rewarded. This is true in stocks and it is true in the crypto world as well. Create a solid plan; do not just go shooting everywhere. Once you have a plan that fits your lifestyle and budget STICK TO THE PLAN. Resist the urge to move around or outside your trajectory. Try not to follow the crowds. And be patient during volatile times, eventually the ecosystem will stabilize.
7. Trusting others to care for you cryptos, including exchanges. This should be obvious, but you would be surprised at the number of people who entrusted their private information to others. Sometimes it is a necessary evil, I do agree that there´s a need for exchanges for example, but did you know you are giving them your private key? So please be careful which exchange you choose to "trust" and take advantage of all the security gadgets these exchanges offer (like 2FA).
Do not keep all your coins in a hot wallet or in exchanges, unless you are transacting them daily. Get a cold wallet for the coins you are hodling. And never-ever-ever-ever-ever-ever share your private keys or personal information with anyone or anywhere.
This ecosystem is an entirely new way of doing business, we must let go of our centralized mindset which once led us to trust others to keep us safe, and understand once and for all we make our own security in the crypto realm.
8. Not taking part in Communities or relying too much on them. Joining communities is something very important (you might want to read our blog post called "Cryptocurrency Communities: Why get involved?"), but you must not rely on them alone. Take a healthy part in them, cryptos need you to play your part in the community, but do not make your trades based solely on what is being said within the communities.
9. Not knowing your type of investor profile. Have you been watching the cryptocurrency market? How exactly do you feel about it? Do you own any cryptos? Did you lose money? Did it make you cringe? Or are you positive all this volatility will be worth it in the long run? Questions like these can determine what kind of investor you are (conservative, moderate or aggressive), and given that cryptocurrency is a market high of risks, not every profile, most likely conservatives, will fit well, unless they change their mindset about how to see the cryptocurrency ecosystem in the present moment.
An investor profile determines an individual´s preferences in investment decisions, it defines your investment style, which will lead you to understand what kind of investments is best suitable for you.
According to your profile, you can determine how to admin the cryptocurrency market. If you are moderate or aggressive, you will be fine with the volatility, but if you are a bit more conservative, you cannot approach the technology as an investment which you aim to make a profit, or else you will act out of fear at the first loss. As of now, any investor is actually investing in the technology; this is beyond a profit, cryptocurrency blockchain is the future as we know it, and that is what you are investing in, in the expansion of the technology.
Newbies must understand that in this early phase of the crypto market, there are no guarantees except for the technology. At this point, Moderate and Aggressive investors are okay with these risks because their profiles are inclined to focus in the long run even if they lose money for now. Conservatives, however, are freaking out lol.
If you would like to find out your investor profile, there are several suitability tests, search for one on google.
10. A tendency to form clans or to be a part of one. Leave your options open to new possibilities and opportunities. Resist the urge to divide into clans as most people do in the crypto world. You do not have to pick a side, diversify; understand the need for all sorts of projects, instead of seeing them as a threat to each other.
Cryptos are unique, they are not equal, and yet, they each serve a purpose. They can coexist in harmony ❤. But what do I know, ask Andrea Antonopoulos what he thinks, after all he is the expert.
Final Thought: Making mistakes is a natural part of any learning process, and though one could grow much from it, some slips can also cost you a lot of money especially in this industry.
We have all been through this process somehow; my only desire is to help newbies to make the best of their cryptocurrency and blockchain experience. We need educated participants, people who know what they are doing, and why they are doing it. If you know someone who could benefit from at least one point in this list, please make sure to share!
And if you have any other advice, please drop it in the comments!
Hope you have a successful journey!


Relocating into a new building comes with a share of its challenges. When you own a house, you should ensure that most of the installed systems are in their proper state. Below are the top repair works that can significantly improve the conditions is of your house.
The summer weather can be unforgiving to most people. You must ensure that you have an HVAC system that is properly functioning to regulate the heat. Frequent AC repairing in Dubai will guarantee that it will not break down during the hot weather. You should ensure that you hire Air Con repairs firm Dubai to ensure that the system is in the best shape. The lifespans of the air con can increase when you take serious care of it.
You need to check the pipes and ensure that they are properly functioning. The moist environment created by the leakage from water can lead to the growth of other microorganisms. You should ensure that all the pipes in the under the sink and the toilets are properly functioning. You can easily eradicate the molds and mildew when your house does not retain the moisture. You should get a Dubai plumber that will restore all the broken pipes to their correct status.
You should get a heat system expert that will check the insulators to ensure that they are in their best shape. You should ensure that the attic insulator is complete and find a new one if it is already damaged. You should never forget to repair the entire water heater systems. When the insulators are functioning effectively you will not worry about the weather in the winter and the summer.


It's that time of year. My inbox has been filing up with experts willing to share their health predictions for the coming year. Most are not particularly surprising: telemedicine, more apps for wellness, and so on. So I asked my community on Twitter to share some of its bold and/or outlandish predictions.
@chrissyfarr Beautiful, believable virtual humans for anxiety, mindfulness, and behavioral health. On smartphones, in VR/AR, everywhere.


text:
Articles filed under Charles Keeshan and Susan Sarkauskas
Does bail reform lead to more crime? What 2 Loyola University professors found
Jan 27, 2023 8:16 AM - Amid last year's contentious debate over the SAFE-T Act and the elimination of cash bail in Illinois, a pair of Loyola University Chicago criminal justice professors set out to determine whether bail reform does, as some claimed, lead to more crime. Here's what they found.
Guns in peanut butter and a whole chicken: What TSA agents found during a record-breaking 2022
Jan 20, 2023 5:00 AM - A record 6,542 firearms were seized at airport checkpoints across the country last year, and you won't believe where some of them were found.
Suburban cold cases solved and unsolved: How DNA has played a role so far
By Charles Keeshan And Susan Sarkauskas
Jan 13, 2023 8:19 AM - The Brown's Chicken murders isn't the only suburban cold case where DNA is credited with identifying a killer.
Lake County top prosecutor lobbies state legislators to pass assault weapon ban
Jan 6, 2023 5:00 AM - Lake County State's Attorney Eric Rinehart added his voice Thursday to those urging state legislators to pass a ban on so-called assault weapons before the legislative session ends next week.
The new search for Carol's killer: 47 years later, podcast investigates college student's slaying
By Charles Keeshan and Sussan Sarkauskas
Dec 23, 2022 3:03 PM - For nearly 50 years, the killing of 21-year-old college student Carol Rofstad has gone unsolved. But now, the Elk Grove Village woman's family has new hope she may finally get justice, with the release "Carol's Last Christmas," a true-crime podcast re-examining the case.
Botched police raids lead to change in state's attorney's warrants policy
Dec 16, 2022 5:30 AM - In the wake of some high-profile cases in which police served search warrants on the wrong location, the Cook County state's attorney's office is making some big changes in how it will require law enforcement to request, process and disclose information regarding warrants.
Once allies, Drew Peterson is silencing ex-attorney who wants to reveal 'what happened to Stacy'
Dec 9, 2022 5:00 AM - Once courtroom allies, Drew Peterson and defense lawyer Joel Brodsky have become legal adversaries. A state appeals court recently sided with Peterson and upheld a gag order on Brodsky, who had indicated in a TV interview that he was willing to spill the beans on his infamous client.
Misconduct or a team player? Naperville cop's firing winds up in federal court
Dec 2, 2022 5:00 AM - Was a veteran police officer fired this fall for misconduct involving falsified traffic stop information, or for failing to play along with a quota system he believes to be illegal?
State police's simulator lets cops practice responses to danger. We tried it, and it's hard.
Nov 25, 2022 6:23 AM - Police officers often say the public doesn't understand that sometimes situations go sideways in seconds. The Illinois State Police on Wednesday gave members of the media the chance to see just what that means and how its troopers practice for situations they may face.
Pair charged with fraudulently obtaining $100K in aid during pandemic
By Chuck Keeshan and Susan Sarkauskas
Nov 18, 2022 5:44 AM - A suburban duo are the latest state residents to face charges stemming from a monthslong state and federal investigation into unemployment insurance fraud during the COVID-19 pandemic.
Show 1-10 of 312 next »


When it comes to Light duty towing, it's never a good idea to call a run of the mill towing company. In fact, such companies are simply not equipped to handle RVs, large trucks, large 4X4s, pick up trucks, machinery, and vans. If you need to have these or other vehicles towed, and if you're located in Fayetteville, call Springdale Towing for a professional Light duty towing service. Our state of the art Light duty tow trucks can transport these and other vehicles from point A to point B quickly and efficiently. What's more, our tow truck drivers were trained in the ins and outs of Light duty towing and the safety precautions it entails. No matter where your vehicle broke down, we'll be available to offer you a top notch Light duty tow for the best price in town.
Greenland is a driver's city, and most of us spend a great deal of time on the road. No matter how carefully you drive, chances are you'll one day need to call a towing company for a towing service, When this happens, and this is especially true of you drive any of the Light vehicles mentioned above, it's crucial that you call a towing company that specializes in Light duty towing. That's where Fayetteville Towing comes in. Our Light duty tow truck drivers are on call all day every day, which means 24 hours a day, 7 days a week, and 365 days a year. Call Fayetteville Towing for an affordable Light duty tow. No matter where your car broke down and no matter how late it is (this includes weekends and holidays) we'll be ready to send out a state of the art Light duty tow truck.
Light duty towing has little in common with, say, flatbed towing. In fact, it's far more complicated. We train our licensed and insured tow truck drivers, all of whom were screened before being hired, in each and every aspect of Light duty towing and safety precautions. Using the very best tow trucks and winches, they'll tow your Light vehicle to the location of your choice in no time at all. Knowing from first hand experience how frustrating it is to wait for a towing company, we offer the best response time in Fayetteville: no more than 35 minutes. Even if it's raining, even if it's late at night, we'll be here to offer you the best Light duty towing service in the city.


Primary care is at the center of disease prevention, diagnosis and management. Dr. Gelfman practices membership-based medicine to provide exceptional access to help improve the overall patient experience.
Personalized preventative care including, but not limited to, dietary advice, treatment of elevated cholesterol and hypertension, smoking cessation, vaccines, colonoscopy screening, bone-density scans , as well as mammograms.
Admitting and inpatient care at Boca Raton Regional Hospital and a guarantee of no outside coverage.
Same-day or next-day appointments, 24/7 access via phone or text, appointments with specialists arranged by myself or my staff and on-site blood testing.
Charles Gelfman, M.D., is a Board-Certified Internist who has been practicing Internal Medicine for over 25 years. Dr. Gelfman is a Partner in the Boca Raton Physicians, P.A., a well-known and highly respected concierge practice.
Dr. Gelfman's unique, patient-centric approach to medicine, which has enabled him to cultivate relationships with patients and their family members spanning decades, combined with his volunteer efforts in the community, have earned him a reputation as a kind-hearted, old-fashioned community physician.
As a member of Dr. Gelfman's concierge practice, you will receive personalized care on an as-needed basis, with a specialized emphasis on preventive care.
If you would like to learn more about receiving personal, concierge care from Dr. Gelfman, please feel free to contact the office at 561-999-3043.


Discerning drivers will appreciate the 2017 GMC Acadia. A 3.6 liter V-6 engine pairs with a sophisticated 6 speed automatic transmission, and for added security, dynamic Stability Control supplements the drivetrain. A wealth of standard features means that you no longer have to sacrifice. Like all wheel drive, front and rear reading lights, a tachometer, variably intermittent wipers, heated seats, a power liftgate, cruise control, and much more. Third row seats provide an even greater maximum passenger capacity. Passenger security is always assured thanks to various safety features, such as: dual front impact airbags with occupant sensing airbag, front side impact airbags, traction control, brake assist, a security system, onStar, and 4 wheel disc brakes with ABS. It also arrives with a Carfax history report, providing you peace of mind with detailed information. We pride ourselves in the quality that we offer on all of our vehicles. Stop by our dealership or give us a call for more information.


text:
Stay a Little Longer
For the song by Brothers Osborne, see Stay a Little Longer (Brothers Osborne song).
"Stay a Little Longer"
Single by Bob Wills and His Texas Playboys
10-inch 78 rpm record
Songwriter(s)
Bob Wills, Tommy Duncan
"Stay a Little Longer" is a Western swing dance tune written by Bob Wills and Tommy Duncan. The title comes from a refrain in the chorus:
Stay all night, stay a little longer,
Dance all night, dance a little longer,
Pull off your coat, throw it in the corner,
Don't see why you can't stay a little longer.
The song consists of a number of unrelated verses, one of which (verse three) comes from an old folk song – "Shinbone Alley":
You ought to see my Blue Eyed Sally,
She lives way down on Shinbone Alley.
Number on the gate, the number on the door,
The next house over is the grocery store.[1]
Bob Wills and the Texas Playboys recorded it in 1945 and it reached number three in 1946.[2] Willie Nelson (number 22 in 1973) and Mel Tillis (number 17 in 1982) also charted Top 40 hits. The song has been recorded numerous times.
^ Cohen, Folk Music, p. 209-210: "A version of 'Shinbone Alley' was published in 1833, though its origins may be still older: "Old Miss Tuck and my aunt Sallie Both lived down in shinbone alley. No sign on the gate, no number on the door; Folks around here are gittin might poor."
^ Whitburn, The Billboard Book of Top 40 Country Hits, p. 493
Cohen, Norm. Folk Music: A Regional Exploration. Greenwood Press, 2005. ISBN 0-313-32872-2
Whitburn, Joel. The Billboard Book of Top 40 Country Hits. Billboard Books, 2006. ISBN 0-8230-8291-1
Willie Nelson singles
"No Place for Me"
"Man With the Blues" / "The Storm Has Just Begun"
"What a Way to Live"
"Nite Life"
"The Part Where I Cry" / "Mr. Record Man"
"Willingly"
"Touch Me"
"Wake Me When It's Over"
"Half a Man" / "The Last Letter"
"Am I Blue"
"I Never Cared For You"
"Pretty Paper"
"San Antonio Rose"
"I Love You Because"
"The Party's Over"
"Blackjack County Chain"
"Johnny One Time"
"Bring Me Sunshine"
"Fire and Rain"
"Yesterday's Wine"
"Stay All Night (Stay a Little Longer)"
"Bloody Mary Morning"
"Blue Eyes Crying in the Rain"
"If You've Got the Money I've Got the Time"
"Georgia on My Mind"
"Blue Skies"
"All of Me"
"Whiskey River"
"September Song"
"White Christmas"
"Help Me Make It Through the Night"
"My Heroes Have Always Been Cowboys"
"Midnight Rider"
"On the Road Again"
"Angel Flying Too Close to the Ground"
"Mona Lisa"
"I'm Gonna Sit Right Down and Write Myself a Letter"
"Always on My Mind"
"Let It Be Me"
"Last Thing I Needed First Thing This Morning"
"Little Old Fashioned Karma"
"Why Do I Have to Choose"
"Take It to the Limit"
"Without a Song"
"City of New Orleans"
"Forgiving You Was Easy"
"Living in the Promiseland"
"Heart of Gold"
"Spanish Eyes" (with Julio Iglesias)
"Nothing I Can Do About It Now"
"There You Are"
"Mr. Record Man"
"Ain't Necessarily So"
"Graceland"
"Mendocino County Line" (with Lee Ann Womack)
"Wurlitzer Prize" (with Norah Jones)
"Beer for My Horses" (with Toby Keith)
"The Harder They Come"
"You Don't Know Me"
"Cowboys Are Frequently, Secretly Fond of Each Other"
"Gravedigger"
"Roll Me Up and Smoke Me When I Die"
"Just Breathe"
"Grandma's Hands" (with Mavis Staples)
"It's All Going to Pot"
"Vote 'Em Out"
Book:Willie Nelson
Category:Willie Nelson
Collaboration albums
Notable singles
"Life Turned Her That Way"
"I Ain't Never"
"Good Woman Blues"
"Heart Healer"
"I Got the Hoss"
"What Did I Promise Her Last Night"
"I Believe in You"
"Ain't No California"
"Send Me Down to Tucson"
"Coca-Cola Cowboy"
"Blind in Love"
"Lying Time Again"
"Southern Rains"
"City Lights"
Pam Tillis
"Detroit City"
"Ruby, Don't Take Your Love to Town"
"Honey (Open That Door)"
This 1940s country song-related article is a stub. You can help Wikipedia by expanding it.
Retrieved from "https://en.wikipedia.org/w/index.php?title=Stay_a_Little_Longer&oldid=872220178"
Western swing songs
Bob Wills songs
Mel Tillis songs
Willie Nelson songs
Songs written by Bob Wills
Songs written by Tommy Duncan
1940s country song stubs
!!POLECAM!:
This page was last edited on 5 December 2018, at 23:19 (UTC).


There's an app for almost everything these days, whether it's shopping, tracking your exercise or finding your way. Now driver's licenses are making the transition from a card to a digital application on your mobile phone; a mobile driver license (mDL). Information such as change of address, over/under 21-years-old status, organ donor status and changes in driving status can be updated to the mDL and viewed in one instance.
mDL builds on the concept that at the start of every transaction, whether in the physical or digital space, lies a reliable, fraud-resistant identity document. The mDL allows all service providers- such as banks, courier services or governmental agencies - to check a person's identity on their mobile phone. Considering the increased focus on privacy, people are for example now able to confirm their passport number when opening a bank account without handing over the actual document. This will significantly speed up the onboarding process, allowing people to easily authenticate themselves for services without paper documentation or a physical interaction. Another market that can see dramatic effects from Mobile ID adoption is travel. E-tickets are already providing an easy and secure way of booking flight tickets and going through the e-gates using your mobile phone. When adding identity to the mix it would add further acceleration and security of the system.
However, rolling out a secure and interoperable mDL that can be used to confirm identity requires all use cases to be tested first. Moreover, all entities proofing IDs, including merchants, pharmacies, banks, police officers alike, need to be able to accept mDL to ensure smooth adoption. As standards become more apparent, it is mission-critical to harmonize intelligence and data sharing to create a seamless experience for both citizens and authorities while simultaneously empowering trust.
UL is uniquely positioned to support driving license issuers towards the introduction of an mDL. Our colleagues possess unique mDL knowledge and expertise through their leading involvement in the development of mDL globally. UL experts bring experience gained through their leadership role in the working group for international standardization of the mDL (ISO 18013-5) and their advisory position with regional associations of issuing authorities. This is complemented by hands-on experience building an mDL Proof-of-Concept, and involvement in testing mDL solutions. Also, UL coordinated the world's first "mDL Prototype Interoperability Party", a test event where the interaction between mDL and mDL Reader prototypes from multiple implementers was tried out. This knowledge and expertise is made available to our clients through Advisory services, Testing & Certification and Security Assurance.
UL's extensive experience in identity management and security supports the development, testing and compliance of mDL Issuance and Acceptance. Our global reach and independent view provides quality assurance to provide a secure, enhanced and trusted digital identity that creates a safer world.
To learn more, speak to a UL mDL expert now.


Menelik Watson has started just three Division I football games and is playing just his second season of football, but he's already making quite an impact as No. 4 Florida State's right tackle.
Watson played a season of college basketball at Marist and briefly tried boxing. After a year of junior college football in California, he signed with FSU. The 6-foot-6, 320-pound Watson graded out at 82 percent and had just one missed assignment as FSU accumulated 667 yards on offense in Saturday's 49-37 victory over Clemson.
"Menelik is a guy that is very hungry," FSU coach Jimbo Fisher said. "He's a guy that has a lot of ability and is progressing well. He learns well. He has the physical ability and the natural instinct to play, and he's developing himself into a very good player."
Tough ticket: Saturday's game at South Florida has proven to be one of the toughest tickets for a regular-season road game in recent memory for FSU fans.
"Just the fact that it's our demographics, playing in Tampa," said FSU assistant athletics director Ben Zierden, who oversees the ticket office. "Obviously, we have a huge, loyal fan base down there."
FSU exhausted its allotment of 10,500 tickets this summer. Some FSU fans that were not able to buy tickets as part of FSU's allotment have purchased USF's season tickets just to guarantee themselves a seat on Saturday.
Tampa's Raymond James Stadium should be host to a capacity crowd of approximately 65,500 fans Saturday night – but possibly more. The largest USF home crowd was in 2007 when the No. 18 Bulls upset No. 5 West Virginia before 67,012 fans.
There were about 700 FSU-USF tickets available on Stubhub's website on Tuesday afternoon (starting at $90 per ticket). No tickets were available on Ticketmaster's website.