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Benzothiazole derivatives are known for various biological activities, and their potency in cancer therapy has received considerable attention in recent years. YLT322, a novel synthesized benzothiazole derivative, exhibits potent anti-tumor activity via inducing apoptosis both in vitro and in vivo. In this study, we found that YLT322 showed growth inhibition against a broad spectrum of human cancer cells and induced apoptosis of HepG2 cells in a dose- and time-dependent manner. The occurrence of its apoptosis was associated with activation of caspases-3 and -9, but not caspase-8. YLT322 increased the expression of Bax, decreased the expression of Bcl-2, and induced the release of cytochrome c which activates the mitochondrial apoptotic pathway. The down-regulation of phosphorylated p42/44 MAPK and phosphorylated Akt was also observed. Moreover, YLT322 suppressed the growth of established tumors in xenograft models in mice without obvious side effects. Histological and immunohistochemical analyses revealed an increase in TUNEL and caspase-3-positive cells and a decrease in Ki67-positive cells upon YLT322. These results suggest that YLT322 may be a potential candidate for cancer therapy.
Citation: Xuejiao S, Yong X, Ningyu W, Lidan Z, Xuanhong S, Youzhi X, et al. (2013) A Novel Benzothiazole Derivative YLT322 Induces Apoptosis via the Mitochondrial Apoptosis Pathway In Vitro with Anti-Tumor Activity in Solid Malignancies. PLoS ONE 8(5): e63900. https://doi.org/10.1371/journal.pone.0063900
Editor: Kalpana Ghoshal, The Ohio State University, United States of America
Received: November 21, 2012; Accepted: April 8, 2013; Published: May 30, 2013
Copyright: © 2013 Xuejiao et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by Shijiazhuang Pharmaceutical Group Co., Ltd. (project name SKLB163; contract number 11H0684). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: This work was supported by Shijiazhuang Pharmaceutical Group Co., Ltd. (project name SKLB163; contract number 11H0684). There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.
Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the third cause of cancer-related death [1], [2]. Chemotherapy is the most usual treatment approach, in addition to resection and liver transplantation; however, it does produce satisfactory results because of the resulting poor response rates, severe toxicities and high recurrence rates [3], [4]. Resistance to apoptosis might account primarily for the resistance of tumor cells to chemotherapy and for cancer progression [3], [5]. Modulation of apoptosis sensitivity of cancer cells has emerged to be a promising strategy to induce cell death in cancer cells and indeed, most chemotherapeutic drugs could tumors by triggering cancer cell apoptosis [6].
Generally, drugs induce apoptosis in cancer cells through two pathways: cell death receptor-mediated extrinsic pathway and mitochondrial-mediated intrinsic pathway. In the extrinsic pathway, the ligation of so-called death receptors results in the activation of the protease caspase-8 which then cleaves and activates downstream effectors caspase-3 and/or -7, resulting in chromatin condensation, DNA degradation, cell shrinkage and formation of apoptotic bodies [7]. In the intrinsic pathway, Bcl-2 family members are the key regulators of apoptosis. Once the anti-apoptotic members of this family are inhibited and/or the pro-apoptotic members are activated, mitochondrial integrity is disrupted and cytochrome c is released. As a consequence of these changes, cytochrome c interacts with the Apaf-1 (apoptotic protease activating factor - 1) and ATP, and then binds to pro-caspase-9. This interaction results in the cleavage of pro-caspase-9, which in turn activates the effector caspase-3 and/or -7 [8]. Various anti-cancer agents have been shown to induce apoptosis through the intrinsic pathway [9].
Because of their wide range of biological activities, a number of benzothiazole derivatives have attracted interest for their potential pharmacological applications [10]–[12]. In recent years, extensive study has focused on assaying novel benzothiazole derivatives for anti-tumor activities. Our research group has been interested in the design, synthesis, screening and biological evaluation of novel benzothiazole derivatives as potential anticancer agents. Among these, 2-Chloro-N-(2-(2-(5-methylpyridin-2-ylamino)-2-oxoethylthio)benzo[d]thiazol-6-yl) acetamide (YLT322) displays robust anti-proliferative activity in vitro [13]. In this study, we demonstrated that YLT322 can induce apoptosis in human hepatocellular carcinoma cells via the mitochondrial apoptotic pathway and the down-regulation of phosphorylated Akt/MAPK, and also inhibit tumor growth in vivo by inducing apoptosis.
Materials and Methods
Drugs and reagents
2-Chloro-N-(2-(2-(5-methylpyridin-2-ylamino)-2-oxoethylthio)benzo[d]thiazol-6-yl) acetamide (YLT322) (Fig. 1) was synthesized previously by our group (State Key Laboratory of Biotherapy, Sichuan University, Sichuan, China) [13] and the structure was confirmed by 1H-NMR, 13C-NMR and HRMS (ESI). Purity (98%) was measured by HPLC analysis. YLT322 was dissolved in dimethyl sulfoxide (DMSO) at a stock concentration of 10 mM and stored at -20°C. For all in vitro assays, the working dosage was freshly diluted in relevant medium with a final DMSO concentration of less than 0.1%.
3-(4,5)-dimethylthiahiazo(-z-y1)-2,5-di-phenytetrazolium bromide (MTT), Rhodamine-123 (Rh123), Hoechst 33342, dimethyl sulfoxide (DMSO) and propidium iodide (PI) were purchased from Sigma Chemical Co. (St. Louis, MO). Annexin V-FITC apoptosis detection kit was purchased from Roche (Indianapolis, IN). The antibodies against caspase-3, caspase-9, Bax, Bcl-2, cytochrome c, Akt/p-Akt, p44/42 MAPK (mitogen-activated protein kinase)/p-p44/42 MAPK, and COX-4 were purchased from Cell Signaling Technology Company (Beverly, MA). Antibody against caspase-8 was from Thermo Scientific (Breman, Germany) and antibody against β-actin was obtained from Santa Cruz Biotechnology Company (Santa Cruz, CA). TUNEL (the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling) assay kit was purchased from Promega Company (Madison, WI). Z-VAD-FMK (Caspase Inhibitor), LY294002 (PI3K/AKT Inhibitor), PD98059 (MAPK Inhibitor) were purchased from Beyotime (Beijing, China) and Ac-LEHD-FMK and Ac-IETD-FMK were from Keygen (Nanjing, China).
Cell culture
The human HCC SMMC-7721,Bel-7402 and Bel-7404 cell lines were obtained from the China Center for Type Culture Collection (CTCCC,Wuhan, China). All other cell lines were acquired from the American Type Culture Collection. Cells were propagated in DMEM or RPMI 1640 medium (Gibco BRL, Grand Island, N.Y.) supplemented with 10% fetal bovine serum (FBS; Gibco, Auckland, N.Z.), 100 units/mL penicillin and 100 units/mL streptomycin under humidified conditions with 5% CO2 at 37°C. No further authentication was conducted for tumor cell lines. Cells without YLT322 treatment served as a vehicle group.
Cell proliferation assay
Cell viability after YLT322 treatment was performed by the MTT assay. Briefly, the exponentially growing cells (3–5×103 cells/well) were seeded in 96-well plates and cultured for 24 hours. After treatment with various concentrations of YLT322, a volume of 20 µl of MTT solution (5 mg/ml) was added to each well and incubated for another 2–4 hours at 37°C. Then the medium was discarded and the formazan salt was dissolved with 150 µl DMSO for 15–20 minutes. The absorbance of each well was measured with Spectra MAX M5 microplate spectrophotometer (Molecular Devices) at 570 nm wavelength, and the median inhibitory concentration (IC50) of each cell line was calculated. Three replicate wells were used for each analysis. The results were obtained from three separate experiments.
Colony formation assay
To test the survival of HepG2 treated with YLT322, the cells were plated (6–10×104 per well) in a six-well plate and incubated overnight at 37°C. After 48 h exposure to various concentrations of YLT322, the cells were cultured for another 12 days with fresh medium and subjected to a clonogenic assay as previously described [14].
Morphological analysis after Hoechst staining
Morphological changes associated with apoptosis in HepG2 cells were detected by Hoechst 33342 staining. Briefly, the cells were plated onto 18-mm cover glass in 6-well plates for 24 h and then treated with YLT322 for another 24 h. After treatment, cells were rinsed with cold PBS and fixed in paraformaldehyde solution for 20 minutes. The cells were stained with Hoechst 33342 solution (5 u g/ml) followed by PBS washing and examination under fluorescence microscope (Zeiss, Axiovert 200, Germany) to identify the nuclear morphology of apoptotic cells.
Apoptosis analysis by flow cytometry (FCM)
To further confirm the apoptosis inducing effect of YLT322, Annexin V-FITC apoptosis detection kit was used. Briefly, after treatment with different concentrations of YLT322 for 12, 24 and 48 h as described above, cells were harvested and washed with cold PBS twice. After centrifugation, cells were stained with Annexin V-FITC and PI, and then analyzed with FCM (Becton–Dickinson, USA).
Furthermore, to confirm whether caspases, AKT and p44/42 MAPK are involved in YLT322-induced apoptosis in HepG2 cells and to determine which pathways are important, we treated cells with or without 2 µM YLT322 combined with 20 µM Z-VAD-FMK (cell-permeable caspase inhibitor), 50 µM Ac-LEHD-FMK (caspase-9 inhibitor), 50 µM Ac-IETD-FMK (caspase-8 inhibitor), 50 µM LY294002 (PI3K/AKT Inhibitor) or 50 µM PD98059 (MAPK Inhibitor). Annexin V/PI staining was analyzed by FCM. We also studied the apoptosis inducing effect of YLT322 in other hepatocellular carcinoma cells, including Bel-7402, Bel-7404 and SMMC-7721 by PI staining.
Mitochondrial membrane potential (ΔΨ) assay
Changes in mitochondrial transmembrane potential (ΔΨ) were evaluated by staining cells with Rh123 as described previously [15]. Cell culture and drug treatment were done as described above. The harvested HepG2 cells were washed with cold PBS after incubation with Rh123 (5 µg/ml) at 37°C for 30 min in the dark and then measured by flow cytometry .
Western blot analysis
Cells were lysed in RIPA buffer after treatment with YLT322 for 48 h and the lysates were centrifuged at 13,000 g for 15 min at 4°C. The supernatant was harvested and the protein concentration was measured by the Lowry method. Equal amounts of total proteins were subjected to SDS-PAGE and transferred onto polyvinylidene fluoride membranes. After electrophoresis, the membranes were blocked for 1.5 h at room temperature and incubated overnight at 4°C with the respective primary antibodies followed by the secondary antibody conjugated to horseradish peroxidase. The immunostaining signal was detected by the enhanced chemiluminescence system (Amersham, Piscataway, NJ).
Subcutaneous xenograft models
The study procedures were approved and conducted in accordance with the Animal Care and Use Committee of Sichuan University. 100 µL tumor cell suspension containing between 5×106 and 1×107 cells were injected subcutaneously into the right flank of the seven-week-old female BALB/c athymic nude mice. When tumors reached an average volume of 100 mm3, the mice were randomly divided into groups of 5 to 6. Animals were given YLT322 (37.5–150 mg/kg) or vehicle once daily by intraperitoneal injection. Tumor size and body weight were measured every three days, and clinical symptoms were observed daily. The tumor volume was calculated according to the following formula:V = (length×width2/2).
Immunohistochemistry and TUNEL analysis
After treatment for seven days, tumors of HepG2 models were removed, fixed, routinely processed and embedded in paraffin. The sections containing tumors were stained with H&E or treated with specific antibodies for immunohistochemistry analysis. Apoptotic cells in the tumor tissue were identified by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining using an apoptotic cell detection kit.
Sub-acute toxicity test
A sub-acute toxicity test was performed in BALB/C mice by oral administration with 2 g/kg YLT322. The clinical symptoms of the animals, including mortality, body weight and movement, were observed once a day for 14 days. Blood was obtained for hematological and serum biochemistry analysis by Hitachi 7200 Blood Chemistry Analyzer and a Nihon Kohden MEK-5216K Automatic Hematology Analyzer.
YLT322 inhibited proliferation of cancer cell lines
In order to determine whether YLT322 possesses the potential to be an effective anti-cancer agent, we first examined whether YLT322 exerts a growth inhibition effect on cancer cells by treating a panel of 24 established cancer cell lines of different histotypes with YLT322 for 48 hours and then assaying cell viability by MTT assay. YLT322 exhibited a significant cell viability inhibition with an IC50 between 0.39 µM and 7.70 µM, and had a more potent proliferation inhibitory effect than the positive control doxorubicine in 14 of 24 cell lines (Table 1). Since human hepatocellular cancer cell line was most sensitive to YLT322, we chose this cell line for further experiments. The viability of the human hepatocellular cancer cell lines exposed to increasing concentrations of YLT322 up to 5 µM for 24–72 hours was attenuated with increasing concentration and duration of exposure (Fig. 2A). These data directly suggest that YLT322 exerts a more potent growth inhibitory effect than doxorubicine and can inhibit hepatic carcinoma cell proliferation in a concentration- and time-dependent manner. In addition to these experiments, we have also examined the effects of YLT322 on cell viability using clonogenic assays as described below.
A. Cells (a:HepG2; b:Bel-7402; c: Bel-7404;d:SMMC-7721) were treated with concentrations of YLT322 ranging from 0.15 to 5.0 µM, for 12 to 48 hours and cell viability was determined by MTT assay . Data are expressed as mean ± SD. for at least 3 independent experiments. (*p<0.05; **p<0.01; ***p<0.001) B. Effects of varying concentrations of YLT322 on colony formation of HepG2 and Bel-7402 cells after two weeks treatment. C. Statistical results of colony-forming assays presented as surviving colonies (percentage of untreated control). Data are expressed as mean ± SD. for at least 3 independent experiments. (*p<0.05; **p<0.01; ***p<0.001).
Inhibition of cell growth/survival by clonogenic assay
To further determine the effect of YLT322 on cell growth, we conducted clonogenic assay after YLT322 treatment. Fig. 2B and 2C present a significant inhibition of colony formation of HepG2 and Bel-7402 cell lines in a concentration-dependent manner after exposure to YLT322 compared with vehicle. When YLT322 concentration was at 1 µM, nearly no colony formation was detected. Overall, the results from clonogenic assay were consistent with the MTT data as shown in Fig. 2A, suggesting that YLT322 can inhibit cell growth in HepG2 and Bel-7402 hepatocellular cancer cell lines.
Induction of apoptosis by YLT322
We next studied the induction of apoptosis in hepatocellular cancer cells treated with YLT322 to determine whether inhibition of cell viability is associated with activation of the programmed cell death pathways. As Fig. 3A and B indicates, YLT322 treatment increased the percentage of Sub-G1 cells from 3.2% in non-treated group to 88.5% in 2 µM YLT322-treated group in HepG2 cells. Similarly, in other hepatocellular cancer cell lines such as Bel-7402, Bel-7404 and SMMC-7721, the number increased from 1.8% to 55.3%, from 1.1% to 24.6%, and from 4.2% to 20.8%, respectively. Furthermore, the HepG2 cells exhibited features of apoptotic cells as revealed by Hoechst 33342 staining, including bright-blue fluorescent condensed nuclei, reduction of cell volume and nuclear fragmentation (Fig. 3C).
A. Flow cytometric analysis of PI-stained HCC cell lines, including HepG2, SMMC-7721, Bel-7402 and Bel-7404, after treatment with 2 µM YLT322 for 48 hours. B. Statistical results of apoptosis assays presented as surviving cells (percentage of untreated control). Data are expressed as mean ± SD. for at least 3 independent experiments. (*p<0.05; **p<0.01; ***p<0.001). C. Fluorescence microscopy analysis of Hoechst 33342-stained HepG2 cells after incubation with varying concentrations of YLT322 (0 µM (a), 0.5 µM (b), 1 µM (c), 2 µM (d)) for 24 hours (×40). D. Flow cytometric analysis of cells stained with Annexin V-FITC/PI after treatment with various concentrations (0 µM, 0.5 µM, 1 µM, 2 µM) of YLT322 for 12 to 48 hours.
To confirm this cell death result, we also used other methods to detect apoptosis: Annexin V – FITC and PI fluorescence staining was analyzed by flow cytometry. Fig. 3D shows the time- and dose-dependent changes in the percentage of apoptotic cells when cells were exposed to 0.5 µM, 1 µM or 2 µM YLT322 for 12, 24 and 48 hours. After 48 hours, we found an obvious concentration-dependent reduction in the percentage of surviving cells (from 81.1% at 0.5 µM, to 66.7% at 1 µM, to 45.1% at 2 µM), with nearly a 40% change between the lowest and highest concentration. However, the earlier time points did not present significant changes, which was consistent with the MTT data as shown in Fig. 2A.
Effect of YLT322 on the intrinsic apoptosis pathway
Apoptosis is associated with the activation of specific caspase cleavage cascades. To further characterize the apoptosis pathways stimulated by YLT322, we analyzed the proteolytic processing of caspase-3, caspase-8 and caspase-9. A reduction in pro-caspases-9 and -3 and an increase in the levels of their cleaved forms were observed following exposure to YLT322 for 48 hours in HepG2 and Bel-7402 cells. The effect on the level of pro-caspase-8 protein was also observed, but the increase in cleaved-caspase-8 was not significant (Fig. 4A). To explore whether YLT322-induced apoptosis is specifically associated with caspase activation and to determine which type of apoptotic pathways is predominant, we examined by FCM whether Z-VAD-FMK (a general caspase inhibitor), Ac-LEHD-FMK (caspase-9 inhibitor) and Ac-IETD-FMK (caspase-8 inhibitor) can affect the extent of YLT322-induced apoptosis. As shown in Fig. 4B, compared with YLT322 treatment alone, treatment with 2 µM YLT322 combined with 20 µM Z-VAD-FMK decreased the percentage of apoptotic cells from 46.2% to 26.9% while that with 50 µM Ac-LEHD-FMK decreased the percentage of apoptotic cells to 22.6%, . However, the combination of YLT322 with 50 µM Ac-IETD-FMK had only a slight effect on YLT322-induced apoptosis. These results indicate that the caspase family of proteins plays an important role in YLT322-induced apoptosis, and that the intrinsic pathway contributes more to YLT322-induced apoptosis than the extrinsic pathway.
A. The expression levels of pro-caspase-8, -3, -9, and their activated forms in HepG2 and Bel-7402 cells treated with YLT322 for 48 hours as assayed by western blot. B. HepG2 cells were treated with 2 µM YLT322 alone or in combination with Z-VAD-FMK (a general caspase inhibitor), Ac-LEHD-FMK (caspase-9 inhibitor) or Ac-IETD-FMK (caspase-8 inhibitor). Data are expressed as mean ± SD. for at least 3 independent experiments. (*p<0.05; **p<0.01; ***p<0.001). C. The mitochondrial membrane potential (ΔΨ) was detected by RH123 staining after treatment with YLT322 at varying concentrations of 0 µM (Dark blue), 0.5 µM (blue), 1 µM (orange), 2 µM (green) for 12 to 48 hours (Bars, SD; Column, mean; n, 3;*p<0.05; **p<0.01; ***p<0.001). D. The level of cytochrome c in the cytosol and mitochondrial fraction was analyzed by western blot analysis after incubation of cells with YLT322 for 48 hours. Expression of COX-4 served as the loading control for the mitochondria fraction. E. The protein levels of Bcl-2 and Bax were detected by western blot in HepG2 cells treated with YLT322 for 48 h. F. The mRNA level of Bax was examined by real-time RT-PCR in HepG2 cells treated with YLT322 for 48 h. (Bars, SD; Column, mean; n, 3;*p<0.05; **p<0.01; ***p<0.001). G. The expression levels of Akt, p44/42 MAPK and their phosphorylated forms were assayed by western blot in HepG2 cells treated with YLT322. H. HepG2 cells were treated with 2 µM YLT322 alone or in combination with LY294002 (PI3K/AKT inhibitor) or PD98059 (MEK/ERK inhibitor). Data shown are representative of three independent experiments.
Effects of YLT322 on cytochrome c and ΔΨ
A limiting step in the intrinsic apoptotic pathway is the damage of mitochondria and the release of cytochrome c from mitochondria into the cytosol. To observe the change in ΔΨ in cells exposed to YLT322, a mitochondria-specific and voltage-dependent dye RH123 was used. A time- and dose-dependent reduction in ΔΨ was observed when cells (HepG2) were exposed to 0.5 µM, 1 µM, or 2 µM YLT322 for 12, 24, and 48 hours (Fig. 4C). After 48 hours, there was a significant dose-dependent reduction in the ΔΨ of more than 40.0% in the range of 0.5 to 2 µM. However, at 12 and 24 h, the loss of ΔΨ was indiscernible, which was in line with the Annexin V – FITC/PI data. Protein analysis showed that after YLT322 treatment for 48 h, cytochrome c level was increased in the cytosolic fractions but decreased in the mitochondria fraction (Fig. 4D).
Effect of YLT322 on the expression levels of Bax and Bcl-2
Bcl-2 family proteins are essential for regulating mitochondrial integrity through the balance between anti-apoptotic and pro-apoptotic members [16]. We examined by western blot analysis the expression of some Bcl-2 family proteins in HepG2 cells after YLT322 treatment for 48 hours. As shown in Fig. 4E, the expression of Bcl-2 significantly decreased in a concentration-dependent manner while that of Bax was increased. Furthermore, real-time reverse transcription- PCR (Fig. 4F) revealed that the Bax mRNA was up-regulated by YLT322, which was consistent with the result of western blot analysis in Fig. 4E.
Effect of YLT322 on p44/42 MAPK and Akt signaling pathways
We next investigated whether p44/42 MAPK and Akt, which are considered to be important for cell proliferation and apoptosis, are involved in YLT322-mediated apoptosis [17]. We found that YLT322 decreased the expression of phosphorylated Akt and phosphorylated-MAPK without affecting their total expression level (Fig. 4G). Next, cells pretreated with the PI3K/AKT inhibitor LY294002 and MEK/ERK inhibitor PD98059 were exposed to 2 µM YLT322 for 48 hours. As shown in Fig. 4H, LY294002 and PD98059 reduced YLT322-induced cell apoptosis from 45.8% to 22.2% and to 26.1%, respectively. This suggests that p44/42 MAPK and Akt play an important role in YLT322-induced apoptosis.
Anti-tumor efficacy of YLT322 in human tumor xenograft models
To assess the antitumor effect of YLT322 in vivo, HepG2 and HCT116 cells were injected subcutaneously into the right flanks of BALB/c nude mice to establish xenografts. Tumor-bearing mice were administered with vehicle or YLT322 at a dosage of 50, 100 or 150 mg/kg/day for the HepG2 model and 37.5, 75 or 150 mg/kg/day for the HCT116 model. YLT322 substantially suppressed tumor growth in a dose-dependent manner, with the inhibition of tumor progression at 66.7% in the HepG2 model treated with 150 mg/kg YLT322 and at 56.7% in the HCT116 model treated with 150 mg/kg YLT322 (Fig. 5A,C). Furthermore, YLT322 treatment was well tolerated and did not cause significant loss in body weight (Fig. 5B,D).
Animals were given YLT322 or vehicle by intraperitoneal injection once daily (qd) at the indicated dosage levels. The tumor volume and animal weight of the HepG2 (A and B) and HCT116 (C and D) xenograft models were measured twice weekly. Points, mean tumor volume (mm3); bars, SE; n, 6; *p<0.05; **p<0.01; ***p<0.001. (E and F) After 6 days of YLT322 treatment, the HepG2 tumors treated with 150 mg/kg YLT322 or vehicle were collected separately (three per group). The sections containing tumors were examined by immunohistochemical analysis with anti-Ki67 (×40) and caspase-3 antibody (×40), and by hematoxylin staining (×5). TUNEL detection was performed for apoptosis analysis (×20). G. The percentage of apoptosis in each group. TUNEL-positive cells were counted in five high power fields/slide, and data were summarized in terms of percent positive cells. Bars, SEM; columns, mean; *p<0.05; **p<0.01; ***p<0.001.
To further demonstrate that tumor growth inhibition in vivo is a result of apoptosis, histological and immunohistochemical analyses were performed on tumor tissues isolated from HepG2 tumor model. As shown in Fig. 5E, 5F, hematoxylin and eosin staining revealed a significant regression of tumors isolated from YLT322-treated animals which was not observed in the vehicle group. Meanwhile, YLT322 treatment caused a significant decrease in the number of Ki67-positive cells, but an increase in the number of caspase-3-positive cells. These changes were not observed in the vehicle group. The percentage of TUNEL-positive cells in YLT322-treated tumors was higher (36.6%) than that in the vehicle group(7.6%)(Fig. 5G). Taken together, these results clearly show that YLT322 inhibits tumor growth in vivo through a reduction in proliferating cells and increased apoptosis in human tumor xenograft model.
Toxicity Evaluation
For a preliminary safety estimate of YLT322, a sub-acute toxicity test was performed. No significant change was observed in hematological and serum biochemical values (Fig. 6A) and body weight (Fig. 6B) in YLT322-treated mice compared to control.
A. Hematological and serum biochemical values of mice at day 13 (n = 5 for both control and treated). B. Data showing average body weight changes in the control (n = 5) and YLT322 treated mice (n = 5). Values are indicated in meanSEM.
Dysregulation of the programmed cell death process (apoptosis) is a hallmark of many types of malignancies and the cause of cancer cell resistance to chemotherapy [7], [18], [19]. Thus restoring apoptosis is a promising strategy for effectively treating cancers [20]. Currently, many therapeutic approaches, including various biologicals, hormones, and small molecule chemotherapeutic agents such as ABT737, AT101, HS-113 and chloroquine, inhibit tumors by triggering cell apoptosis [18], [21]–[24]. Previous studies have demonstrated that some benzothiazole derivatives can induce apoptosis [10], [13], but the mechanism has not been clearly determined. In the present study, our results show that YLT322 is a broad spectrum anti-cancer compound with high apoptotic-inducing activity to a panel of cancer cells with the most potent effect observed with HepG2. Moreover, the anti-cancer effect of YLT322 was demonstrated to be associated with the induction of apoptosis.
Possible drug targets for regulating cell apoptosis have been widely studied [7]. In general, the action of apoptosis is through two very distinct signaling pathways: intrinsic pathway or extrinsic pathway [25], [26]. Caspase-3, the major effector caspase, is responsible for the cleavage of the majority of polypeptides that undergo proteolysis in an apoptotic cell and plays a central role in both types of apoptotic pathways [27]. It is activated by upstream effector proteins including caspase-8 and caspase-9, the apical proteases in the extrinsic and intrinsic pathways, respectively [7], [28]. After YLT322 treatment, we observed in cells a reduction in procaspase-3, -8 and -9 and a concomitant increase in their cleavage, indicative of enzyme activation,. Addition of the irreversible inhibitors, Z-VAD-FMK (caspase inhibitors) and Ac-LETD-FMK(caspase-9 inhibitors) significantly decreased the percentage of apoptotic cells after treatment with YLT322, suggesting that cell apoptosis induced by YLT322 may be dependent on caspase activation. However, Ac-IETD-FMK, a caspase-8 inhibitor, was unable to antagonize YLT322-induced cell death. It is possible that caspase-8 plays a non-apoptotic role in the cell, such as regulating cell differentiation and senescence [29]. We assumed that the activation of caspase-8 observed after YLT322 treatment is a possible downstream "bystander" event which has been reported elsewhere [30]. These data suggest that the intrinsic apoptosis pathway is more important than the extrinsic pathway in apoptosis induced by YLT322.
A key step in the intrinsic apoptotic pathway is the disruption of the mitochondrial membrane, which triggers the release of cytochrome c from mitochondria to cytosol. From our study, a reduction in ΔΨ and a release of cytochrome c from the mitochondria into the cytosol were observed in a dose-dependent manner after YLT322 treatment.
Bcl-2 protein family plays an important role in the regulation of mitochondrial apoptosis pathway [7]. Multidomain pro-apoptotic proteins such as Bax and Bid can open the ion channel in the outer mitochondrial membrane in the presence of a death signal, leading to mitochondrial membrane permeabilization and release of cytochrome c into the cytosol [31], [32]. Cytochrome c in the cytosol binds to the scaffolding protein Apaf-1 and pro-caspase-9 to form the apoptosome, which triggers the apoptotic cascade reaction thereafter. The anti-apoptotic Bcl-2 family members such as Bcl-2 and Bcl-xl inhibit cytochrome c release by blocking the activation of pro-apoptotic proteins. For most cancers, the over-expression of Bcl-2 protein associates with poor survival, uncontrolled progression and resistance to anticancer agents, making Bcl-2 family members promising anticancer drug targets [24], [33]–[34]. Our results showed that the expression of Bcl-2 decreased in cells treated with YLT322, whereas that of Bax increased. We assumed that cytochrome c release from the mitochondria into the cytosol upon YLT322 treatment is due to the down-regulation of Bcl-2 and/or up-regulation of Bax.
Studies have suggested that Akt/p44/42 MAPK pathway plays essential roles in apoptosis and could be modulated for cancer prevention and treatment [35]–[37]. Here, we found that treatment with YLT322 decreased the phosphorylation of both Akt and p44/42 MAPK. This suggests that Akt and p44/42 MAPK may contribute to the apoptosis induced by YLT322.
YLT322 treatment on nude mice bearing tumors resulted in significant inhibition of tumor volume without any apparent sign of toxicity. Immunohistochemical studies revealed that the regression of tumor size in mice models by YLT322 was also associated with the activation of apoptosis, as demonstrated by of the presence of active caspase-3-positive and TUNEL-positive cells in tumor xenograft samples. YLT322 also caused a decrease in Ki67-positive cells, suggesting a reduction in cell proliferation.
In conclusion, our study demonstrated that YLT322 inhibits cell growth/proliferation by inducing apoptosis via the intrinsic pathway. In addition, YLT322 suppressed the growth of human liver cancer xenografts without significant toxicity, suggesting that it may be a potential candidate for cancer therapy. In depth studies are still needed to elucidate the precise targets mediating the anti-cancer effects of YLT322.
Author Contributions
Conceived and designed the experiments: YLT SXJ XY. Performed the experiments: SXJ XY YTH. Analyzed the data: YLT SXJ WNY ZYX. Contributed reagents/materials/analysis tools: XYZ ZLD SXH SYJ. Wrote the paper: SXJ XY WNY.
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- 14. Ali S, Banerjee S, Ahmad A, El-Rayes BF, Philip PA, et al. (2008) Apoptosis-inducing effect of erlotinib is potentiated by 3,3 -diindolylmethane in vitro and in vivo using an orthotopic model of pancreatic cancer. Mol Cancer Ther 7: 1708–19.
- 15. Xu YZ, Zheng RL, Zhou Y, Peng F, Lin HJ, et al. (2011) Small molecular anticancer agent SKLB703 induces apoptosis in human hepatocellular carcinoma cells via the mitochondrial apoptotic pathway in vitro and inhibits tumor growth in vivo. Cancer Lett 313: 44–53.
- 16. Cory S, Huang DC, Adams JM (2003) The Bcl-2 family: roles in cell survival and oncogenesis. Oncogene 22: 8590–8607.
- 17. Ratajczak J, Majka M, Kijowski J, Baj M, Pan ZK, et al. (2001) Biological significance of MAPK, AKT and JAK-STAT protein activation by various erythropoietic factors in normal human early erythroid cells. Br J Haematol 115: 195–204.
- 18. Kline MP, Rajkumar SV, Timm MM, Kimlinger TK, Haug JL, et al. (2008) R-(-)-gossypol (AT-101) activates programmed cell death in multiple myeloma cells. Exp Hematol 36: 568–576.
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Champions held in season opener
Posted: 7 August 2009 Time: 22:29
Defending Absa Premiership champions SuperSport United were held to a 1-1 draw at Maritzburg United in the 2009/10 season opener on Friday night.
Morgan Gould had given the Tshwane team the lead at Woodburn Stadium, but Matsatsantsa were then outdone by their former player Noah Chivuta, who converted a penalty late in the first half.
However, SuperSport coach will be more concerned by the health of new signing Petrus Mahlatsi then the dropped points. The striker lasted just three minutes of his debut before being rushed to hospital after a clash of heads with Byron Hendricks.
SuperSport certainly had the better of the first half, looking the more threatening and cohesive side on attack, and also looking more assured in defence.
The visiting side had the first chance of the game in the fourth minute when Glen Salmon turned sharply in the box and beat keeper Brendan Wardle at the near post, only to see the ball strike the upright and bounce back into play.
Maritzburg struggled to find their rhythm throughout the first half, but came within a whisker of scoring in the 18th minute when Man of the Match Musa Nyatama's glancing header from a corner-kick just missed finding the far post.
Six minutes later Wardle was again found wanting as Klate swung in a dangerous free-kick and Gould rose well to nod the ball into the unguarded net.
SuperSport should have been 2-0 ahead in the 26th minute when Nkosi delivered a free-kick into the box, but an unmarked Bongani Khumalo got his header all wrong.
Maritzburg had hardly troubled the United defence, but in the 40th minute striker Brice Aka did well to draw Gould into a foul just inside the box. The penalty was awarded and Chivuta held his nerve to pick out the bottom right-hand corner.
Maritzburg got the second half off to a flying start and could have gone ahead just seconds after the restart when Peter Petersen's long throw in reached Obada, but the Nigerian miscued his close-range effort.
SuperSport put together a great team move in the 51st minute which resulted in the ball falling kindly for Hlompho Kekana, but he crashed the ball over the bar from 12-yards out.
More efforts then followed from Gordon Igesund's side as Rudolf Bester fired straight at Dennis Onyango and Vito Badiane had a thunderous effort blocked by Gould.
The momentum stayed with Maritzburg for the remainder of the match as they created several more chances through the likes of Felix Obada, Chivuta and Kulelani Madondo, but none which troubled Onyango.
HOME: Maritzburg United
- 1 brenden wardle
- 13 Byrone Hendricks
- 24 james madidilani
- 24 Peter Petersen
- 30 dikilu bageta
- 4 Musa Nyatama
- 7 Felix Obada
- 21 Noah Chivuta
- 25 Rudolf Bester
90 mins - Diego Brown
- 26 Vito Badiane
67 mins - Kulegani Madondo
- 27 Brice Aka
56 mins - Fadlu Davids
AWAY: SuperSport United
- 36 Denis Onyango
- 4 Bongani Khumalo
- 4 Tsietsi Mahoa
- 29 Morgan Gould
- 34 Rudi Isaacs
- 7 Daine Klate
- 8 Hlompho Kekana
- 8 Jabu Maluleke
69 mins - Petrus Mahlatsi
80 mins - rudzani ramudzuli
- 10 Siyabonga Nkosi
90 mins - Letladi Madubanya
- 9 Glen Salmon
- 28 Anthony Laffor

subset: commoncrawl


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Time to plant the Amaryllis bulbs
My son's mother-in-law gave me a beautiful Apple Blossom Amaryllis bulb for Christmas. To be honest, I've been putting off writing about it because I wasn't sure what the correct term was to refer to her. Come to find out, there is a good reason for my confusion. There is no term for your child's in-laws. They may be referred to as your child's in-laws or as your (in my case) daughter-in-law's parents. Some circles recommend co in-laws but that just sounds silly to me and besides it is impossible to write - is that co-in-laws, co in-laws, co in laws or what? But I digress...
I planted the amaryllis bulb on December 28. According to the experts, the flower should bloom in 6 to 8 weeks, but some sources predict as many as 12 weeks before blooming. I'm guessing my Amaryllis will bloom sometime in mid to late February, but I'll keep you posted on its progress.
To learn more about Amaryllis flowers and to view the many varieties, please visit Amaryllis Care: Bold, Exotic Flowers for the Holidays.
Until next time .... HAPPY GARDENING!
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May 2018


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Kim Kardashian has congratulated her boyfriend Kanye West after his success at the Grammys.
West picked up a total of three gongs at the ceremony, winning for 'Best Rap Performance', 'Best Rap/Sung Collaboration' and 'Best Rap Song' with The Throne collaborator Jay-Z.
Kardashian - who is currently staying in Brazil with West - took to her Twitter page this morning to congratulate her beau on his victory.
"Awww just waking up in Brazil, seeing Kanye won 3 Grammy's last night!!!" she wrote. "That makes a total of 21!!!! That's just so insane! Soooo proud!!!"
Kardashian had revealed last night that she was keeping track of the Grammys online, writing: "In the car heading to carnival here in Rio! Good thing there's all this traffic so you can look up all these hot Grammy looks!"
She singled out Jennifer Lopez - and her controversial leg bombing - for particular praise, saying: "Wow @JLo looks the best hands down!!!!! So sexy!!!"
West and Kardashian were photographed sightseeing in Rio de Janeiro over the weekend, visiting the famous Christ the Redeemer statue and walking around with Will Smith.
They are currently expecting their first child together, although Kardashian is said to be suffering from pains related to the stress of her divorce proceedings with ex-husband Kris Humphries.
Photo gallery - Grammy Awards 2013 winners:Music: Grammy Awards 2013: Winners


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(The following is a slightly modified version of a comment left in response to a blog post titled "An Open Letter From The Patriarchy". Upon posting it I realized that their comments are moderated and, not wanting to leave it to chance, decided to publish it here as well.)
Entertaining. I would categorize this as "high fantasy". Whoever penned this is incredibly creative. I give it a 10/10!
…I just hope that, for their own sake, no man takes this seriously.
"[T]he greatest trick the devil pulled was convincing the world that he didn't exist." A conveniently quoted old saying, because The Devil, in this instance, is woman and her power. This is something almost all women know. I believe the only women oblivious to their own power are Feminists, and even then, it's only a very small fraction of their ranks. These plays at victimhood are indirect power-grabs, and men project uncharacteristic frankness onto women in believing that any meaningful number of them actually see themselves as victims.
I think women's power is something all men recognize, at least subconsciously, and the classically boisterous masculine ego is but a defense mechanism ("Reaction Formation") arising in response to that uncomfortable truth. Such a strong response betrays the male ego's attempt to deny what it already knows…
But…do not take the above as an assertion of submission. On the contrary, simply stating what I have above is, on its own, a challenge to woman's power, and if you cannot understand why, then you have a lot to learn…
– Jack The Abandoner


text:
Content: See & Do
Category: Accommodation
Old Village Post House Inn
101 Pitt ST
Mount Pleasant, SC 29464Get Directions
Phone: (843) 388-8935
City: Mount Pleasant
Region: Coastal & Islands
Old Village Post House Inn
In addition to tasty fare and gracious hospitality, the Old Village Post House Inn offers historic and comfortable accommodations for overnight guests. Six distinct and well-appointed rooms provide a choice of sizes and style. Amenities include wireless high-speed Internet, cable TV, telephone, private bath with whirlpool tub, daily breakfast and more. Set in the quaint, historic fishing community of the Old Village of Mount Pleasant, the Inn makes its home among centuries old oaks and harbor-side homes. It's just minutes from downtown Charleston and the beaches on Sullivan's Island and the Isle of Palms.


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Low Tox Living Workshop
Sat., 12 May 2018, 10:00 am – 11:30 am AEST
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Wagga Wagga, NSW 2650
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A introduction to reducing toxins within your home and body. In this workshop I will teach you the benefits of using essential oils, DIY and low tox products to help you focus on the areas you are seeking to improve.
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My system is not allowing me to capture footage. It usually states that the device is offline. At times it will show and allow me to bring in footage, although this is extremely irratic. I have never been able to be able to control the playback device through premiere, or any other program. My computer, however, does read the playback unit. I am trying to use my XL2 through firewire to bring in the footage. What am I doing wrong?
More information needed for someone to help... click these links and provided the requested information
Thanks so much for the help. I didn't corralte the issue to be with Windows 7. Up and running and very happy!


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Danon jewellery: Bangles
We love this simple bangle with its rustic hammered finish. The sturdy bangle is complimented by two small heart charms- one smooth and gold, the other hammered and silver.
Heart Charms: approximately 11mm
Bangle Diameter: 6.5cm
Packed in a giftbox as standard. We offer free UK delivery on all standard orders!
Danon jewellery is made of sturdy pewter and glazed with silver and/or gold for optimum lustre. Danon jewellery fuses classic and contemporary styles for immensly wearable pieces of jewellery that you will love.
Additional Information
|* Simple Danon Jewellery: Bangle with Contrasting Mixed Metal Heart Charms
|Danon designer Jewellery


text:
Introducing myself
Hi everyone! My name is Terri and my passion is drag racing. I have been involved in this sport for 17 years now and my family have always supported me in this motorsport and...
Started by Terri on 07/24/2010 in Mums Who Are Involved In Drag Racing At Race Tracks
Last update on 03/17/2015 by Lucitta


text:
Ballweg Custom Sawing
6498 County Road KP
Mazomanie, WI 53560
1-608-643-6869 (fax)
Call ahead - Most orders are ready in less than a week, may be done in 24hrs.
Family owned business in the Sauk Prairie area who provides custom sawing. Rough cut lumber 4' - 20' lengths, your trees or ours. Air dried, ready for your project. We specialize in dump truck side-boards & low-boy planking. Firewood & horse fencing lumber are also available.
From Sauk City, travel Hwy 12 East for approximately 5 miles; turn right onto County Hwy KP and then the mill is on the left hand side.


text:
Kolkata, Feb 2 :
Former West Bengal Chief Minister and CPI-M leader Buddhadeb Bhattacharjee Sunday admitted that the Netai incident, in which nine people died, was a "grave mistake" committed by the party workers.
Nine people, including four women, were killed and 28 others were injured in the January 2011 clashes, triggered by assailants allegedly backed by the Communist Party of India-Marxist (CPI-M) in Netai near Lalgarh of West Midnapore district.
Reacting to the development, the state's ruling Trinamool Congress demanded the immediate arrest of Bhattacharjee for his party's complicity in the killings.
"The Netai incident was a mistake — our lads committed a mistake, a big mistake in Netai," Bhattacharjee said while addressing a party rally in Midnapore of the district.
Bhattacharjee, who earlier had regretted the incident, however, accused the Trinamool Congress of unrepentantly carrying out atrocities on innocent people and rival party workers.
"We have committed a mistake, but what about them (Trinamool)? They have been carrying out atrocities all across the state. They have never regretted doing such acts," he said.
Reacting to Bhattacharjee's admission, Trinamool Congress leader and state Municipal Affairs Minister Firhad Hakim demanded immediate arrest of the CPI-M politburo member.
"When he has admitted his own men carried out the carnage, then he must be immediately arrested and tried before a court of law. Instead of trying to apologise, he should surrender himself immediately before the police," said Hakim.
Trinamool secretary general and Parliamentary Affairs Minister Partha Chatterjee ridiculed Bhattacharjee for his "apology".
"Considering the number of times he has apologised for the mistakes committed during his regime, he should be the Olympic champion for apologies. But people have realised and such apologies will not have any effect on them," said Chatterjee.


text:
It's hard to sum up the last few weeks in a single post, so I won't try. What I can say at the moment is that the trip was amazing and spending time with family and friends over the holidays was simply the best gift we could have received.
Even better was watching Miss K interact with family. She was so happy to have them around and let's face it, all that extra attention and affection.What 3 year old wouldn't soak that up?
I made the decision to not bring my dSLR camera with me so I would actually relax and not spend the whole time behind the lens. We did bring a small digital camera with us but it blurred so many images inside we didn't end up using it much. In a way, I'm not upset in the least as we really did participate in the trip rather than me watching it from a distance.
All in all, it was a very relaxing trip. We spent time in two cities where our families are, and one day in between in Vancouver, where we used to live. We had a couple large get-togethers so did most of our visiting with people then and therefore didn't spend the entire time rushing around trying to visit. It was such a change from how it often was at the holidays.
The flights were... interesting. It actually went very smoothly, but three different planes, a long 9 hr flight over the Atlantic and over 24 hours of travelling each way was exhausting. Miss K did pretty well with it, all told, except for that slight problem where she had discovered how to do up and undo her seatbelt buckle and refusing to leave it on during take off and landing. When we insisted, she'd erupt in a huge tantrum and hubby would have to lean over and hold her with his hand over the buckle until we reach elevation and the seat belt light went off.
Fun times.
One of the biggest changes in recent weeks has been her vocabulary. She is repeating words like crazy and it's amazing how many she comes up with! Furthermore, we are noticing her trying compound words (like pineapple) and using more than one word (apple juice). No real sentences yet, but the improvement inspires hope that she will need little intervention in the end.
There's much more to say, but for now I wanted to say that I'm back and catching up!
Happy New Year. I hope this year brings wonderful things for you all!
Wednesday, January 9, 2013
Where I Poke My Head in and Say 'I'm Back'!
3 comments:
I'm exhausted, lol...but it is a good exhausted and loved every minute we got to spend with all of you!ReplyDelete
So glad there's been improvement with her vocab! That's great!ReplyDelete
I'm glad you got to spend the holidays with family :) I'm sure it was chaos, but the good kind.
Happy New Year! I'm so glad you had such a nice trip and that is amazing news about Miss K's vocabulary. What a wonderful way to start the New Year!ReplyDelete
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Plus... there is a blog hop over at the Art Impressions' blog, here is the link!
And here is the order!
Thanks so much for stopping by and happy hopping!
Perfect paper for these gals, Kerry! Great card :o)
Oh I love the bright colors you chose. And the patterned paper is really cool too.
Love the coloring of the images and the speech bubbles in the background.
Love this image and coloring. I am new to art Impressions line and will definitely be ordering the BFF stamps.
So CUTE!!! Love it, Kerry!
hugs, Sharon
The background paper is the bomb! It was perfect for this image. Great coloring, I am jealous.
Love it and love these new stamps!
Love the colors! Very cute card :)
Really fun design and fabulous coloring, Kerry! The bg paper is perfect.
Great card and I love the new stamp
Looks marvelous! Love the color choices
Aack! I love your coloring -- what a fabulous job, Kerry! I can't wait to get this set. Thanks for sharing!
Fun card! Love your coloring and the simple layout which allows the focus to stay on the image.
Great coloring. Love the layout too.
Thanks for sharing.
Reminds me of my daughter and her friends sitting on the couch texting each other...lol. :)
Great card, love these girls !
Corrie x
Great card, love these girls !
Corrie x
Love this image and coloring! Very cute and fun card! :)
Love these bright colors!! This is funny card =))
Great job. Love the color combos and choice of paper.. it works perfectly.
Love the colors. Great card
Kerry, your background paper is so perfect for the image. Love this!
Love these new AI stamps. Your card is super - love the bright colors and design.
Gotta love the AI girl friends!
This is so adorable, I love it.
It is pretty cool! Love it! :)
This is such a fun set! Great job on your card.
cute set love the different ways i've seen the same set
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Love the girlfriends and you are a master colorer (is this a word?! well you know what I mean).
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I'm loving these new BFF sets! Need one in every color, know what I mean? Ha! :)
Brilliant card love your work
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Love your card Kerry!!! Reminds me of the kids texting each other while sitting in the car!
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text: expected to average 85 million barrels per day in 2006, leaving just 1.9 million barrels per day of excess production capacity that could be called upon in an emergency, according to Cambridge Energy Research Associates. CERA oil analyst James Burkhard said this supplycushion shows no sign of growing anytime soon. "So far we're not at the point where demand is pulling back," he said. However, global oil demand is growing at a slower rate than last year, according to the International Energy Agency.


text:
Archive for April, 2008
Anza Borrego Desert State Park
Font's Point, Anza Borrego Desert State Park, San Diego County, California
Perched on top of the cliff at Font's Point overlooking the twisted badlands that runs as far as my eyes can see is one of the many wonderful gifts of nature that makes me almost in tears from awe and from amazement.
Font's Point, Anza Borrego Desert State Park, CA
" Four milion years of geologic and paleontologic history are exhibited across the stark desert landscape here at Font's Point. This may be the best place in North America to view sediments of the Pliocene and Pleistocene Epochs." – http://www.desertusa.com/anza_borrego/du-abpbobad.html
Being up close to the magnificent power of nature, hard it may be to fathom, always brings me a positive feeling of gratitude and a yearning for understanding, so we all can live harmoniously in this beautiful but fragile world.
Multi-Hued Borrego Badlands, Anza Borrego Desert State Park
The desert, may look harsh and unforgiving and hosts one of the most fragile ecosystems on our planet, yet its sculpted formations is also one of the most beautiful places on Earth. Its unique sense of peacefulness, its exposed twisted layers of the earth so vast to fathom, and its surreal beauty are all there for us to look deeper to hopefully learn and understand more. Barren it may be, yet the desert ladscape becomes a paradise of colors in early spring after a wet winter.
Desert Valley turns into a Paradise of Color, Anza Borrego Desert State Park
The desert sand verbena and evening dune primrose brings a natural fragrance into the air.
Fragrance that fills the whole desert valley along Henderson Canyon.
Wildflowers at the end of Di Giorgio Road
However, this display of wildflowers leaves as soon as it comes. The wildflower at the valley floor usually peaks first and lasts for about a month. After the valley floor, the higher elevation and the cacti follows the bloom.
Brittlebush, Anza Borrego Desert State Park
Blooming Ocotillo, Anza Borrego Desert State Park
Blooming Beavertail Cactus, Anza Borrego Desert State Park
Blooming Cacti, Anza Borrego Desert State Park
If you come to Anza Borrego to see this other desert located at the eastern side of San Diego County, do make a stop at their Visitor Center ( http://www.desertusa.com/anza_borrego/visitor_new.html ), which is located 2 miles West of downtown Borrego Springs. They have very interesting exhibits and videos about the geological forces that shaped the landscape, life forms and human stories of the Anza Borrego region, and features a small slot canyon entrance. The video will not only whet your appetite to see more of Anza Borrego at different seasons, but helps you learn more about the ecosystem of the park and gives an idea on interesting places to go. The park rangers are very helpful also in answering questions about the features of the park. Outside the visitor center is a nature trail that gives an overview of the plant habitat around the park.
For detailed hiking information and guide, we usually use Jerry Schad's " A Foot and a Field in San Diego County." However, I do not see an on-line link to this wonderful hiking book, thus for some on-line informatioon about the hiking trails in Anza Borrego, please visit this site: http://www.desertusa.com/Thingstodo/du_absp_hike.html. A map of Anza Borrego Desert State Park is also provided in that link.
For more description of Anza Borrego Desert State Park and its Natural History, please visit this link: http://www.desertusa.com/anza_borrego/du_abp_desc.html
If you get a chance to visit San Diego, and have some interest in geology and the natural wonders in a desert landscape, do make some plans to visit Anza Borrego, and experience the unique plant and life habitat
The Cycle of Life, Anza Borrego Desert State Park
that exists in the midst of the rocky geography and corrugated badlands.
Green Bushes Amidst the Multi Hued Badlands, Anza Borrego Desert State Park
Come, and be amazed with the wonders of Anza Borrego, The Other Desert. And the other side of the more known coastal landscape of San Diego County, California.


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Brown Salad Plate
Details https://www.belk.com/p/herend-brown-salad-plate/7601194ABGNM101518000.html
Additional Information
Princess Victoria Dinnerware is the perfect name for a pattern that incorporates a well-known motif from Queen Victoria but is not trying to invoke the same presence as the Queen herself! The perpetual elegance of this prominent plate will add a touch of sophistication to any family meal.
- 7.5-in. diameter
- Gold banded
- Hand-painted finish
- Porcelain
- Hand wash
- Imported
This item is part of the collection: Princess Victoria Dinnerware - Brown
Shipping Details
Shipping Details
This item ships directly from the vendor, usually within 4 business days. Shipping Details
Due to shipping weight restrictions, this item cannot be shipped to a PO Box.
Return Details
Returns are easy at belk.com. Easy returns


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NASHVILLE (AP) — Willie Nelson said Thursday that he will turn an upcoming Texas concert into a benefit for victims of the explosion at a fertilizer plant not far from where he grew up.
The country music icon still has a home in Abbott, Texas, about five miles north of West, which was rocked by the explosion Wednesday night that left an unknown number of people dead and more than 160 hurt. He remembers riding his bike the short distance between the towns and still has many friends and family there.
"Our hearts and prayers go out to the people of West," Nelson said in an interview before taping a CMT Crossroads special in Nashville. "There are a lot of our friends and loved ones and neighbors down there. We talked to some of them and some of them made it out OK, and some of them didn't. But they're strong and they'll be back. It's one of those things you don't get over. But you will get through it."
The concert is scheduled for April 28 in Austin.
When not on the road touring, Nelson has lived in Austin since 1971. The concert was scheduled as an 80th birthday celebration, but now will serve a dual purpose.
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Pocket projectors hit the streets just a few months ago, and have already created a buzz in the tech world. From the the incredible creative potential of the DLP pico projector kit we discussed a couple weeks ago or the LCoS version we tweeted a few days ago – a large quantity of digital bits have been dedicated to the products on tech blogs.
But what has yet been left out of the conversation is the answer to the question "How do the LED pocket projectors compare to slightly larger – but similarly priced – LCD and DLP variety of portable projector?"
So, today we are whipping out our demo pocket projectors from 3M and Optoma and throwing in a sub-$600 portable projector for some comparing.
3M MPro 110 Pocket Projector
The 3M MPro 110 projector is one of the few new pico products manufactured by a brand you've heard of. There are a few phone makers that have integrated the pico technology into their cell phones, but for those dedicated to projection alone, Optoma and 3M look to be two of the major players. Although I did just stumble across a lesser known brand that has on board content like MP3 files, etc. which is a nice idea. I have not heard of the company however, and without the telephone function, who knows if it will catch on.
Ok, back to the 3M MPro 110. What's cool about this unit, other than it's tiny size and nice looking LCoS iamge, is the VGA connection. The VGA functionality makes it more friendly for use with a laptop, and with the additional input for composite video, it also means connectivity flexibility with some other portable video sources.
The 3M also has a slight offset, which means the image will project below the direct line of the projector lens. So, if you shoot your projector at a screen, your image will show up slightly below the line of the lens, rather than straight ahead as you might expect. This might be useful in some situations, and inconvenient in others. It probably won't make a difference to most users.
3M MPro 110 pocket projector in action with a small rear projection screen.
Video found on YouTube.
Optoma PK-101 Pico Projector
We have already shown some video and posted news about the Optoma PK-101 projector, and by my non-scientific tally, this unit has received the most press from the category. There is some good reason for that. It's initial target users was the iPhone/iPod user, and was first released in Japanese Apple stores. While the PK-101 lacks a VGA connection like the 3M model, it does have a small (1 watt) speaker on the unit. We have our on-the-fly video of the PK101 in action on a plane. (see video)
Video : Pocket Projectors versus a Cheap LCD Projector
Okay, now that we have discussed our two pocket projector players, let's get on to our video comparison between the small wonders and their big daddies.
Note : I should mention that our demonstration is a challenge for a video camera, even a good one like the Panasonic DVX100B profesional camera we used. So, in person, the pocket projectors actually looked a little brighter, particularly after your eyes adjusted to the room. However, the dramatic difference in brightness is captured well in the video.
If you watched the video, what you saw was not entirely fair. It's a little like a big brother stuffing his little sister in the dryer and turning it on for a couple minutes – at least that's the memory it returned for me. However, some day soon, little sis is going to grow up and get her revenge. While the pocket projectors may not be the brightest now, the next generation of pocket projectors will no doubt be bigger, stronger, prettier, and better.
So, if you like being the guy or gal with the coolest tech gadgets, the one that had the first home theater on the block, or the home automation system, you might consider getting your hands on one of these projectors ASAP. They are in limited supply, and have a pack a nice WoW factor.
If you'd rather wait for the pocket projectors to become the standard for portable projection, and even home theater, you might not have to wait long. With energy efficency, lamp life over 20,000 hours, battery opperation, and their size, they have too much going for them not to continue to advance the technology. Here is a quick take on some advantages of LED based pocket projectors like the Optoma PK-101 and the 3M MPro110:
Advantages of Pocket Projectors :
- Very small. Can be slipped into a pocket.
- Very light weight at just a few ounces
- Wider color space & good color saturation
- No "rainbow" on DLP variety (LCos does not have rainbows)
- 20,000 – 30,000 hour lamp life
- Battery opperated – no power cords required for presentations under an hour
- Turns on in seconds. Instant turn off with no cooling time required
- Environmentally friendly with LED lamp's low power consumption
- LED lamps are 100% mercury free
Disadvantages of Pocket Projectors :
- Not bright enough for medium or large groups
- Must be close to screen/wall surface
- Fewer connectivity options
- Serious audio limitations
- Low resolution
Is It Pico or Pocket?
We have also been asked if the proper name for these tiny beamers is "pocket" or "pico." It seems the industry is yet to standardize on one term, but this blogger believes "pocket" will eventually take the prize.
Why? Because pico projector is an industry term that has been used to describe projectors under four or five pounds, AND, pocket gives the end user an immediate sense of exactly how small these projectors are. There are some 'pico' projectors that would make for a very bulgy pocket. So if you're shopping for a pocket projector, don't be confused by the pico's already on the market. No disrespect to some very good, very small pico projectors (like the Mitsubishi PK20), but the pocket projectors are indeed small enough to slip into your pocket. Of course, I could be wrong. Maybe they will ultimately be called 'micro' projectors, or 'bitty beamers', or 'video pointers', or 'petit projectors'…


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Ottawa Kijiji is a free classifieds site for residents of Ottawa, Canada. The online site is located at Kijiji.ca. The company offers free classified sites for various metro areas in Canada including Toronto, Vancouver, Montreal and Calgary.
The classified ads at the site include job postings, used vehicles, available pets, real estate and community groups. The site also features personal ads, ads for services needed and a link for swap and trade customers. Customers have the option of registering for a user account on the site, browsing categories, searching by keyword or changing their location. The site also allows customers to post their own classified ads.


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Today I want to talk about God. And how amazing He is. I've always believed that everything happens for a reason. The reason may not be immediately apparent but eventually the purpose will be revealed. And we will understand what used to be unclear. I want to share a story. G's story. I may have already written about him in previous posts but let me still give a short background.
I met G when I was in grade school, 3rd grade to be exact. Young as I was, I already developed a crush on him. This carried on until we were in 6th grade and that was when I was able to convince myself I loved him. I loved him up until high school. Even when I was in college. We went through a lot of admitting our feelings, almost getting together moments. But it never worked out. There was always something in the way. But all those years, I've prayed to God. I prayed that God would give us a chance to be together. He was the love of my life. I didn't see myself being with anyone else. We lose touch, then we rediscover our connection. It was a never ending cycle. Everyone thought I was wasting my time and energy with a love that was never meant to be. But still I prayed to God for him. I thought if I persevered enough and God sees how much I wanted it, He will eventually hear me.
In 2008, after leaving my first job, I thought God has finally heeded my prayers. G sought me out and everything fell into place. I thought all those years of waiting have finally paid off. I thought I hit the jackpot. Winning the lotto would not have compared to how happy I was. Not long after, things started to go sour. I began to see how different we were. The problems started to pour in. I was hurting. And I was disappointed. But because I've prayed so hard for it, I wanted to try a little harder to make it all work out. But in the end, my little bubble still burst. I kept asking God why. I've waited so long and it ended so fast. I really thought God hated me. My one wish, my one ticket to be happy, He really had to take it away.
I eventually stopped crying. And that's when I realized that God didn't hate me. God loved me so much and that's why He took G away. Because G and I do not make a great team. Because G hurt me. It's not the life God wanted for me. No matter how much I wanted G, God took him away because we wants me to have someone better. He gave me a glimpse of what life will be like with G so I will realize how wrong he was for me. God wanted me to have closure. He wanted me to stop pining for him. He wanted me to move on and find a better partner. God knows what's best. We only need to have faith in His plans and His timing. Our darkest time will eventually reveal a silver lining. I am happy now. I like how my life turned out after I closed the G chapter. Life would have been so different had we ended up together.
And that's why I believe everything happens for a reason. It may take time before we realize it, but it's there. We just gotta believe it.


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The iconic graffiti artist KATSU's Mark Zuckerberg posters have been put up within New York's lower Manhattan, Chelsea and Williamsburg neighborhoods. The social media icon is featured with a bruised eye and cracked teeth, a far cry from the pristine way in which he is most often depicted.
Titled Status Profile, KATSU aims to address the release of personal data through Facebook as well as the coming future. Given the beaten up nature of the posters, the artist has made the case for rife opposition towards Facebook and akin forums.
"With the internet being literally woven into the human experience I cannot stand by and not voice my discontent with those who plan on owning our lives," said KATSU via e-mail.
Pulverized Street Art Posters
More Stats +/-
Graffiti-Clad Workspaces
Persistent Narrative Graffiti
Vibrantly Multi-Layered Graffiti
Guerilla Supermarket Art
Astoundingly Surreal Street Art
The KATSU Mark Zuckerberg Images Display His Face in New York
- By: Jordy E Apr 4, 2013
References: eyebeam.org & gizmodo
References: eyebeam.org & gizmodo


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in his mother's new house …
the clip-clop
of his Wild Horse skateboard
fading into the night
Favorite Tanka, 2016 Tanka Society of America Members' Anthology
Chen-ou Liu
Selection & Reflections by Beverley George (posted on TSA Facebook page, July 20): This compelling tanka creates a mood; tells a story; leaves the reader conjecturing. So much depends on the simple word 'new'. It raises the question of why the son has no access to his mother's home; hints that he may have had to the former one. The brand-named skateboard suggests that he is young.The first and last lines bookend the poem with darkness. Sound plays its part with 'clip-clop' linking to the skateboard's name and 'fading' adding to the feeling of separation and loneliness. A short story in five lines.


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Autism Speaks, a leading autism advocacy organization, dedicated to advocating for the needs of individuals with autism and their families, is offering $5,000 grants to local service providers addressing the areas of education/training, recreation and community activities, young adult / adult services and technology. Organizations can apply for grants on the Autism Speaks website: https://www.grantinterface.com/autismspeaksfs/Common/LogOn.aspx.
This is a great opportunity for local businesses. For more information on what your organziation can do to get a grant, visit the Autism Speaks Local Grants page: https://www.autismspeaks.org/family-services/grants/local.


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Gardening, Containers, Planning, Garden Coaching, Perennials, Annuals, Shrubs, Evergreens, Bulbs
Tuesday, February 14, 2012
Not Your Old Fashioned Petunia
16 comments:
- Bernie H said...
So many brilliant colours! I just adore 'White Russian' and 'Hell's Bells' would be brilliant out in my courtyard. I hope some of these make their way to my corner of the world this year.
- Tue Feb 14, 06:31:00 AM 2012
- Gatsbys Gardens said...
Bernie, I think I read that a couple of these new introductions were originally from Austrailia.
- Tue Feb 14, 08:07:00 AM 2012
- Larry said...
We were so amazed at out Vista Bubblegums this past season that we have ordered the three larger vistas in three colors for the coming season. The problem will be finding space for them as they do so very well... Larry
- Tue Feb 14, 08:12:00 AM 2012
- Lona said...
Such wonderful varieties are showing up with such wonderful colors. I do not think the black ones will last either. I have a hard them getting them to fit in with other flowers for some reason. I am one of those gardeners that love petunias in the garden even when I have to deadhead them.
- Tue Feb 14, 09:42:00 AM 2012
- Gatsbys Gardens said...
I will have to look up those larger vistas. I can't imagine any better than Bubblegum!
- Wed Feb 15, 04:36:00 AM 2012
- Gatsbys Gardens said...
I like petunias also and don't mind cutting them back to get more blooms.
- Wed Feb 15, 04:37:00 AM 2012
- Beth said...
Beautiful and springy! We too liked the Vista Bubblegum last year. I hope you had a nice Valentine's Day.
- Wed Feb 15, 12:16:00 PM 2012
- Zoey said...
I like petunias, but am one of those who don't like the maintenance required to keep them looking good. Even the ones you don't have to deadhead, still look ugly until I cut them all off. . . hate the sticky fingers, too.
- Wed Feb 15, 02:42:00 PM 2012
- garden girl said...
I'm not a big fan of petunias, but then again, if I had more sun I'd probably give some of the newer varieties a try. I do like the colors, and I love the black ones.
- Thu Feb 16, 07:35:00 AM 2012
- Gatsbys Gardens said...
Thanks Beth,
According to Larry there are other vistas also so I will look into this.
- Thu Feb 16, 07:40:00 AM 2012
- Gatsbys Gardens said...
Hi Zoey,
I just cut them back around July with the clippers, feed them and avoid the sticky fingers.
- Thu Feb 16, 07:41:00 AM 2012
- Gatsbys Gardens said...
Hi GG,
I know there are many people who do not like petunias but for some hot locations they are great performers.
- Thu Feb 16, 07:42:00 AM 2012
- linniew said...
Wow what a lot of colors! I think the trick with petunias is to mix them in with other plants. They can trail around, get tall, lean-- they seem to keep flowering no matter what and fill in around perennials.
- Thu Feb 16, 08:48:00 PM 2012
- CanadianGardenJoy said...
Eileen girl I still love petunias and these new types are beautiful !
Some gardeners turn their noses up to them .. but I have a weak spot for them .. the blue/purple veined ones are so gorgeous .. and of course the black mixed with orange ? perfect Halloween combo ! LOL
- Fri Feb 17, 12:26:00 PM 2012
- Pat@Back Porch Musings said...
Beautiful petunia images, Eileen.
Thanks so much for visiting the Back Porch, during J's hospitalization. We both appreciate it.
- Sun Feb 19, 07:39:00 AM 2012
- Becca's Dirt said...
Those are some beauties. Love all the many colors.
- Tue Feb 21, 11:00:00 AM 2012


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Here are some of the top landscape designer schools which we are delighted to share with all,
1. The Anna Gresham School of Landscape Design
This famous school has been teaching landscape design worldwide for over nine years. They have established the global reputation of being the top in the field of landscape design education. They are the landscape designers ourselves and training world class landscape designers is their dedicated mission.
Their comprehensive study program of theory and practice covers every aspect of landscape and garden design. By working through many real design assignments, one will learn and practice all the necessary skills of a landscape designer.
2. Penn Foster Career School
There are certain skills you need to begin a career as a Professional Landscaper. The Penn Foster Career School Professional Landscaper Program helps you to learn quickly and conveniently.
You'll learn about:
Plant health, soil, and fertilizers
Servicing garden tractors and other tools used in landscaping
Creating landscaping designs using shrubs and ground covers
How to start operating your own landscaping business
Servicing garden tractors and other tools used in landscaping
Creating landscaping designs using shrubs and ground covers
How to start operating your own landscaping business
3. Kingston University London
- Architecture at Kingston University highly focused on the design, reflecting the course's context within an art and design-based faculty. The curriculum builds on the fundamental creative processes of observation and making, and its ethos could be summarized as "thinking through making". Completion of this degree automatically gives RIBA (Royal Institute of British Architects) Part 1 exemption priviledge - the first step on the road to becoming a professional architect.
Many courses at Kingston University are vocational, which means that they are:
- are specifically designed to prepare you for your chosen career
- offer you the chance to gain valuable work experience in industry or commerce
- offer you the chance to gain valuable work experience in industry or commerce
4.Harvard University - graduate school of Design ( GSD )
Landscape architecture instruction at the GSD fosters students' inventiveness and creativity and cultivates the skills required for informed decision making. Landscape architects increasingly serve not only as designers, but also as advocates of landscape conservation. They are called upon to create landscapes that respond to the broad range of human habitation in diverse cultural and ecological contexts.
Students are encouraged to draw from historical precedents, design theory, civil engineering, and site analysis. Instruction also emphasizes the process of land planning, ecological analysis and the study of social, economic, legal, environmental, and policy issues affecting the whole design process.
The design studio provides the core of learning and inquiry; instruction and research emphasize critical analysis and a breadth of understanding in design, visual studies, theory, history, professional practice, and scientific research.
Snow Removal Services New Jersey


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December 23, 2010
THE PEOPLE, PLAINTIFF AND RESPONDENT,v.RYAN ROBERT WHITMAN, DEFENDANT AND APPELLANT.
Mendocino County Super. Ct. No. SCUK-CRCR-09-91776-002
The opinion of the court was delivered by: Kline, P.J.
P. v. Whitman CA1/2
California Rules of Court, rule 8.1115(a), prohibits courts and parties from citing or relying on opinions not certified for publication or ordered published, except as specified by rule 8.1115(b). This opinion has not been certified for publication or ordered published for purposes of rule 8.1115.
Ryan Whitman (appellant) was convicted, following a plea of no contest, of two counts of first-degree robbery of an inhabited dwelling in concert with at least two other persons. On appeal, he contends the trial court (1) abused its discretion when it imposed the aggravated term of imprisonment on the first count of robbery, and (2) violated the prohibition against the dual use of facts when it imposed a consecutive sentence on the second robbery count. We shall affirm.
Appellant was charged by felony complaint with two counts of first-degree robbery of an inhabited dwelling while acting in concert with at least two other persons (Pen. Code, §§ 211, 212.5, 213, subd. (a)(1)(A)--counts one and two);*fn1 one count of first degree burglary (§§ 459, 460, subd. (a)--count three); and four counts of false imprisonment (§ 236)--counts four through seven).
On September 18, 2009, appellant pleaded no contest to counts one and two: first-degree robbery of an inhabited dwelling while acting in concert with at least two other persons. The remaining charges were dismissed.
On December 18, 2009, the trial court sentenced appellant to the upper term of nine years on count one and a consecutive term of two years on count two.
On December 22, 2009, appellant filed a notice of appeal.
FACTUAL BACKGROUND*fn2
On January 7, 2008, at about 7:00 p.m., five or six men entered the house of appellant's former employer, Michael Stanley, in Mendocino County.*fn3 Michael told police that some of the men were armed and wearing ski masks. Some suspects went into other areas of the house, where Michael's wife and children were located. The men who remained with Michael restrained him with zip ties and assaulted him with a taser gun and the butt of a pistol. One of the men held his head down, and whenever he attempted to lift it, the man would hit him. In addition, the men kept asking him, " 'Where is it?' " and, " 'Where is the marijuana?' " While he was restrained, one of the suspects pulled down Michael's pants and tried to " 'stick a fork up my ass,' " and another threatened to shoot him in the knee if he did not cooperate. The men beat him whenever he did something they did not like.
The men then ransacked the house, taking items of value, including several televisions, cell phones, an X-Box 360, and other household electronic items. Before leaving, the men "hog-tied" Michael and his wife and put them in one of the children's rooms. The children were put in the bathroom with a chair in front of the door. The men then left in Michael's vehicles.
When asked why the suspects might have targeted his home, Michael initially denied that any marijuana or money was taken from him, and said that he was not in the marijuana business anymore. After he was told that a large sum of money was recovered from the suspects, Michael acknowledged he had had a large amount of money stashed in a tree stump about half a mile away from his house. He guessed the amount hidden there was between $30,000 and $40,000, from previous marijuana sales. He told police that he led the suspects to the stump and they took the money, before leaving.
Michael broke free of his restraints, went to his brother-in-law's house, and told him what had occurred. His brother-in-law and two other men said they had seen two vehicles--a blue-green sport utility vehicle (SUV) and a gold Nissan Sentra--parked in a clearing about a mile away from Michael's residence and thought they had looked suspicious. The four men then went searching for the suspects. They located the SUV just north of Willits and called the Sheriff's Office with the vehicle information.
Police observed that Michael had swelling on his head, a hematoma behind his right ear and another near his right eye. He also had marks on his lower back area and marks on his upper right buttocks, consistent with the teeth of a dining fork.
Michael's wife, Nichole, told police that she had been in her children's bedroom before going into the hallway and seeing a male wearing a black mask. The man pushed her back inside the bedroom. She eventually went into the kitchen, where she observed Michael tied up and being beaten by one of the men with a black object she thought was a taser. When she tried to look at the man's face, someone grabbed her by the hair and her face was forced towards the floor. She was then shown a taser and forced into a room with her two children, ages six and two. She was guarded by one of the men until, about an hour later, the men used duct tape to tape her to a chair next to her husband. Her children were locked inside the bathroom with a chair against the bathroom door. The men then fled. Nichole and Michael broke free and went to her brother's house.
After receiving a dispatch, police encountered the SUV on US Highway 101. A high-speed pursuit ensued until the SUV eventually ran over a spike strip that was placed in the roadway, causing three of the vehicle's tires to become deflated. The SUV then slowed to about 20 miles per hour and officers noted a rear door opening and closing several times, with someone throwing items from the vehicle. Eventually the SUV was stopped and the three occupants were arrested. Inside the SUV, police located multiple televisions, jewelry, ...
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- Create your own Stacking The Shelves post. You can use my official graphic or your own, but please link back to Tynga's Reviews so more people can join the fun!
- You can set your post any way you want, simple book list, covers, pictures, blog, sky is the limit!
- I am posting Stacking The Shelves on Saturdays, but feel free to post yours any day that fits you.
- Visit Tynga's Reviews on Saturday and add your link so others can visit you!
- Visit other participants link to find out what they added to their shelves!
Books Mentioned:
Paperbacks from Bookcloseout:
twenty boy summer
Jellicoe road
A Bad Boy Can Be Good for a Girl
Good Girls
Anatomy of a Boyfriend
Girl in the Arena
Hardcovers from Bookcloseout:
the beginning of after
So Shelly
The Body Finder
Desires of the Dead
Blood Magic
Dash and Lily's Book of Dares
The Julian Game
Bought at Barnes and Noble
Anna Dressed in Blood
Beauty Queens
My Life Next Door
Recieved From Book Blogger Friend Eileen
The Nine Lives Of Chole King


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Matthew Arnold
Matthew Arnold (1822-1888 / Middlesex / England)
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Epilogue To Lessing's Laocooen
One morn as through Hyde Park we walk'd,
My friend and I, by chance we talk'd
Of Lessing's famed Laocooen;
And after we awhile had gone...
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MILLBURN, NJ - This weekend, the Millburn Millers Varsity football team strapped on their pads for their season opener against the Livingston Lancers. The annual football game, which took place this weekend, resulted in a very different conclusion than last year's Millburn trouncing of Livingston. It was coach Patrick Leonardis second time seeing the Lancers. In his first year, Leonardis lead his Miller squad to a 32-0 defeat on Livingston's home turf. This year, though, in a very different outcome, the Lancers beat their Super Essex Conference enemy 21 to 14. This result is consistent with the Lancer trend of beating Millburn, which had been the case for the past four years, with the exception of last year.
Millburn got on the scoreboard early when junior quarterback, Jared Harris threw a touchdown pass to senior wide receiver, Gabe Greenberg in first quarter of the game to give the Millers a seven point edge. The Miller drive started when Millburn's Evan Steinfeld recovered a fumble to put the Millers in good field position for a score.
In the first half, there was an extended stoppage in play when Livingston's guard and defensive tackle, Ryan Distler went down with an undisclosed injury. In a show of true sportsmanship, both Millburn and Livingston players and coaches assisted Distler until medical help arrived. There is no further information on the injury thus far.
With 3:57 left in the second quarter, Millburn's second string quarterback Zack Weinstein was brought into the game after starting quarterback, Jared Harris' helmet came off in the preceding play forcing him to sit out a snap. On the snap, quarterback, Zack Weinstein fumbled the snap and the Lancers recovered. Livingston took advantage of this Miller misfortune just prior to the end of the first half. With 20.2 seconds left in the second quarter, Livingston's Mark Silvestre caught a pristine, twenty-five yard pass by quarterback Matt Gibbon, which tied up the game 7 to 7 going into halftime. Silvestre had a big day throwing 41 yards on two receptions. The Lancers tried to push their luck after their seven point gain by going for an onside kick, but Millburn recovered the onside kick with 16.9 seconds in the second quarter and was not caught off guard. The Millers were unable to score off of this opportunity, but the kick recovery gave them optimism going into the locker room at half time.
The Livingston Lancers took the lead for the first time in the game with 1:37 left in the third quarter when running back, Brandon Cohen ran it 20 yards into the end zone to give Livingston a 14 to 7 lead. Cohen ran the ball very well for the Lancers and had 102 yards on 12 attempts along with his third quarter score. On the following kickoff, Millburn's Gabe Greenberg returned the kick to the Millburn 47 yard line. Despite this fantastic return, once again Millburn was unable to convert on the ensuing drive.
Once the fourth quarter came around, Livingston made a quick effort to extend their lead. Number 24, Johnny Schriber ran in for the score to give Livingston 21 to 7 lead with seven minutes left in the fourth quarter.
The tide turned in Millburn's favor with 6:21 left in the fourth quarter when Jake Pickard, the Miller's punter bounced a punt off of a Livingston player, which allowed Millburn to jump on the ball and recover possession with great field positioning. On the succeeding play, Millburn's Evan Steinfeld caught a pass and ran it into the end zone to make the score 21 to 14, with 6:13 left in the game.
The Millers were trying to make a comeback and had a glimmer of hope until just over a minute left in the game when junior quarterback; Jared Harris threw an interception which helped to seal the Lancer victory.
The Livingston defense had a very impactful game. Not only did they hold the Millers to only two scores, but they achieved a total of 3 turnovers. Brandon Cohen had an interception along with 7 tackles. Johnny Schriber had 12 tackles, Zach Marini had a sack, and both Llewe Rocha and Ryan Distler recovered fumbles for Livingston.
Livingston quarterback Matt Gibbon had a fantastic performance Saturday afternoon. Gibbon found 7 receivers on 12 attempts and threw for 90 yards to help the Lancers edge out Millburn. Gibbon had both an interception and a touchdown during the game.
With the victory, the Lancers move to 1-1 on the year, while the Millers open up the season 0-1.The Millburn Millers look to obtain a better result next week when they face off against West Side at home on September 27th at 7:30 pm. The Lancers look to continue their hot streak at home next Saturday night versus Columbia High School. This victory was particularly welcome for Livingston after last week's 54 to 7 defeat to Lincoln High School in the Lancers home opener.