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Anti-inflammatory and anti-cancer activities of sterol rich fraction of cultured marine microalga Nannochloropsis oculata
Kalu Kapuge Asanka Sanjeewa, Ilekuttige Priyan Shanura Fernando, Kalpa W. Samarakoon, Hetti Handi Chaminda Lakmal, Eun-A Kim, O-Nam Kwon, Matharage Gayani Dilshara, Joon-Baek Lee, You-Jin Jeon
DOI : https://doi.org/10.4490/algae.2016.31.6.29
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Sanjeewa, Fernando, Samarakoon, Lakmal, Kim, Kwon, Dilshara, Lee, and Jeon: Anti-inflammatory and anti-cancer activities of sterol rich fraction of cultured marine microalga Nannochloropsis oculata
Algae 2016; 31(3): 277-287.
Published online: September 30, 2016
DOI: https://doi.org/10.4490/algae.2016.31.6.29
Kalu Kapuge Asanka Sanjeewa1, Ilekuttige Priyan Shanura Fernando1, Kalpa W. Samarakoon2, Hetti Handi Chaminda Lakmal1, Eun-A Kim1, O-Nam Kwon3, Matharage Gayani Dilshara4, Joon-Baek Lee5, You-Jin Jeon1,*
1Department of Marine Life Science, School of Marine Biomedical Sciences, Jeju National University, Jeju 63243, Korea
2Industrial Technology Institute, Colombo 7, 00700, Sri Lanka
3Marine Biology Center for Research and Education, Gangneung-Wonju National University, Gangneung 25457, Korea
5Department of Earth and Marine Sciences, College of Ocean Sciences, Jeju National University, Jeju 63243, Korea
*Corresponding Author: E-mail: youjinj@jejunu.ac.kr, Tel: +82-64-754-3475, Fax: +82-64-756-3493
Received April 18, 2016 Accepted June 29, 2016
Copyright © 2016 The Korean Society of Phycology
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Five fractions separated from Nannochloropsis oculata using solvent-solvent partition chromatography of 80% methanolic extract of N. oculata (NOM) followed by the open silica column chromatography of its hexane fraction (NOMH) for the anti-inflammatory on RAW 264.7 cells and anti-cancer activities on HL-60, A-549, HEP-3B, HCT-116, and SW-480 cancer cells. All the five fractions showed potential anti-inflammatory activities against lipopolysaccharide-stimulated RAW 264.7 macrophages cells with IC50 values less than 6.25 μg mL−1. Moreover, 90% n-hexane column elution of NOMH (NOMH90) down-regulated lipopolysaccharide-stimulated protein levels of inducible nitric oxide synthase and cyclooxygenase-2. Furthermore, NOMH90 showed marked cytotoxic effect on the HL-60 cells with IC50 value of 23.58 ± 0.09 μg mL−1. In addition, Hoechst 33342 cell permeable dye used to visualize the apoptosis nucleus and cell cycle analysis measured Sub-G1 DNA contents to confirm reduction of the cell viability in NOMH90 treated cells due to induction of apoptosis in HL60. These results are quite related to the phytosterol contents of the NOMH fractions and the results suggest N. oculata extracts might be useful as potential sources of natural anti-inflammatory and anti-cancer compounds. In conclusion, the sterol content in N. oculata might provide a promising role in future medicines in anti-inflammatory and anti-cancer.
Key words: anti-cancer; anti-inflammation; hexane fraction; Nannochloropsis oculata; sterol
Sterols are one of the important component in the membrane of all eukaryotic cells (Volkman et al. 1998). Moreover, cholesterols are major sterols of vertebrate organisms and mixture of various phytosterols containing in photosynthetic organisms (Lopes et al. 2013). Farnesyl diphosphate convert into squalene and this step activate the isoprenoid pathway which capable of producing sterols in biological systems (Hartmann 1998). Sterols located in microalgal cell membranes act as biomarkers for identifying organic matters in their surroundings and acting as an essential component for cell proliferation (stigmasterol) and signal transduction in microalgae (Hartmann 1998, Volkman et al. 1998). Moreover, phytosterols separated from marine microalgae found to possess interesting bioactive properties, including anti-inflammatory, anti-cancer, antioxidant, antidiabetic, and neuromodulatory activity (Yasukawa et al. 1996, Francavilla et al. 2012, Kim et al. 2014, Luo et al. 2015).
Cancer developed in multicellular organs when the balance between the cell generation and cell death is altered (Cotter 2009). Cancer is considered as one of the major threats to human thus been made a high level of suffering and economic losses to entire humans. Due to the high level of impact, considerable attention has been made to eliminate cancer (Jayaprakasam et al. 2003). Some reports suggest that there has been a positive relationship between inflammation and tumor progression with anticancer compounds (Coussens and Werb 2002). Most of the anticancer drugs developed to treat cancer have been shown to induce apoptosis in cancer cells. Moreover, those anticancer drugs induce apoptosis cell death via, inducing the endonucleolytic cleavage of DNA and changes in chromatin condensation of cancer cells (Hickman 1992). Furthermore, phytosterols separated from microalgae have identified as an excellent source for develop chemopreventive drugs for cancer diseases. Moreover, a number of studies have reported phytosterol separated from microalgae induce apoptosis cell death in cancer cells (Luo et al. 2015).
Inflammation can define as the normal protective response of living tissue to injury or infection. However, an inflammatory response is a series of local cellular and vascular responses that immediately occurs following tissue damage as a result of either, physiological or pathological processes and act as a protective mechanism of organisms to remove the noxious stimuli from the tissues and to initiate the healing process (Calder 2006, Medzhitov 2008, Huang et al. 2012).
During inflammation, inflammated cells start to produce pro-inflammatory cytokines as well as stimulate the expression of nitric oxide synthase isoform and an inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Furthermore, expression of iNOS led to produce tiny, reactive radical gas known as nitric oxide (NO) in phagocytes including macrophages and dendritic cells and COX-2 led to produce protaglandin E2 (PGE2) in lipopolysaccharide (LPS)-stimulated RAW cells (Nathan 1992, Lowenstein et al. 1993, Gordon 2002, Lee et al. 2002).
Nannochloropsis species belong to the family eustigmatophyte and are photosynthetic eukaryotic unicellular microorganism's habitats in both fresh and sea water environments. Moreover, eustigmatophyte microalgae species are popular as a good source of phytosterols (Sukenik et al. 1993, Converti et al. 2009). However, knowledge on bioactive properties of Nannochloropsis species are limited and very low. The aim of this study was to investigate the effects of partially purified sterol-rich hexane fraction of marine microalga Nannochloropsis oculata on anti-cancer and anti-inflammatory activities.
Chemicals and reagents
Five cancer cell lines including human promyelocytic leukemia cell line (HL-60), human colon adenocarcinoma cell line (SW-480), adenocarcinomic human alveolar basal epithelial cell line (A549), hepatocellular carcinoma cell line (Hep-3B), and colorectal carcinoma cell line (HCT-116) were used to evaluate anti-cancer activity and murine macrophage cell line (RAW 264.7) was used to measure in vitro NO production and African green monkey kidney (Vero) cell line was used to evaluate the cytotoxic effect of tested concentrations. All cell lines were purchased from the Korean Cell Line Bank (Seoul, Korea). Roswell Park Memorial Institute (RPMI-1640) medium, Dulbecco's modified Eagle's medium (DMEM), penicillin-streptomycin and fetal bovine serum were purchased from Gibco/BRL (Burlington, ON, Canada). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich (St. Louis, MO, USA). All other chemicals and reagents used in these experiments were of analytical grade.
Culture of marine microalga
Marine microalga N. oculata was provided by the Korea Marine Microalgae Culture Center. The alga was pre-cultivated in 5 L glass vessels (medium 4 L) and then inoculated in 30 L plastic cylinders at 20°C, although air was supplied continuously at 5 L min−1 by air-lift. Sixty watt fluorescent lamps were used to provide light energy at an intensity of 34 μmol photons m−2 s−1 throughout the growing period. The microalga was cultured in Conway medium as previously described by Walne (1966). In brief, medium was prepared from filter-sterilized seawater and the culture was continuously active during the 8 to 10 days after inoculation. The cells were flocculated with 200 ppm Al2(SO4)3 (v/v) (Magicpool-99; Ilshin Biochemical, Hanam, Korea), and then recovered with centrifugation at 2,000 rpm using a basket centrifuge (Hanseong Co., Ansan, Korea). The harvested N. oculata biomass was frozen at −80°C and preserved until freeze dry.
Determination of proximate chemical compositions of Nannochloropsis oculata
Proximate composition of moisture, protein and ash contents were determined using standard Association of Official Analytical Chemists (AOAC, 1998) methods (Cunniff 1998). Crude protein was measured by calculating a conversion factor of 6.25. Crude lipid content was measured by using Soxhlet method with diethyl ether solvent (Soxtec 2050; FOSS Analytical, Hillerød, Denmark) and crude ash was determined by incineration of the sample at 600°C in the muffle furnace (B180; Nabertherm GmbH, Lilienthal, Germany). Moisture content was determined by the oven-drying method at 105°C in the moisture analyzer (mb45; OHAUS, Nänikon, Switzerland).
Sample preparation and fractionation
The freeze dried N. oculata sample (10 g) was extracted three times at less than 40°C for 90 min with 80% methanol solution (1.0 L) using an ultrasonic bath (JAC 2010; Kodo Technical Research Co. Ltd., Hwaseong, Korea). The liquid layer was obtained via vacuum filtration, and the filtrates were concentrated using rotary evaporator followed by freeze dryer. The freeze dried powder was considered as the crude methanolic extract of N. oculata (NOM).
Then the NOM powder was dissolved in distilled water and partitioned according to the polarity using n-hexane (NOMH), chloroform (NOMC), and ethyl acetate (NOME), respectively. The resulting solvent fractions were concentrated using rotary evaporator and freeze-dried. Further fractionation of an active fraction (NOMH) was conducted using an open silica column by hexane / ethyl acetate step-gradient elution with successive ratios of 90 : 10 (NOMH90), 80 : 20 (NOMH80), 70 : 30 (NOMH70), 60 : 40 (NOMH60), and 50 : 50 (NOMH50). Each fraction was evaporated by using a rotary evaporator (Fig. 1).
Total sterol content measurement of Nannochloropsis oculata extracts
Total sterols contents of N. oculata extract and its fractions ware assessed by cholesterol quantification using Liebermann-Buchard reaction with some slight modifications (Huang et al. 1961, Costa et al. 2013). Briefly, 150 μL of Liebermann-Burchard reagent (Burke et al. 1974) were mixed with 5 μL of each sample (in a 96-well plate) followed by incubation at 37°C for 30 min. Absorbance was measured at 625 nm in a spectrophotometer and the sterol concentration was determined from a standard curve for cholesterol content.
Cell experiments
Vero cells and all cancer cell lines were grown in RPMI medium, while RAW 264.7 cells were cultured in a DMEM medium. Both culture mediums were supplemented with 10% heat-inactivated FBS and 1% streptomycin (100 μg mL−1), and penicillin (100 unit mL−1). The cells were maintained and incubated in modified atmospheric condition which contained 5% CO2 and at 37°C (ambient temperature). Cells were sub-cultured within 2 days intervals and exponential phase cells were used for experiments.
Determination of cytotoxicity on Vero cells and anti-cancer activities of extracts
MTT assay, is a test of metabolic competence predicated on the assessment of mitochondrial performance, which depends on the reduction of the yellow color tetrazolium salt MTT into its blue-black formazan crystals via mitochondrial succinate dehydrogenase in viable cells (Pieters et al. 1988). Cell cytotoxicity of all cell lines were estimated via a colorimetric MTT assay as described by Mosmann (1983), with slight modifications. Briefly, 190 μL of cells were seeded in a 96 well plates with 1 × 105 cells mL−1 concentration and plates were incubated for 24 h, and then treated with 10 μL from measured concentration (6.25, 12.5, or 25 μg mL−1) of NOMH fractions and again incubated for 24 h. Ten microliter of MTT reagent (2 mg mL−1 in phosphate buffered saline [PBS]) was added to each well and aspirate the medium after 3 h of incubation and formazan crystals were dissolved in DMSO, and then the amount of blue-black formazan was measured using enzyme-linked immunosorbent assay (ELISA) reader at 540 nm.
Determination of NO production
NO production was assayed using a method described by Leiro et al. (2002) with some modifications. On the basis of nitrate production in RAW 264.7 culture supernatants, measured with Griess solution. Briefly, 450 μL of cell suspensions (1 × 105 cells mL−1) were incubated on 24 well plates for 24 h and then treated with 25 μL of measured concentrations of NOMH fractions to each well and further incubated for 1 h and treated with 25 μL of LPS and further incubated for 24 h. Then 100 μL cell suspensions were mixed with equal volume of Griess reagent. After 10-min incubation, the absorbance was read with an ELISA reader at 540 nm.
Western blot analysis
Various concentrations of (3.12, 6.25, and 12.5 μg mL−1) the active fraction were used to determine anti-inflammatory protein expression on RAW 264.7 cells. Proteins were extracted with PRO-PREP protein extraction kit (iNtRON Biotechnology, Seongnam, Korea). Briefly cell lysates were centrifuged 16,000 rpm at 4°C for 20 min to collect supernatants. The protein concentration of supernatants were determined by using Bio-Rad protein assay kit (Bio-Rad, Hercules, CA, USA). The cytoplasmic proteins (40 μg) were electro-transferred onto a nitrocellulose membrane following separation on a 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis under denaturing conditions. After blocking with 5% nonfat milk for 1 h, the blots were separately incubated with specific primary rabbit polyclonal anti-rabbit iNOS or mouse monoclonal anti-mouse COX-2 or β-actin (Cell Signaling Technology, Beverly, MA, USA) antibody for 60 min and washed twice with Tween 20/Tris-buffered saline (TTBS). Then, the blots were incubated with horseradish peroxidase–conjugated anti-mouse or anti-rabbit IgG for 45 min followed by visualized using an enhanced chemiluminescence (ECL) reagents (ECL; Amersham, Arlington Heights, IL, USA) onto a nitrocellulose membrane.
Nuclear staining with Hoechst 33342
HL-60 cells were cultured in 24-well plates at a concentration of 1 × 105 cells mL−1 and cells were incubated at 37°C for 24 h. The cells were then treated with various concentrations of NOMH90 (6.25, 12.5, and 25 μg mL−1) and incubated for an additional 24 h again at 37°C. Then, Hoechst 33342, a DNA-specific fluorescent dye was added to the culture media at a final concentration of 10 μg mL−1 and the plates were incubated for an additional 10 min at 37°C. The stained cells were then observed under a fluorescence microscope equipped with a CoolSNAP-Pro color digital camera in order to determine the degree of nuclear condensation.
Cell cycle analysis
HL-60 cells at a density of 1 × 105 cells mL−1 were treated with various concentration of NOMH90 (6.25, 12.5 and 25 μg mL−1) or the solvent alone. After 24 hours, the cells were harvested by centrifugation, washed twice with PBS and suspended in 1 mL 70% ethanol solution at 4°C for 30 min. After 30-min incubation cells were centrifuged, and the cell pellets were stained with 4′,6-diamidino-2-phenolindole (DAPI). DNA content was then analyzed with a FACSCalibur flow cytometer (Becton Dickinson, San Jose, CA, USA).
All the data were expressed as mean ± standard deviation of three determinations. Analysis of variance was carried out for the collected data using the Statistical Package for Social Science (IBM SPSS Statistics for Windows, version 20.0; IBM Corp., Armonk, NY, USA) statistical analytical package. Mean values of each experiment were compared using one-way analysis of variance. The Duncan's multiple range test was used for the mean separation. A p-values less than 0.05 was considered significant.
Proximate chemical composition and sterol contents of the microalgal samples
The proximate composition and total sterol content of N. oculata were investigated. Total protein content was recorded as 30.52% dry weight basis (DW). Ash and lipid contents were respectively recorded as 30.64% and 7.99% DW (Table 1). Sterol contents of the algal extract and its fractions were measured using Liebermann-Buchard reaction (Table 2). According to the results, sterol content of NOM was recorded as 19.38% and phytosterols contents of its solvent fractions (NOMH, NOMC, and NOME) ranged between 32.35 to 11.36% and the highest was recorded from NOMH fraction. As well as bioactivities of NOM fractions were also increased with sterols contents. Hence, study was continued only with NOMH fraction. Furthermore, five fractions eluted from NOMH fraction by using open silica column were also measured for the sterol content. The sterol contents of the five NOMH fractions were ranged between 6.34 to 63.59% and the highest was recorded from the NOMH90 while the lowest from NOMH50 fraction.
Cytotoxicity assay
The cytotoxicity study was carried out for each NOMH fraction to measure optimum sample concentration for the growth of Vero cells. The effects of NOMH fractions on the cell viability were determined with the MTT assay (Fig. 2). According to the cell viability results, all the tested fractions did not exhibit cytotoxic effect on the Vero cells, except 25 μg mL−1 concentration of NOMH80 (92% cell viability), which showed a significant cytotoxicity on the cells compared to the control (Fig. 2). Therefore, it revealed that all the fractions tested concentrations are not cytotoxic to the cells.
Evaluation of anti-inflammatory activity of NOMH fractions on RAW 264.7 macrophages cells
To investigate the anti-inflammatory effects of the five NOMH fractions, the inhibitory effects on NO production were examined in LPS activated RAW 264.7 macrophage cells. All the five fractions inhibited NO production on macrophages cells in a dose-dependent manner, compared to the LPS treated cells as shown in Fig. 3A.
RAW 264.7 cell viability in the presence of NOMH fractions are shown in Fig. 3B. According to the results NOMH50 and NOMH90 fractions were not significantly affected on cell viability of RAW 264.7 cell. However, NOMH60, NOMH70, and NOMH80 fractions at the 25 μg mL−1 exhibited significant cytotoxicity. According to the results, NOMH90 fraction has been subjected to Western blot analysis in which NOMH90 fraction down-regulated the iNOS and COX-2 protein expression in a dose-dependent manner as shown in Fig. 3.
Anti-proliferative effect of NOMH fractions against the growth of selected five cancer cell lines
Cell proliferation is a crucial marker for the promotion and progression of carcinogenesis. In this study we measured cell viability percentage of five respective cancer cell lines HL-60, A-549, HEP-3B, HCT-116, and SW-480 incubated with different concentrations of NOMH (Fig. 4). According to the results NOMH80 and NOMH90 showed strong anticancer effect against HL-60 cells in dose-dependent manner. Moreover, according to the results IC50 values of NOMH80 and NOMH90 on HL-60 cells were 24.81 ± 0.34 and 23.58 ± 0.09 μg mL−1, respectively.
Apoptosis morphological changes observed under fluorescent microscopy
Apoptosis can be defined as breakdown of cell DNA into the small size basal points (180–200) or 50–300 kb ranged large DNA fragments (Lizard et al. 1995), hence cells with homogeneously stained nuclei were considered viable, whereas the presence of chromatin condensation or fragmentation was indicative of apoptosis. Fig. 5 noticed that NOMH90 dose-dependently increased the proportion of apoptosis body formations in HL-60 cells.
Induction effect of Sub-G1 contents in HL-60 cells by NOMH90
Anti-cancer effect of NOMH90 evaluated by measuring Sub-G1 DNA content population percentage of the HL-60 cells after incubated with different concentrations of NOMH90 for 24-h. According to the results, NOMH90 fraction showed cell cycle arrest and accumulation of cells in the Sub-G1 phase in a dose-dependent manner (Fig. 6). Moreover, the apoptosis body formations corresponded with the accumulation of Sub-G1 content and further were determined as 33.2, 56.7, and 61.2% of Sub-G1 population against the NOMH90 concentrations (6.25, 12.5, and 25 μg mL−1), compared to the control, respectively (Fig. 6). Thus, we can suggest that NOMH90 has potential to induce cells death in HL-60 via inducing apoptosis in the nucleus.
Currently many studies focus on bio-diesel production and / or extract from the marine microalga N. oculata, (Converti et al. 2009, Umdu et al. 2009, Van Vooren et al. 2012) but limited interest on bioactivities of extracts from N. oculata microalga. The present study was undertaken to examine the anti-inflammatory effect of N. oculata extract and its sterol-rich fractions on LPS-stimulated RAW 264-7 cells and the anticancer effect on five cancer cell lines. We found that NOMH90 dose-dependently reduced the NO production in the cells and showed anticancer effect on HL-60 cancer cell line (Fig. 4). Therefore, we measured apoptosis body formation and cell cycle analysis only for NOMH90 using HL-60 cells. According to the apoptosis results and cell cycle analysis it clearly demonstrated that the reduction of cell viability of HL-60 cells are due to the cell death via induction of apoptosis pathways. Instead of above observations we found that anti-inflammatory and anti-cancer activities are quite similar to the sterol contents of the NOMH fractions.
Macrophage cells are considered as the first line of host defense against inflammation. When macrophage cells expose to the stimulating agents such as LPS, they start to express inflammatory responses to act against inflammatory stimuli (Smith 1994, Kim et al. 2006, Ma and Quirion 2008). iNOS and COX-2 are known as respective inflammatory mediators involved in the pathogenesis of diseases associated with inflammation (Lee et al. 2003, Kim et al. 2009). Expression of iNOS proteins induce the NO production and COX-2 induce the PGE2 production in LPS-activated macrophages cells (Han et al. 2004, Greenhough et al. 2009). Moreover, NO is a labile gaseous free radical as well as NO can act as a neurotransmitter, vasodilator and in cellular level defense against tumor cells, parasites, and bacteria in the immunological system (Nakagawa and Yokozawa 2002). Besides excess and prolong production of NO cause pathogenesis of inflammation, and cancer (Kobuchi et al. 1997). PGE2 is also capable to regulate platelet aggregation, vascular permeability, and thrombus formation in the development of inflammatory responses (Knöferl et al. 2001). Therefore, it is important to control the expression of iNOS and COX-2 to control inflammatory responses. Recently, number of studies confirmed that the compounds and solvent fractions separated from microalgae have potential to use as anti-inflammatory agents (Kim et al. 2013, Somchit et al. 2014). Instead of that, Yasukawa et al. (1996) reported that the sterols isolated from Chlorella vulgaris have promising activity against 2-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice (Yasukawa et al. 1996). Our present results also, indicated that NOMH90 (sterol-rich fraction) inhibited the NO production in LPS-stimulated RAW 264.7 cells. The anti-inflammatory effects of NOMH90 were accomplished by the decrease in expression of iNOS and COX-2 proteins in cytosol dose-dependent manner. Thus, there will be a potential to develop anti-inflammatory drugs using N. oculata microalga.
Keeping a balance between cell proliferation and cell death is an important mechanism to maintain homeostasis in multicellular organisms (Vanden Berghe et al. 2013). Induction of apoptosis is one major form of cell death and it can be observed from embryonic development through immunological regulation. Also, apoptosis is a useful tool to eliminate superfluous, inappropriate or harmful cells from the infected site in case of pathological situation (Jacobson et al. 1997). Deregulated levels of apoptosis in many cancer cells is a common problem to eliminate cancer cells in an effective manner. Therefore, the compounds that are capable of restoring apoptosis is known to be a promising strategy for cancer prevention (Fesik 2005). Justo et al. (2001) reported anticancer effect of extract collected from microalga Chlorella vulgaris. Further, their results demonstrated prolonged the survival of mice inoculated with the Ehrlich ascites tumor after administration of C. vulgaris extracts on Ehrlich ascites tumor infected mice compared to the control (Justo et al. 2001). According to our results (Figs 5 & 6) it is also indicated that NOMH90 fraction has great impact on inducing apoptosis in HL-60 cells in a dose dependent manner.
In conclusion, the present study highlighted the anti-inflammatory and anticancer activity of partially purified sterol-rich fraction isolated from 80% methanol extract and followed by open silica column chromatography of N. oculata microalga. According to the results of NOMH90 fractions of N. oculata microalga may be a feasible therapeutic or preventive candidate for developing drugs to treat diseases associated with inflammation and Human promyelocytic leukemia cancer.
This study was supported by the 2016 scientific promotion program funded by Jeju National University.
Extraction and fractionation scheme of the Nannochloropsis oculata fractions.
Cytotoxic effect of NOMH fractions against the growth of Vero cells. Cells were incubated with different concentrations (6.25, 12.5, and 25 μg mL−1) of NOMH fractions for 24 h and viability of the cells were measured using colorimetric 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Data points and bar represent arithmetic means ± standard deviation (n = 3). Means with same letters are not significantly different at 0.05 sigma level.
Anti-inflammatory effects of the NOMH fractions. (A) The dose-dependent inhibition of the NOMH against nitric oxide (NO) productionin lipopolysaccharide (LPS)-challenged RAW 264.7 macrophages. (B) The cytotoxicity of those NOMH fractions on RAW 264.7 cells in the presence of LPS. (C) The inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) inhibition of NOMH90 in dose dependent manner. Cell lysates (40 μg) were resolved on sodium dodecyl sulfate–polyacrylamide gels and then transferred to nitrocellulose membranes, and probed using antibodies against iNOS and COX-2. Data points and bar represent arithmetic means ± standard deviation (n = 3). The intensity of the bands was measured by ImageJ software. Relative amounts of iNOS and COX-2 compared to β-actin. The gel shown is a representative of the results from three separate experiments. Means with same letters are not significantly different (p > 0.05).
Anti-proliferative effect of the NOMH fractions against the growth of A-549 (A), HEP-3B (B), HCT-116 (C), SW-480 (D), and HL-60 (E) cancer cells. Cells were incubated with different concentrations (6.25, 12.5, and 25 μg mL−1) of NOMH fractions for 24 h and viability of the cells were measured using colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Data points and bar represent arithmetic means ± standard deviation (n = 3). Means with same letters are not significantly different at 0.05 sigma level.
Effect of NOMH90 on morphological changes in HL-60 cancer cell line. Cells were treated in the absence of NOMH90 (A) or in the presence of 6.25 (B), 12.5 (C), and 25 μg mL−1 (D) of NOMH 90 for 24 h, stained with Hoechst 33342, and observed by fluorescence microscopy. Arrows (B–D) indicate a typical apoptosis body.
Flow cytometric analysis of DNA cleavage in NOMH90 treated cells. Cells were incubated with various concentrations of NOMH90 for 24 h and stained with 4′,6-diamidino-2-phenolindole (DAPI) for DNA content analysis. Percentages of apoptosis cells are represented in each panel. Data in the figure represent percentages of Sub-G1 cell populations in cell cycle distribution. CON, control.
Proximate chemical compositions of the marine microalga Nannochloropsis oculata on dry weight basis
19.56 ± 0.37 30.52 ± 0.04 7.99 ± 0.29 30.64 ± 0.22 11.29 ± 0.26
Values are mean ± standard deviation (n = 3).
Analysis was according to the Association of Official Analytical Chemists (AOAC, 1998) methods.
Sterol contents of Nannochloropsis oculata fraction
Fraction name
Sterol content (%)
NOM 19.38 ± 0.87
NOMH 32.35 ± 1.75
NOMC 29.26 ± 0.87
NOME 11.36 ± 3.49
NOMH50 6.34 ± 0.58
NOMH60 38.02 ± 1.16
Values are presented as mean ± standard deviation (n = 3).
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Results 21 - 30 of 621 for Ocean Energy
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redpajama_set_name: RedPajamaCommonCrawl


Is there any way to generate PDF files from classic ASP? I have a bunch of user-entered data that needs to be turned into a PDF that the user can download. how to make preview pane show pdf files clicks the Generate button and gets branded PDF invoice created. Image below demonstrates Image below demonstrates the running app main window with entered information.
To create a new ASP.NET Web Application-project and a default Web Form you have to follow the following steps: 1. In Visual Studio .NET, on the Start Page, click New Project.
You must have a Common Gateway Interface (CGI) application on the web server to collect and route the data to a database. Any existing CGI application that collects data from forms (in HTML, FDF, or XML format) can be used.


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Review: "City On The Edge Of Forever" Remastered [w/ new pics & vids]
| October 12, 2006 | By: Dennis Russell Bailey 39 comments so far
Captain Kirk Meets The No-Win Scenario
The Enterprise crew discovers an alien time machine on an uncharted, lifeless world. In a fit of drug-induced paranoia (it's an accident, kids — I can't stress this enough; don't do drugs, stay in school), "Bones" McCoy leaps back into 1930s New York and alters history such that the "Star Trek universe" never happens. Kirk and Spock go back in time themselves in order to fix history; in a twist familiar to anyone who has read much of Harlan Ellison's work, a blameless young woman who loves the protagonist has to be killed in order to make Everything Okay Again.
Shatner shows Kirk's depth
Flippant synopsis aside, I'm one of those geezers who consider this episode to be the finest "Star Trek" story ever produced. If asked by someone who knew nothing abut "Star Trek" to recommend an episode to demonstrate what all the fuss is about, this is the one I'd choose to show them. It's bereft of Klingons, Romulans, fake navy combat in space, straining warp engines and Spock/McCoy bickering. Kirk doesn't solve the problem by dint of superior wit or ethics or firepower. No Federation /America Uber Alles here. The choice Kirk has to make and the resulting denouement are tragic in the purest sense of the word; it's a moment in which only character and values matter and no good outcome is possible.
The power of the drama turns almost as much on performance as on the story, here. DeForest Kelley, whose character is in many ways the most vulnerably human of the Trek characters, is fragile in the early going and at his most laconic and charming during McCoy's "recovery." Nimoy displays the restrained empathy, at one time both studied and seemingly unconscious, that is emblematic of Spock. Most crucial, though, are two performances that can be summed up in a couple of phrases that look kind of wrong in cold print: Joan Collins' modesty and innocent appeal as Keeler and William Shatner's understated performance as Kirk. One is reminded again — as in watching the recent broadcasts of "Balance Of Terror" and "Devil In The Dark" — that Kirk's "iconic" status is more grounded in the thoughtful work Shatner did in the first year of "Star Trek" than in the often-parodied, exaggerated mannerisms and telegraphed delivery he would habitually slide into later in the series' run.
The Remastering is superb
As with the previously broadcast Remastered episodes, the live-action sequences are beautifully clear and newly color-balanced. The overall image runs a little less blue than the standard-resolution DVD releases; Kirk's accurately colored avocado gold uniform takes some getting used to after forty years of seeing it as yellow-gold.
The trimming of the episodes for running time is thoughtfully done. That can't be an easy thing to do, considering that six to seven minutes must be excised from each one in order to fit the commercial demands of current day broadcast.
A mixed bag on the CGI changes
Let's get the worst out of the way up front: the nacelle lighting effects on the CG Enterprise model remain terrible. Most everyone seems to agree on this except for the folks at CBS Digital. The effect used doesn't resemble anything in the original series however similar the mechanics of creating it may be. I've seen a couple of professional recreations and several fan efforts in CG that are more successful.
The poor effect is least annoying when the Enterprise model is seen in bright light and most garish when the ship appears in shadow. That's a shame, because the orbital shots of the Enterprise in the remastered "City" are largely in shadow. The scene lighting itself is a marvelous touch — it suits the mood of the episode, and it demonstrates an advantage of the technology being used here: the artists can use the familiar "stock shots" of the ship from episode to episode yet give them distinctive looks based on different environments each week.
CBS still can't get those nacelles right (click to enlarge)
The opening shot of this episode would have been a great example of how effective this could be — except that it's just about ruined by the far too-bright, frenetically spinning nacelles. I can't fathom how the people in charge look at this effect week after week and say "that's good enough."
The original globe of the Guardian's Planet is replaced, of course. So far, the CG folks have been doing a bang-up job of capturing the slightly impressionistic quality of TOS planets while enhancing the specific details of terrain, cloud and atmosphere. The Guardian's planet is no exception. The colors seem colder overall and are less monochromatic than the original.
Originally, Act One opened with an often-used pair of shots — one of the Enterprise approaching camera around the limb of a planetary globe, cutting to a reverse angle of the ship sailing away from camera. The Remastered version neatly combines these into a single shot. The "camera" tracks the Enterprise and follows it through the arc of its orbit. It's a good shot — the nacelles aren't too distracting (here, they even show some of the internal lighting and color variations that they should).
Something similar was done as the final shot of "Devil in The Dark," but it's still new enough to be unexpected, and it's delightful. That said, given the reverential approach that the artists are taking to this project I imagine they'll reach the bottom of this bag of tricks pretty quickly. Will there be any novelty left to the project, by the time they've done thirteen episodes or so, without a more radical rethinking of these kinds of shots? I guess we'll find out.
Lighting aside, the CBS Digital model of the Enterprise looks great. On screen it exhibits about the same amount of detail as the original 11-foot practical model, though publicity images that have been released show that there's somewhat more detail built into the textures than that. I'm not viewing these episodes in HD; perhaps more surface detail is visible at that higher resolution.
Very little else is changed in Act One. There's another nicely lit orbital flyby, and the final shot panning up from Kirk and into empty space has been tweaked. A deep blue atmospheric gradient has been added so that the transition from the foil-and-plaster "rock" background into the blackness of space is less abrupt.
Act Two opens with a new CG shot of the Guardian's planet. The original featured a camera push-in with the globe growing rapidly in frame for several seconds. The push-in on the new version is more subtle, but whereas before the planet itself rotated v-e-r-y slowly toward the west, it now is spinning at what appears to be about three hundred miles per hour and in a conventional Earthlike easterly direction. Perhaps it's a very small world (say about the size of Connecticut), but the speed of the rotation is jarringly unrealistic.
Hold on tIght when on the Guardian Planet (Click for WMV Video)
There are no other new enhancements until the scene in which Spock discovers the recorded newspaper headlines concerning Edith Keeler's two possible destinies. Here, new interference patterns have been created for the tricorder screen. The first is a denser and subtler version of the vertical black-and-white "static" originally used. The second, when the circuits burn out, is now a full-color effect that doesn't resemble video static.
They've done something else here that shows a fine interest in the details: where the "newspaper clippings" were originally badly matted onto the non-functioning "screen" of the tricorder, the inserted image has been enlarged slightly and properly matted so that there are no irregular black gaps around it. The brushed-aluminum face of the tricorder itself is even made to cast a credible shadow onto the edges of the "video screen."
Spock's Original Tricorder & TC2.0 (click to enlarge)
That improved effect is also used in Act Three, as Spock and Kirk review the divergent history caused by Keeler's survival, and is the only new effects shot in this act. The scene in which the bum who finds McCoy steals the Doctor's phaser and accidentally immolates himself was cut from the broadcast in order to shorten the episode. It's one of the most memorable effects moments in TOS, but I guess we'll have to wait for the Remastered HD-DVDs to find out if anything's been done to it.
There's one final improvement, in the last shot of the episode: the fog rolling off of the Guardian continues to do so during the final crew beam-out and during the end-of-story credits. Originally, the shot went to freeze-frame during the beaming and every time a credit was displayed.
Guardian: Smoke em if you got em (click for WMV Video)
Some things are best left alone
This kind of thing should be noted, I think, because it speaks as much to the taste and intentions of the people supervising the Remastered episodes as do the new effects themselves. This week, they resisted what had to be quite a temptation to extend the limited Guardian stage set with digital mattes.
I applaud that. There's something essential to appreciating what "Star Trek" really was and is, in that moment when William Shatner plants his feet on a sandy stage surrounded by a few phony broken Greek pillars, looks past the camera as if into the dark distance and matter-of-factly states: "These ruins extend to the horizon."
The original "Star Trek" asked a lot from the collective imagination of its audience, engendering a sense of participation that probably accounts for much of its enduring fascination. If you can't see those ruins in your mind when Kirk says that, this may not be the kind of trip you'll enjoy.
I've had my own "vision" of those ruins for forty years and am a little relieved not to have that emotionally thrilling and mysterious landscape overlaid with the very specific visualizations of a modern artist, however talented he or she might be.
The timeless look of the Guardian Planet (click to enlarge)
No credit where credit is due?
The show's closing titles are not changed at all. This bothers me. The artists who are creating new effects for these shows are certainly entitled to credit. They are working on very short deadlines, and in my opinion they're succeeding in the challenge in almost every instance. Certainly this lack of credit isn't an accidental oversight. Whatever the reason is for it, I hope that it will be addressed and corrected sooner rather than later.
Click for More Screenshots and vid
Conventions/Events/Attractions
Paramount Theme Park In China Featuring Star Trek-Themed Area Moving Forward
List, Trek Franchise
Celebrate Halloween With Characters From Each Star Trek Series In Dress-Up
Books, TOS
Book Review: 'Star Trek – A Celebration' Delivers On Its Promise And Then Some
Star Trek Universe TV
'Star Trek: The Pod Directive' New Official Podcast Launching This September
Semaj
I'm glad I'm not the only one to notice the fog from the Guardian not freezing over the credits like in the original. It was a nice touch.
Excellent review Dennis!
A good review for the quintessential Star Trek episode.
I think there's so much we take away from Star Trek on a personal level, some of us love the space navy stuff, the McCoy/Spock bickering, etc. But here, Dennis is right to say this story is about a moral choice, not about "winning" or triumphing over an enemy. Of course, it is not clear how long Kirk and Keeler knew one another before they final resolution took place, but I've come to consider that they spent about 3-4 weeks in one another's company. That may not translate clearly in the one-hour episode, but it's an important distinction I always remind myself of in watching their relationship unfold.
One of the single greatest moments in Trek hitory, a mythmaking scene, is when Edith sizes up Spock and Kirk together– Spock, always standing by Kirk's side and Kirk belonging off somewhere else. Joan Collins deserves credit for doing such a good job in a barely-scripted character. She plays Edith perfectly. And her brief assessment of the Kirk-Spock relationship makes Edith important too, it shows that she gets it, she understands these two men, the two central characters to Star Trek. In another reality, where Edith were allowed to live and be with Kirk, you could see her fitting into the Kirk-Spock dynamic seamlessly. That sort of thing puts Keeler over the top as a Kirk-girl. She is the product of excellent writing and very good acting.
People that criticize Shatner's emoting skills lack both good judgement and artistic awareness.
The play of features on Shatner's face after the trio emerge from the guardian of forever conveys more agony, torment, regret, and grief, than ANY thespian could impart with a 15 minute monologue, and SHatner didn't utter a word. His expression said everything you needed to know, followed by the immortal "Let's get the hell out of here", a verbal summation of what has transpired for those that need things spelled out for them.
Shatner is utterly UNDER-rated. A damn fine actor that understands nuance despite his reputation, and posesses all the creative attributes to completely embody a mythical fictional character.
The Shat is the man.
Jim J
I agree, Josh. Honestly, Shatner has proven himself beyond a doubt. How many actors his age are in as high of demand? It's obvious that people are finally starting to understand and appreciate "the man". Though I have always felt this episode is over-rated, I can certainly understand why people love it…this review totally captures why. An excellent review that is very accurate!
Dennis, kudos on a superb review. It will be a challenge for CBS to decide what to revisit and what to leave to our imaginations. I agree that the folks working on these editions should be acknowledged in the credits – they deserve the credit (and the blame) for what works and what doesn't. Here's hoping they work on improving the nacelle shots.
I'm not sure why all the complaints about the nacelle lighting effects. In my opinion these are fine, a nice "retro" effect very much like the original. What's the big problem here, I just don't see it.
Every time I see this episode, I think what a wasted chance there was in GENERATIONS to have Joan Collins do a cameo as Edith in the Nexus.
To be fair, GENERATIONS had a million missed opportunities. The TREK honchos should have shelved YESTERDAY'S ENTERPRISE and saved it as the script for the first film.
Just sayin'.
Re 8 ;Edith could mud wrestle Linda Evans in the nexus!
Yeah, when will they make "Dynasty: The Motion Picture?"
Honestly, I'd sooner they left this episode alone. Bringing Keeler back in any context would cheapen the story and character, IMAO.
Maybe they shouldn't have featured Kirk in "generations" at all, because it seems it was a gimmick and didn't service the character, but rather was a marketing ploy.
I think that's why Nimoy wouldn't commit to direct, the story had no substance.
Featuring Kirk was Berman's ill-concieved idea to benefit and support Picard, at the expense of Kirk.
The scene where Picard continually tries to coax Kirk into action is laughably bad. You can't tell me a character with the experiences Kirk has would be deluded into frying eggs several moments after a bulkhead blows away and he finds himself chopping wood. Come on now, this is Captain Kirk here. He would have immediately questioned his surroundings and enviroment, not immerse himself in the surreal experience.
"Generations" was bad. Very bad. I find it telling even Ron Moore and Braga later say the same thing.
DB …Yeah!Dynasty ;the Motion picture.original cast!
Perhaps I'll get a answer here? Does anyone know IF they plan on updating and showing the original pilot to Star Trek: The Cage?
Daren R. Dochterman
This has strayed off topic… but when someone brings up "Generations" it's a sore spot with me.
The Kirk Nexus setting was not the Kirk we know. This is the Fantasy of William Shatner, not Kirk. Shatner is the guy who would love to spend eternity chopping wood and riding horses and scrambling eggs. Kirk's true love was always the Enterprise. Kirk in the nexus would be at the helm of the TOS ship… exploring strange new worlds, and whompin' Klingons. I think if they had brought Kirk out of the nexus and had him at the helm of the "D" and it was him battling away and crashing the ship, that would have been a much better tribute to the character. Just my two cents.
Anthony: Thank You for your answer….I appreciate it! I posted it countless times and you finally answered it. Except I wonder this…I wonder why not?
Don't you….(and others) agree that this episode is worthy of a update too?
Perhaps the simple fact that it was a failed pilot is the answer in itself is why those at CBS aren't redoing it? As is…I been thinking of buying the entire episode….(it is available – right?) on DVD – since it certainly had its moments despite it all.
Right Daren,
Kirk was Han Soloed in Generations, (a reference to Ford's phoned in performance and Han's bad writing and characterization in ROTJ).
Shatner showed up for the paycheck, which I can't blame, but I do blame him permitting some of the horrible creative decisions he permitted as far as his character.
Which it seems he regrets now as well, frankly all parties involved seem to.
Killing off Captain Kirk is to Star Trek , as killing off Luke Skywalker is to Star Wars. It's silly, unnecessary, and effectively neuters your franchise.
It isn't dramatic. It isn't a reminder of life.
We watch Star Trek for escapism, not to be reminded of life. We watch it for the fictional universe where fictional characters do fictional things.
Berman buried Kirk because he wanted to bury TOS, it's as simple as that.
He touts Kirk coming back as giving him " one last bout of heroism."
Where is the heroism?
The initial draft had Kirk getting shot in the back like a **** and Berman was evidently alright with that. Only the test audience and their good judgement gave us the ending we DID get, which isn't much better but at least they tried to preserve SOME element of the character.
"They say time is the fire in which we burn."
Im sorry, but TIME sure as hell isn't kind to that film.
GENERATIONS is Star Trek at its worst.
I don't regard it as canon. In addition to Kirk's silly behavior/death, what about the destruction of the Enterprise-D? What a terrible fate, a weak ending for a noble ship. To be beat by Lursa and Betor? I am amazed at how poorly that show was written, even down to the Enterprise-B crew dialogue. Kirk, Scotty and Chekov were amusing, but that has more to do with their innate charm as actors than the words they were given. ALan Ruck was a terrible choice to be the Enerprise-B captain, so wimpy and unsure of himself. Demora Sulu was cute, I have to give her that, but man did that whole movie just stink.
By the way, as a total aide, I watched TMP again the other day after reading Christopher Bennet's novel "Ex Machina" and I have to tell you that movie has grown on me alot. If anyone trusts my opinion (which you have no reason to honestly) read the book and watch TMP again. The book puts meat on TMP's bones. And it's a sequel to TOS' "For The Earth Is Hollow And I Have Touched The Sky." Excellent stuff.
KAAAAAHHHHN
As bad as GENERATIONS was, the entire film would have been saved if, IF, in the scene where Kirk decides to go up the stairs to ask to "marry that woman," he would have found himself on the bridge of his old ship instead of that stupid horse barn after he stepped through the bedroom door. His true love was the TOS Enterprise, not some female character buried in subtext that we never met. But alas, our franchise had been officially hijacked by then, and I quickly abandoned Star Trek movies from that point on.
I had wondered about "The Cage" being upgraded as well but from what I found it looked like it would be done like the rest? Hmm, will have to see if I can find that again.
Wow not updating the Cage flies in the face of the whole idea of Trek Remastered are we certain of this Tony?
What needs a good restoration more then the oldest Trek around?
They are doing The Menagerie which is made of up of 2/3 to 3/4 Cage footage.
I liked "Generations" well enough, though it's not one of my favorite Trek movies. It's nowhere near the worst.
Kirk's "fantasy" had to be something not that he had experienced but something that he had thought in idle moments might have been the alternative to his life — same as Picard imagining himself a happily married, homebound father when every decision of his real life had taken him in a direction similar to Kirk's.
If Kirk's Nexus experience had in fact been himself commanding the Enterprise, a lot of people would have complained about his willingness to leave it.
One of the things that I like about "City On The Edge Of Forever" is that it presents Kirk as a human being with his own quirks and frailties, rather than the superhero that decades of fan adoration would insist upon. Super-Kirk is no more or less interesting than Johnny Weissmuller as "Tarzan" — entertaining within limits, but with circumscribed potential as a character in any kind of new drama or fresh setting.
One thing that I hope Abrams and his writers can accomplish is to portray pre-TOS Kirk as someone more akin to the character in episodes like "Balance Of Terror" or "Dagger Of The Mind" or "City On The Edge Of Forever" rather than continuing the near-parodic interpretation of Kirk in the later episodes of TOS and most of the Trek films.
I've always been a champion of Star Trek – The Motion Picture, Adam.
It was the Trek film I rented and watched over, and over, and over, and over when VCR's first came out that featured the whirring lift to insert the tapes on top, God that's old school, we are talking 1980.
I think the media has somehow propogated the notion The Motion Picture was substandard more than anything else.
I equate it to Wrath of Khan as far as being my favorite, though naturally the films themselves could not be further apart.
I think when you watch The Motion Picture, and The Wrath of Khan, you are essentially witnessing every aspect that Star Trek represents.
With the Motion Picture you get the epic space odyssey, pro-humanistic ideologies, high brow concepts, unknown life forms, old school production design and cinematography, and larger than life characterizations of heroic individuals.
With the Wrath of Khan, you get the character study, story arcs, black hat villains, relateable characterizations, humor, Literary allusions, and space battles.
Combined, you have every quintessential idea that Star Trek stands for.
I still maintain the Motion Picture is the most cinematic and "professional" of ANY Trek endeavor TV or film, it had Orson Welles narrating the trailers for Kirk's sake! That alone is a testament to it's epic scope.
Plus, never before, or never since, has the starship Enterprise looked so damn good!
I tend to agree with what Josh said.
For a long time I went with the mass opinon that TMP wasn't very good. But what's funny is that if it was on cable or I was bored I would often pop in the video tape and watch it. There is something compelling about the story. Sure it's a bit out of place with the rest of movies (which doesn't inherently make it "bad"). As a kid I always liked the faster paced TWOK-TSFS arc, yet I was still drawn to TMP, it drags on a bit I still think. Seeing the more complete Robert Wise vision really clinched it for me when the Directors Edition was issued, delved into the extras to see what was done and how the director really wanted to do more. The new CG wasn't too hot in some places even then, and certainly hasn't aged well in the High-Def era but that's not the point. In many ways TMP is more like a foreign film, more abstract.
And of course TWOK has some incredible action and eye for Naval-like details which makes it compeling. A spirtual successor to Balance of Terror in some ways. It has been noted repeatedly something Mayer did right in TWOK and later in TUC is make the ship full of people with jobs to do, people with tools in hand, manning stations, etc. Things like this were often seen in TOS, especially the first season. This, as Josh said, brings back the classic Hollywood formula of clear bad/good and even has the cliché happy ending in TSFS (completing the Genesis planet story arc).
Matt don't forget that guy with that floating slab of carbonite in the day-glow blue hallway that Kirk rushes past!
He was doing something , but the question is, what WAS that guy doing and where was he going with that thing anyway??
How about the dockworkers waving as the Enterprise departed? ;)
I must admit, I like and prefer the "The Motion Picture" Klingons to any incarnation we have seen, they truly looked alien with their swarthed hairstyles, and white contact lenses.
Every single time I see the first Klingon bridge view with that slow zoom on Mark Lenard and that Officer walks up to his shoulder, it literally looks and feels completely alien, with the subtle smoke, red lights, and awesome costumes and makeup.
It just has alien submarine written all over it.
But of course, it's short lived as they get annihilated like *******.
Oh, and they run too, how often do you hear or see of Klingons RUNNING?
Their was one book that was for sale at one time that showed the story boards – outlines and concepts of Star Trek TMP and even the 2nd series that never seen light of day on Tv….has anyone seen this book too?
It was very good in the ideas that it had and master illustrations. I do believe if I remember correctly reading that many of the concepts that they had were never realized simple due to lack of money (primarily) and time.
They showed the sickbay – a revamped bridge and much more. I know that many of you have seen the Enterprise as they envisioned it. The lower
back half was flat and looked like a aircraft carrier – etc. Well….anyhow!
mike g.
I am with those of you who feel "City" was the best episode of Star Trek. The original teleplay was even better than the filmed version, but the core and heart of the story still came through. I've always felt Harlan Ellison is one of the finest writers around, and he always managed to get a bum deal from non-creative executive-types. Now, this is a great segue into TMP, because Ellison had pitched an idea for that film, which many of you may already know. Essentially, it was another idea where Kirk had to make a decision- did he have the right to wipe out an entire species in order to save humanity? Making choices, making difficult decisions — this is the kind of story-telling which made for great tv-film, yet we rarely ever saw, especially in those later versions of Star Trek.
That said, I do enjoy ST-TMP, especially the revamped director's cut. Just goes to show how some very subtle things can make a huge difference in the big picture. What I appreciate about that film is its attempt to convey realism, and a sense of exploring the unknown. Every other Trek film – good or bad – was just a popcorn film.
My hopes for the new Trek film is that it rise above popcorn mentality, and give us something of real substance… the kind of substance we got to see in "City".
bdrcarter
OK…so back to the Remastered reviews. The more I watch of the new, old episodes, the more disappointed I become. I had high expectations after seeing the work by Daren Dochterman and Eden FX that cropped up shortly before Remastered premiered. CBS/Paramount missed a big opportunity by passing on those guys! I think the vision of their proposals was the right balance of respect for the original work and the possibilities of CGI. I wonder what Daren would have done with the decaying orbit in The Naked Now? At least one exterior shot of the ship heating up in the atmosphere, maybe? What could the Guardian's planet have looked like with a pan of the ruins on the surface? There was plenty of time to have a Kirk POV shot in the horta's nest that paid off the horror of millions of broken eggs instead of a small handful. These are a few examples of missed chances to expand on the original vision of Star Trek with today's technology without changing a beat of the storytelling. And the shots that have been recreated don't seem fully realized. I don't expect to see rivets on the ship's hull but where is the added depth we were hoping for? The closing shot of the Enterprise in The Naked Now looks like a plastic model kit. How about some subtle enhancements, like illumination from the impulse engines? I'm also surprised they haven't tried to enhance some of the other opticals. I assumed (incorrectely) that they would update the hand phaser effects. So far, the cartoony and inconsistent effects of the 60's live on. (I'm glad they added a beam to Scotty's phaser as he was cutting into the bulkhead..but where was the sound effect?) I do think the new planet visuals are amazing and there have been a few nice surprises, like the exterior of the science station in The Naked Time, the close ups on Spock's tricorder and the matte work for the mining facility in Devil in the Dark. (The opening shot with the digital guard seemlessly walking into the next shot in the cave?…Brilliant!) Unfortunately, that's the kind of work I was expecting to see all the time, so I can't help but feel disappointed. I think it's time for a visit to trekenhanced.com to see what could have been. I hope someone at CBS gives it another look too.
(BTW, totally agree on TMP being underrated and that Generations was so bad that it singlehandedly started the decline of the franchise.)
I agree that the nacelle complaints are unjustified. To me, they got the color and the effect just right. (see http://www.retroweb.com/trek/trektv8a.jpg for comparison)
If I had to complain, It would be that the inset spherical end caps (aft) on the nacelles are slightly too small, and they are not reflective as were the originals (see http://www.retroweb.com/trek/trektv2a.jpg )
If the links above don't work, copy and paste them into a new browser instance, and they should work.
By the way, have we see a view of the Enterprise underside yet? Does the CGI-generated Enterprise faithfully include the square-shaped red marking under the shuttlecraft bay? See http://www.retroweb.com/trek/trektv9a.jpg which is from The Alternative Factor (the only time this angle of the Enterprise was ever seen)
See, I don't think that the CBS Digital effect looks very much like that at all. It's much brighter — far too luminous compared to windows, the saucer domes and so forth — too color-saturated and the blades are too distinct. The whole thing looks more like cell animation than any kind of real lighting effect.
Rob Bonchune's effect for the Defiant in "In A Mirror, Darkly" was vastly superior.
Nice snapshot of Mc Coy.I especially like the glint of light off that one tooth.
Dennis Bailey
Revisiting this preparatory to writing another review, I noticed that I made a big error – I meant to describe the Guardian's Planet as rotating at something like three hundred miles a *minute* not three hundred miles an *hour* – the latter would be on the slow side.
Keeble Walker
James T Kirk: Is how "I' view you, watching you, over the years, when you shwo your self, acting as you have you, have left A, asking question two me, and all it was as my eye's seen it, all it was was A, "Airplane wing" your unheard voice ove saying its out their on that, "Airplane wing" as it was very harie and ugly, what is reall, and what is, fake, what we see and what we dont see, and all it is, is A, "LARGE FEET" which shows it self its feet never the face the feet shrinks are remains large, and yet why, you as you are is rellie know as, "William Shattner" my name and who "I' undress my self two each ove you over the years has ment nothing, as we can see, and yes James T Kirk um not A, M9 sayer, and yes who was Edith Keeler, as you can see my name is, Keeble, and yes who rellie was, "Edith Keeler" and why as it is said, "Edith Keeler must die" ive search aasking the, inter search enging, info baout Edith Keeler, and yes how are you, "James T Kirk, yes you, William Shattner" when, "I' watch you each time with in my, "Mines eye, "I' payutenchen" knwoing each faser word means somthing, like this, word, "InFractionInKrishun KripshunInfractionFraction=IN with in the mouth is the, Infraction
David Avery
This is perhaps the best of all the episodes from the famed "Star Trek" series from the 60's. My father would let me stay up late to watch them when I was much younger. The point where Kirk holds back Macoy from saving Edith Keiler from being killed was the best acting scene for all three actors. The cold "must be done" stare of Spock; the "frantic" attempt at saving her by Macoy; and the "heart-breaking" reaction of what just happed by Kirk. One would think he would of decked Macoy for his statement, "Jim…do you realize what you just did?", followed but Spocks' s response of, "Yes Doctor…he does."
Locutus
Changing *anything* to Star Trek episodes is heresy and abomination ! They shouldn't have changed a single frame of the original print, like you don't CGI "enhance" a painting or statue. It's a work of ART, it is what it is, if you change it you damage it by definition.
Shame on whoever had the dirty job of defacing Star Trek episodes, THEY must have insisted themselves NOT to be credited and go down in Trek history as the vandals of TOS !!
Solar Trekkie
There were only four things good about Generations. The rest was just "you-know-what."
1. TOS first half
2. Malcolm McDowell(the last person I would ever imagine being on ST
3. Lursa and B'Tor being killed off.
4. Whoopi Goldberg, John Putch, Tim Russ, and Jeanette Goldstein


text:
Gambia must heed UN warning to improve its human rights record
28 October 2014, 00:00 UTC
The Gambian authorities must heed a warning from the international community about the deteriorating human rights situation in the country, Amnesty International said today.
This morning, 62 countries took the floor at the United Nations Universal Periodic Review (UPR) in Geneva to urge Gambia to promote and protect human rights. Concerns voiced by UN member states included Gambia's unjustified restrictions on the right to freedom of expression and its renewed use of the death penalty.
"UN member states have sent a clear message to Gambia that the government must end its rule of fear and repression," said Steve Cockburn, Amnesty International's regional deputy director for West and Central Africa.
"Gambia's muzzling of dissent has had a devastating and chilling effect on human rights defenders, journalists and political activists, who have been persistently brutally targeted solely for exercising their right to freedom of expression. Today UN member states have said to Gambia: enough is enough."
Journalists in Gambia have been subjected to enforced disappearance, while activists have faced torture – including having their legs burnt with boiling oil and their backs slashed with metal rods – for speaking out against human rights violations by the state.
Member states also criticized the Gambian government's disregard for regional court judgments and the resolutions of international and regional human rights bodies.
"It is disappointing that the Gambian delegation in Geneva repeatedly sought to play down the grave and increasingly deteriorating human rights situation in the country," said Stephen Cockburn.
"Gambian human rights defenders experience a very different reality than that portrayed by the delegation to the UPR Working Group."
In blatant contradiction to Gambia's obligations under international human rights law, Gambia's Minister of Justice said restrictions on the right to freedom of expression were necessary to avoid the publication of false news, sedition and libel.
Gambia justified its use of the death penalty and argued that the temporary lifting of the moratorium on the death penalty had been necessary to deal with a rise in crime.
Gambia's constitution in fact calls for a review – now seven years overdue – on the "desirability" of the death penalty.
"The climate of fear and intimidation in Gambia has affected the ability of civil society to participate in this UPR and there is a real risk of reprisals against Gambians who sought to engage with the review," said Steve Cockburn.
"UN member states have today echoed what civil society has been campaigning on for years. Gambia must accept the recommendations made during this session and genuinely commit to fully and effectively implementing them without delay."


text:
Boadicea's Chariot
Hic Leges Icenorum Observantur
Who can do what?
Basic Resizing
Fun Factoids
This is the way your world ends…with a bang
Just finished reading Crime and Punishment for the second time. The first time was twenty years ago when it was all Russian to me. Being older I had a better understanding of the novel this time.
One of the passages that caught my eye on this reading was when Raskolnikov, after five days of delirium, reads the previous five days newspapers. He is searching for details of the double murder. Before he sees the reports of his crime, there is a flavour of the news in Petersburg of that era. There is an accident on a staircase, four incidents of fires (was there an arsonist on the loose?) and spontaneous combustion of an alcoholic shopkeeper. This brought a smile to my face as I thought of a funny episode in a film.
In a scene from the rock music spoof, This is Spinal Tap, one of the band's drummers dies from spontaneous human combustion. Most of the drummers, party animals, seem to have a small shelf life and die from various causes. But if you gotta go. SHC is as good as any.
Author theroyalistPosted on February 13, 2016 February 13, 2016 Categories General, Terminally boring, The Dark Side, UncategorisableTags Crime and punishment, drummers
13 thoughts on "This is the way your world ends…with a bang"
janus says:
J-Dubya, if he'd been a rodeo rider, would he have got more bangs for his buck?
Araminta says:
Blimey, JW. SHC sounds a bit, well messy!
Sipu says:
Bleak House, written in 1852-3 also features SHC. It was for me, an extremely disappointing aspect of that great novel.
Dickens also quotes other 'examples' of SHC. Since naked flames were common in Victorian times, I wonder why he bothered?
jazz606 says:
My reading of War and Peace is deferred because I'm reading The Silk Roads by Peter Frankopan and at the same time(ish) Chusan by Liam d'Arcy-Brown . Although about different parts of history they're are largely about trade and the wars concomitant with it. In The Silk Roads one learns that religion is also (mainly ?) about war and trade.
As far as War and Peace is concerned I have recorded the BBC series and I guess I'll eventually bring myself to watch it.
theroyalist says:
Afternoon Jazz,
It took me a month to finish War and Peace. Two previous attempts got me halfway through but thanks to the nudge by the TV series I made it to the end this time. The last few chapters (the epilogues) were tough going; very repetitive. After this I took another stab (no pun intended) at C&P. Having had enough of Russian literature for the time being and looking for something different a friend recommended the works of David Mitchell -The author not the "comedian". I've almost finished Ghostwritten, his first book.
I am reading Barchester Towers, published in 1857, a year after C&P and 12 years before W&P. Having read the Russian novels I can assure you that English one is much less heavy going.
christinaosborne says:
I am reading, or trying to read-
The Isis Apocalypse, the history, strategy and doomsday vision of the Islamic state.
By a William McCants A fellow of and a director of the Brookings institution.
My God what a carry on. All the different branches of Islamic uprising fighting like cats in a sack. Reading between the lines the worse thing we ever did was to bump off Bin laden when we did, serious mistake! His was the most moderate and ameliorating voice there was!!!! (By then)
And then you get to their apocalyptic visions, which is what they are really fighting about, makes the Book of Revelations look reasonable!
It becomes totally apparent that it is going to be a game of 'whackamole'. One Islamic state pops up, you smack it down and the nutter survivors take themselves off to another muslim country and off it goes again!
They have done it now and failed several times. Mali, Yemen, Algeria etc, now it is Syria and Libya's turn.
Just Wait till they get to Birmingham.
One gets the ghastly impression that this turns into a Thirty year war that we speak of so glibly in a historical reference. Seriously not good. Especially if we allow the whole thing to become WWIII by proxy.
The ultimate cluster fuck to destroy our world over a bunch or mad rag heads.
Makes your SHC look like a walk in the park. Very depressing bedtime reading I do assure you makes Crime and Punishment look like a comedy!
Get back to Barchester Towers, CO – easier on the hackles! 😊
Game of Thrones has a pretty good take on current events.
Have read and reread Thomas Hardy n times already. Not again please!
I must admit to a predilection to expert's books however unreadable. I find the media are so thick or they think we are so thick, or both, that their background explanations are so inadequate as to be laughable.
Know one's enemy, so few do. Had the British studied the Sino Soviet war of 04-05, Gallipoli and the massacre of Cavalry regiments in France would NEVER have happened. The sick making thing was that the British, were there as observers!!! Note, the Germans had tooled up the Japs with God knows what armaments which were all prototypes for 14-18 and they thrashed the Russians something rotten who fought a Napoleonic style engagement. Another nice little proxy war that was for the main engagement.
sipu I don't do fantasy, the past is quite bad enough!
Hi CO, sadly, I don't do history any more. It is so perverted by modern historians to fit the politically correct agenda, that I do not believe anything I see or read that was not written 50 years or more ago. At least with fantasy there is no attempt to present it as anything more than that.
Not sure about the reference to Hardy. Barchester is Trollope.
I am not sure that military commanders and their political masters ever learn very much from history, military or otherwise. My grandfather fought in the Boer War as one of the original British commandos. This small unit, the Lovat Scouts, comprised Highland men who were the first to wear proper camouflage, based on ghillie suits. The unit had been put together to beat the Boers at their own game. Interestingly they were commanded by the American Frank Burnham who had fought in Rhodesia in the Matabele War and taught Baden Powell all he knew about Scouting.
In any event, one would think that the British commanders would have learned from the experience of fighting the Boers how not conduct warfare in a hostile territory. Clearly not. My grandfather went on to fight at Gallipoli where he was wounded.
Oh heavens your right! Wrong author! Oh the brain is going gaga.
Will write about my father and Gallipoli tomorrow.
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Today is a SNOW day!!!
even if it's only for the new challenge at The Ink Spot!
I do miss my winter and snow very much!
Have you heard the news about the monthly stamp kits from The Ink Spot?
stamp kit you can sign up for and it's delivered monthly, right to your mailbox!
Today is the day that we Dueling Darling post our cards.
a card kit exchange friendship group.
Be a Kid at Heart!
matching fun image and sparkly sequins.
but I can definitely share my card with a child ~ my own!
have created with 1 theme, 2 different card kits!
February is so much fun!
surely has the right idea with 'tagging' the fence.
we read "Harold and the Purple Crayon" together.
that we did a recreation on his own bedroom wall!
and our girls did a trio of "Hey! Why didn't WE get to do that!"
It's post day for the 3 Musketeers!
because I'm running everywhere else - LOL!
He has his school's Principals' Honor Roll Assembly today.
I told my husband 'Look! It's me on a card!"
he offered "Baby, that is definitely not you on the card."
Well, I looked again and lo and behold, he's right!
I sadly do not own those cool heart shaped glasses!
with all the little handwritten To and From names and notes!
I think the kids should make them!
"It's all about the candy, mommy!"
How about Blue! Orange! Teal!
(Nope, not channeling the Broncos for a win - LOL!
Stamping Riley is getting ready to send some love!
adding him to a colorful palette of blues and orange ..
by checking out the blog/links here!


text:
APPEAL DENIED: Josh Gordon Suspended One Season
All Videos, Featured, NFL News
According to NFL Media Insider Ian Rapoport, the NFL has suspended Cleveland Browns wide receiver Josh Gordon for one season for violating the league's substance-abuse policy. Gordon lost his appeal and is now scheduled to miss the entire season as there is now no chance for him to return.
Gordon and his camp participated in an appeal session in front of arbitrator Harold Henderson earlier this month and he felt he has posed some serious issues that would lead to his suspension being reduced if not lifted entirely. Gordon was wrong on both counts.
Steven's Take:
There goes the Browns offense, literally. Gordon WAS the offense for the Browns last season. In just 14 games last year, Gordon hauled in 87 passes for 1,646. That was over 40 percent of the Browns offense through the air. Over 40 percent!
His absence is going to be sorely missed and regardless of it is Brian Hoyer or Johnny Manziel under center, the Browns are not going have the same aerial attach from a season ago. They will now need to rely on the running skills of Ben Tate and rookie Terrence West. Their best threat in the passing game now is tight end Jordan Cameron.
3 Responses to "APPEAL DENIED: Josh Gordon Suspended One Season"
How can Josh Gordon get at least 12 months and Ray Rice get 2 games? It is a shocking endictment of the lack of transparency shown by the NFL. This decision reflects poorly on league and those at a high level making the choices that guide the course and future of the game we all love. Disgraceful!
The Commish and the league answered you today. They know they made a mistake with the Ray Rice situation and now a player will get 6 games for 1st offense and be banned for life for 2nd offense. A little late, but step in the right direction.
Steven, thanks for the reply. Notwithstanding the leagues changes announced today, the offences committed by Messrs Gordon and Rice are at very different ends of the criminal spectrum (being a Brit I not entirely familiar with the State/Federal criminal justice system) and the punishments doled out to them do not appear to be representive of the public feeling on the unlawful acts of those men or bear much, if any, correlation to the non-NFL sanctions either could receive.
That being said a published tariff of penalties for players and to bind Goodell can only be a good thing.
In summary: Gordon stupid but unlucky; Rice stupid and very lucky. Transparency and openness better.