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US 7279292 B2 Abstract A method for identifying low grade astrocytoma cells in a sample is provided, wherein the method distinguishes between low grade astrocytoma cells and normal astrocytes, thus permitting early diagnosis of astrocytoma. The method uses antibody directed against J1-31 to test astrocytes in the sample for the presence or absence of J1-31 polypeptide, the low grade astrocytoma cells being characterized by the absence of J1-31 while normal astrocytes are characterized by the presence of J1-31.
Claims(11) 1. A method for identifying low grade astrocytoma cells in a biological sample, comprising the steps of:
preparing slides from a biological sample;
serially incubating said biological sample in at least ethanol and hydrogen peroxide;
heating said slides in an unmasking solution including a mixture of at least sodium citrate and citric acid;
incubating the biological sample with anti-J1-31 antibody under conditions in which the antibody binds to J1-31 antigen in the sample, said anti-J1-31 antibody being produced by the hybridoma cell line J1-31, ATCC designation number CRL-2253;
contacting the biological sample with a secondary antibody selected so as to form an immune complex with the anti-J1-31 antibody bound to J1-31 antigen;
detecting cells expressing the J1-31 antigen by detecting the immune complex; and detecting astrocytes in the sample in which the immune complex is not present, to thereby identify low-grade astrocytoma cells.
2. The method of
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incubating the immune complex with a conjugate; and
exposing the conjugated immune complex with a chromogen.
Description This application is a divisional application of U.S. patent application Ser. No. 10/269,773 filed Oct. 10, 2002, now abandoned which claims the benefit of U.S. Provisional Application Ser. No. 60/328,917, filed Oct. 11, 2001, the content of which is hereby incorporated by reference in its entirety. The subject of the present invention is a method for using an antibody to detect the presence of a disease, and more specifically to distinguish low grade astrocytomas from normal reactive astrocytes. Astrocytomas are members of the glioma family of tumors; that is, tumors arising from glial cells. In the case of astrocytomas, the tumors arise from a type of glial cell called astrocytes. Astrocytes, named by Spanish neuroanatomist Santiago Ramon y Cajal in 1913 after he discovered the star-shaped cells, are the major cell types of the central nervous system that responds to various pathological conditions, including trauma, ischemia, demylination, inflammation, etc. In response to such pathological conditions, the normally quiescent astrocyte becomes "reactive," and may proliferate and migrate, exhibiting hypertrophy with increased expression of Glial Fibrillary Acidic Protein (GFAP), expression of cell surface MHC class I and II molecules, and producing various cytokines and growth factors. Neoplastic transformation of astrocytes give rise to a variety of astrocytomas. Astrocytomas are generally grouped into two categories: low-grade (grades 1 and 2) and high-grade (grades 3 and 4). High-grade tumors grow rapidly and can easily spread throughout the brain, while low-grade astrocytomas are usually much more localized and grow slowly over a longer period of time. The prognosis for patients with high-grade astrocytomas is poor, with an average survival of 18 months for patients with grade 3 astrocytoma (with radiation therapy and chemotherapy) and an average survival of from 17 weeks (no treatment) to 51 weeks (with surgery and radiation treatment) for grade 4 astrocytomas. Morphologically, low-grade astrocytomas are usually normal in appearance, making it difficult to distinguish low-grade astrocytoma cells from normal quiescent or reactive cells. Some grade 2 low-grade astrocytomas may be slightly abnormal in appearance, but are not easily detected by routine microscopic evaluation of tissue biopsy samples. By contrast, the high-grade astrocytomas are abnormal in appearance and show evidence of mitosis, making the cells easily identifiable by microscopic evaluation. Visualization of astrocytes during microscopic evaluation of tissue sections is typically facilitated by staining the biopsy tissue sample with antibody to GFAP, a cytoskeletal protein expressed both in quiescent and reactive astrocytes and in low grade and high grade astrocytomas. Both monoclonal mouse and polyclonal rabbit anti-human GFAP antibodies have been developed that are specific against GFAP, not recognizing other intermediate filament proteins. These antibodies typically react with both astrocytes and astrocytoma cells, and thus do not serve to differentiate between normal and transformed cells beyond facilitating morphological inspection Although the prognosis for patients with grade 1 astrocytoma is good, with some patients known to live 30 years or more following diagnosis, the prognosis for patients with grade 2 low-grade astrocytoma is much less optimistic. Recent studies have indicated that the 5-year survival rate in grade 2 astrocytomas is about 34%. With radiation therapy, the 5-year survival rate increased to about 70%. It is apparent that early diagnosis, and thus early treatment, is important in treating patients with low-grade astrocytoma. What is needed, therefore, is a reliable and efficient diagnostic procedure for differentiating low grade astrocytoma cells from active and quiescent normal astrocytes in tissue biopsies. One aspect of the present invention is a method for identifying low grade astrocytoma cells in a sample containing astrocytes, comprising testing the astrocytes in the sample for the presence or absence of J1-31 polypeptide, the low grade astrocytoma cells being characterized by the absence of J1-31. Another aspect of the present invention is a method for distinguishing low grade astrocytoma cells from normal astrocytes, the method comprising the steps of:
Another aspect of the present invention is a method for identifying low grade astrocytoma cells in a sample, comprising the steps of:
Another aspect of the present invention is a kit for in vitro diagnosis of low grade astrocytoma comprising anti-J1-31 antibody, a medium suitable for formation of an antigen-antibody complex, and reagents for detection of the antigen-antibody complex. Another aspect of the present invention is a kit for in vitro diagnosis of low grade astrocytoma comprising anti-J1-31 antibody and instructions for using anti-J1-31 antibody to distinguish between low grade astrocytoma cells and other astrocytes. These and other features, aspects and advantages of the present invention will become better understood with reference to the following description, appended claims, and accompanying drawings wherein: The present invention relates to methods of identifying low grade astrocytoma cells and distinguishing such cells from normal, reactive astrocytes, using anti-J1-31 antibody. One aspect of the method of the instant invention for identifying low grade astrocytomas comprises immunostaining tissue biopsy samples with anti-J1-31 antibody to differentiate between normal, reactive astrocytes and low grade astrocytoma cells. Another aspect of the method of the instant invention for identifying low grade astrocytomas comprises immunolabeling tissue biopsy samples with anti-J1-31 antibody for fluorescence microcopy to differentiate between normal, reactive astrocytes and low grade astrocytoma cells. Still another aspect of the method of the instant invention for identifying low grade astrocytomas comprises subjecting tissue biopsy samples to immunoelectron microscopy using anti-J1-31 antibody to differentiate between normal, reactive astrocytes and low grade astrocytoma cells. Still another aspect of the method of the instant invention for identifying low grade astrocytoma cells comprises extracting proteins from tissue biopsy samples and immunoblotting with anti-J1-31 antibody to differentiate between normal, reactive astrocytes and low grade astrocytoma cells. Still another aspect of the method of the instant invention comprises testing cultured cells for the presence of low grade astrocytoma cells using anti-J1-31 antibody. Still another aspect of the present invention comprises kits useful for diagnosing low grade astrocytomas. J1-31. J1-31 is a cytoskeleton-associated polypeptide, which has been shown to be a 30-kD protein, distinct from GFAP and vimentin proteins also found in glial cells. By contrast, GFAP is a major cytoskeleton protein. The J1-31 polypeptide is expressed in both high grade astrocytoma cells and normal astrocytes, with enhanced expression demonstrated in reactive astrocytes following injury to the central nervous system (CNS). See, Singh, et al., (1986) Bioscience Reports 6: 73-80; Predy, et al., (1987) Bioscience Reports 7: 491-502; Predy, et al., (1988) J. of Neuroscience Research 19: 397-404; Malhotra, et al., (1989) J. of Neuroscience Research 22: 36-49; Singh, et al., (1992) Dendron 1: 91-108; Malhotra, et al., (1993) Brain Research Bulletin 30: 395-404; and Singh, et al., (1994) Biomedical Letters 50: 163-172, all hereby incorporated by reference in their entireties. Surprisingly, however, the present invention demonstrates that J1-31 does not appear to be expressed in low grade astrocytoma cells. Thus, J1-31 is useful as a diagnostic tool for distinguishing low grade astrocytoma cells from quiescent and reactive astrocytes, as well as from high grade astrocytomas. Antibodies. The present invention contemplates the use of polyclonal and monoclonal antibodies, including recombinant single-chain or other antibody derivatives or fragments, against J1-31 polypeptide and fragments thereof. The present invention also contemplates the use of polyclonal and monoclonal antibodies against variants of J1-31 polypeptides, including naturally occurring allelic variants. As used herein, unless specified to the contrary, "anti-J1-31 antibody" shall refer to a polyclonal or monoclonal antibody, including recombinant single-chain or other antibody derivatives or fragments, directed against J1-31 polypeptide(s) or fragments or variants thereof. Methods of producing polyclonal antibodies are well known to those skilled in the art. Exemplary protocols which may be used are described for example in Coligan et al., CURRENT PROTOCOLS IN IMMUNOLOGY, (John Wiley & Sons, Inc, 1991) which is incorporated herein by reference, and Ausubel et al., (1994-1998, supra), in particular Section III of Chapter 11. Alternatively, monoclonal antibodies may be produced using the standard method as, for example, described in an article by Kohler and Milstein (1975, Nature 256, 495-497) which is herein incorporated by reference. Anti-J1-31 Monoclonal Antibody. One example of an anti-J1-31 monoclonal antibody was produced using tissue homogenate from the brain of a multiple sclerosis patient and hybridoma technology, as reported by S. K. Malhotra, et al., (1984) in Microbios Letters 26: 151-157, hereby incorporated by reference in its entirety. This anti-J1-31 monoclonal antibody is specific for astrocytes but recognizes different antigenic site(s) than does anti-GFAP antibody. Specifically, this anti-J1-31 monoclonal antibody reacts with the astrocytic cytoskeleton. The J1-31 hybridoma cell line has been deposited with the ATCC as CRL-2253. Detection of Low Grade Astrocytoma Cells. As discussed above, it has been previously demonstrated that J1-31 polypeptide is expressed in normal astrocytes and in high grade astrocytomas, and may be detected in those cells using an anti-J1-31 monoclonal antibody. The present inventors have now found that J1-31 polypeptides are not detected in low grade astrocytomas, providing a new diagnostic method for identifying low grade astrocytoma cells, and thus facilitating earlier diagnosis and treatment of patients with low grade astrocytoma. In the practice of this invention, astrocytes are tested using anti-J1-31 antibody, as described below. Optionally, the cells are also stained with a non-cell specific stain, as with Harris hematoxylin, for example, which stains cell nuclei a deep blue color, allowing all cells to be easily visualized. Optionally, cells may also be stained with anti-GFAP antibody, to facilitate characterization of the morphology of the astrocyte/astrocytoma. Following testing with the anti-J1-31 antibody, the astrocytes are examined for the presence or absence of the J1-31 antigen and are identified, as summarized below in Table 1.
Thus, cells that appear as relatively morphologically normal astrocytes, but which do not contain J1-31 antigen, are identified as low grade astrocytoma cells. Methods for Detecting J1-31 in Cells. As will be appreciated by the skilled practitioner, there are several ways known in the art for detecting the presence of a specific polypeptide in a cell. Some of these methods are discussed below. However, the present invention contemplates any technique whereby the presence or absence of J1-31 in a cell, either cultured or in a tissue sample, may be determined. Immunohistochemical Staining of Tissue Sample Screening techniques such as Magnetic Resonance Imaging (MRI) are used to identify potential brain tumor sites, and biopsy tissue, as from a needle biopsy, is extracted from patients for analysis. Such clinically obtained tissue samples are generally formalin—fixed and paraffin—embedded, although tissue samples may also be fresh/frozen. Both are suitable for use in the present invention for immunohistochemical staining using an antibody directed to J1-31 protein. The tissue samples are then examined microscopically for the presence or absence of the J1-31 antigen in cells of interest. The procedure for preparing and staining these tissue samples is described below in detail. Prior to immunohistochemical staining, slides containing the formalin-fixed/paraffin-embedded tissue biopsy samples are warmed overnight at 60° C. Slides containing fixed tissue are incubated twice for 5 minutes each in Histoclear, a clearing agent available from National Diagnostics, Atlanta, Ga., followed by two incubations of 3 minutes each in absolute ethanol and two incubations of 3 minutes each in 95% ethanol. Slides containing formalin-fixed tissue samples that have been treated as described above with Histoclear and ethanol, are incubated for 20 minutes in a mixture of hydrogen peroxide and methanol. One suitable H2O2:methanol mixture is formed by adding 50 ml of 3% H2O2 to 200 ml methanol. The slides are next washed for 5 minutes in Delbecco phosphate buffered saline (DPBS). A 10× stock of DPBS is made by combining 11.5 g dibasic sodium phosphate (molecular weight [m.w.] 142.0), 2.0 g monobasic potassium phosphate (m.w. 136.1), 2.0 g potassium chloride (m.w. 76.6) and 80.0 g sodium chloride (m.w. 58.5), and adding dH2O to a final volume of 1 liter. A 1× working stock of DPBS is prepared by mixing 100 ml 10×DPBS with 900 ml dH2O. Following the DPBS wash, slides are placed in an antigen unmasking solution (0.1 M sodium citrate, pH 6.0), placed in a pressure-cooker, and heated in a microwave oven at 600 W twice for 15 minutes each. The antigen unmasking solution is made by mixing 810 ml of 0.1 M sodium citrate·2H2O (m.w. 294.12; 29.4 g/1000 ml) with 190 ml of 0.1 M citric acid·H2O (m.w. 210.14; 5.252 g/250 ml), for a final volume of 1 liter. Following the microwave treatment, the slides are left in the antigen unmasking solution for an additional 20 minutes at ambient temperature. Prior to immunohistochemical staining of freshly frozen tissue biopsy sections on glass slide, the fresh frozen biopsy sections are fixed in cold-acetone for 5 minutes, air-dried for 30 minutes and incubated twice in DPBS for 5 minutes each. The slides containing formalin-fixed or acetone-fixed tissue sections are next incubated for 20 minutes in a 1:20 dilution of non-immune horse serum to prevent nonspecific binding, (Cat. No. S-2000), available from Vector Laboratories, Burlingame, Calif. This is followed by incubation in appropriate dilutions of the anti-J1-31 mouse monoclonal antibody from 60 minutes to overnight. The appropriate dilution of the antibody is dependent on a number of factors, including the concentration of the antibody being used, and may be determined empirically by techniques known to one skilled in the art. Following incubation with the primary antibody, the slides are washed in DPBS four times, for two minutes each wash. The slides are then incubated in appropriate dilutions of biotinylated antibody for 30 minutes. One biotinylated antibody suitable for use with the anti-J1-31 mouse monoclonal antibody is a 1:200 dilution of biotinylated horse anti-mouse polyclonal antibodies (IgG, H+L), available from Vector Laboratories, Cat. No. BA-2000. This is followed by another series of four 2-minute washes in DPBS. The slides are next incubated for 30 minutes in avidin-biotin-horseradish peroxidase complex (ABC), available in a kit from Vector Laboratories, Cat. No. PK-6100. In this procedure, a preformed Avidin (A):Biotinylated enzyme (B) complex is used with biotinylated antibodies. 25 μl each of the "A" and "B" reagents provided in the Vector kit are added to 2.45 ml DPBS for a final volume of 2.5 ml. Following incubation, the slides are washed 4 times for 2 minutes each in DPBS. The final steps in the immunostaining procedure is to expose the slides with bound antibody-ABC to a chromogen substrate for the ABC enzyme. One chromogen suitable for use with horseradish peroxidase is diaminobenzidine (DAB). Finally, the slides are incubated two-times in 95% ethanol for 3 minutes each incubation, followed by two 3-minute incubations in absolute alcohol, and ending with two 3-minute incubations in xylene. The slides are then provided with a coverslip in permount mounting medium. Although one particular protocol has been described for use in immunohistochemical staining tissue samples with the anti-J1-31 mouse monoclonal antibody, other anti-J1-31 antibodies and immunostaining techniques may also be employed. For example, in addition to peroxidase conjugates, alkaline phosphatase conjugates may be used. Other suitable immunohistochemical procedures are apparent to the skilled practitioner. Immunolabeling for Fluorescence Microcopy Fluorescence microscopy may be used to detect J1-31 in astrocytes or high grade astrocytomas in cell culture or in tissue sections (Singh, supra, Dendron 1: 91-108). Briefly, for cultured cells, coverslips bearing cells are fixed in either methanol (5 minutes at −20° C.) or 4% paraformaldehyde (30 minutes at room temperature). The coverslips are then washed in phosphate buffered saline (PBS), pH 7.4, containing 0.05% Tween-20 (PBS-T). The coverslips with either an astrocytoma tissue section or cell line are incubated in 30% normal goat serum in PBS for 30 minutes to prevent non-specific binding. Anti-J1-31 antibody is added as the primary antibody and incubated overnight at 4° C. For example, antibody J1-31 ascites fluid may be applied at a 1:500 dilution in PBS; also, normal mouse serum (NMS; Sigma) may be applied to some coverslips as a control. Following a wash in PBS-T, an appropriate secondary antibody (for example, a 1:100 dilution in PBS of goat anti-mouse IgG conjugated to fluorescein isothiocyanate (FITC; Sigma)) is added and the coverslips incubated for one hour at room temperature in the dark. Optionally, the cells are double-labeled by first washing the coverslips again in PBS-T, followed by the application of a second primary antibody and incubation overnight at 4° C. Suitable antibodies for double-labeling included rabbit anti-cow glial fibrillary acidic protein (GFAP; Dimension Laboratories; 1:500 dilution in PBS), and its control, normal rabbit serum (NRS; Sigma); alternatively, Mab Vim 13.2 (a monoclonal antibody to vimentin; Sigma) and its NMS control may be used, also at a 1:500 dilution. The coverslips are washed in PBS-T and goat anti-rabbit IgG-TRITC (Sigma) or goat anti-mouse IgM-TRITC (Sigma) is applied at a 1:100 dilution in PBS. The coverslips are then incubated in the dark for 1 hour at room temperature Following incubation, the coverslips are given a final wash in PBS-T, mounted in glycerol/□-phenylenediamine and viewed by fluorescence microscopy and laser scanning confocal microscopy. Those cells expressing J1-31 antigen, and thus incorporating anti-J1-31 antibody and FITC-labeled secondary antibody, will fluoresce at a particular wavelength when excited, while any low grade astrocytoma cells present in the sample will not fluoresce at that wavelength, as no anti-J1-31 antibody/FITC-labeled secondary antibody is incorporated, allowing the low grade astrocytoma cells to be distinguished from the J1-31 containing normal astrocytes and high grade astrocytomas. Where double labeling is used, the second fluorescent dye is selected so as to fluoresce at a different wavelength (or in response to a different excitation wavelength). For immunofluorescence microscopy of tissue sections, the tissue sections are fixed in either 4% paraformaldehyde or methanol and sectioned using a cryostat or vibratome. Imprints on coverslips from fresh specimens may also be similarly fixed. Sections and imprints are immunostained as described above. Other fluorescent dyes, known in the art, may also be used in the practice of the present invention. Immunoelectron Microscopy Immunoelectron microscopy can also be used to detect GFAP and J1-31 proteins at the ultrastructural level in biopsy samples (Singh, supra, Dendron 1: 91-108). Briefly, specimens are fixed in 1.25% glutaraldehyde and embedded in LR white resin by routine methods. Ultrathin sections are etched with sodium metaperiodate and double-immunolabeled with anti-J1-31 antibody (preferably, monoclonal anti-J1-31 antibody) and anti-GFAP antibody as primary antibodies. The presence of J1-31 and/or GFAP is detected by the presence of the primary antibodies, as determined using secondary antibodies labeled with gold particles of distinct size (for example, 5 nm for anti-J1-31 antibody and 10 nm for anti-GFAP antibody). Thus, cells displaying both 5 nm and 10 nm particles (those expressing both J1-31 and GFAP; i.e., normal astrocytes and high grade astrocytomas) may be distinguished from astrocytes displaying only 10 nm particles (those expressing GFAP but not J1-31; i.e., low grade astrocytoma cells). SDS-PAGE and Immunoblotting The presence (or absence) of J1-31 in cells, both cultured and in tissue specimens, may be evaluated using standard sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blot protocols, which also allow the determination of the molecular weight of J1-31 protein (Singh, supra, Dendron 1: 91-108). Protein extracts from cultured cells (or tissue homogenates) are electrophoresed under reducing conditions in a discontinuous gel, then transferred to nitrocellulose or other suitable membrane by electrophoretic or other transfer. The membranes are then treated with anti-J1-31 primary antibodies and labeled secondary antibodies, using standard western blot procedures. In one embodiment, suspected astrocytoma cells can be selectively dissected from tissue samples using, for example, the laser microdissection technique of PALM® (P.A.L.M. Microlaser Technologies AG, Germany). Protein extracted from these cells can then be subjected to SDS-PAGE/western analysis to evaluate the presence or absence of J1-31. Kits for the Detection of J1-31 For diagnosis of low grade astrocytomas, a kit for the detection of J1-31 in tissue specimens and/or cultured cells is used. The kit comprises anti-J1-31 antibody, optionally J1-31 antigen for use as a control, and a means for detecting the complexing of the antibody with J1-31 antigen in the cultured cells or tissue specimen. The kit detects the J1-31 antigen, when present, with the anti-J1-31 antibody. The complexing immunoreaction is detected by staining, fluorescence, immunoprecipitation or by any other means used in the art and suitable for these purposes. In addition to the above, the kits may also comprise control compounds, anti-antibodies, protein A/G, anti-GFAP antibodies and the like for use in double-labeling, etc., suitable for conducting the different assays referred to above. Having generally described the invention, the same will be more readily understood through reference to the following example, which is provided by way of illustration, and is not intended to be limiting of the present invention. Formalin-fixed and paraffin-embedded tissue sections of a low grade astrocytoma were prepared and stained, as described above. Briefly, the tissue sections were treated as follows:
The slides were then prepared in permount mounting medium and examined microscopically at 200× magnification. Photomicrographs of the stained cells are shown in Cells stained with anti-J1-31 antibody are shown in The low grade (grade 2) astrocytoma cells, however, show no reactivity with anti-J1-31 antibody (note the absence of brown staining, for example, near the arrow, Thus, as demonstrated above, anti-J1-31 antibody can be used to quickly and efficiently diagnose the presence of low grade astrocytoma cells in a patient, allowing for improved screening and treatment of patients experiencing neurological cancer. All patents, patent applications, and other publications mentioned in this specification are incorporated herein in their entireties by reference. While this invention has been described in detail with reference to a certain embodiments, it should be appreciated that the present invention is not limited to those precise embodiments. Rather, in view of the present disclosure which describes the current best mode for practicing the invention, many modifications and variations would present themselves to those of skill in the art without departing from the scope and spirit of this invention. The scope of the invention is, therefore, indicated by the following claims rather than by the foregoing description. All changes, modifications, and variations coming within the meaning and range of equivalency of the claims are to be considered within their scope. Patent Citations
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These expression are so cute!!!
I say, excelente work, my friend!
You made Ratchet so cute in anime/manga of your style!!
My favourites are poker face, smiling, enrage, weeping, bashful (best favourite), impatient, surprised and in front of something they like and hate!
Good work and i can´t wait for Into the Nexus too!!
My pleasure and can´t wait for next R&C fanarts of yours! :3
And yeah, it really sounds interesting and promising and I´m curious about since the game is epilogue for the future series, this could be a last chapter of Ratchet´s guts to find his races.

subset: commoncrawl


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SIOUX FALLS — Winner forced Tri-Valley to its second straight double-overtime game Friday, allowing the Mustangs to get comfortable in extra minutes.
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Dictionary » B » B-lymphocytes B-lymphocytesB-lymphocytes lymphoid cells concerned with humoral immunity. They are short-lived cells resembling bursa-derived lymphocytes of birds in their production of immunoglobulin upon appropriate stimulation. Please contribute to this project, if you have more information about this term feel free to edit this page
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News came down late Thursday afternoon that Dwight Howard is finally leaving the Orlando Magic, heading to the Los Angeles Lakers in part of a four team deal involving the Magic, Lakers, Philadelphia 76ers and Denver Nuggets. While it's a major coupe for L.A., and Celtics fans are never happy when good things happen for the [...]
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Maines Paper & Food Service, Inc. To Hire 100 Employees For New Terrell Location
TERRELL, Texas, Oct. 20, 2011 /PRNewswire/ -- Maines Paper & Food Service, Inc., one of the leading independent foodservice distributors in the country, plans to hire more than 100 employees when in full operation, and plans to begin shipping operations at their new Terrell distribution center in April.
The official groundbreaking 9 a.m. on Friday, Oct. 21, 2011 at the Terrell Airport Business Park. Texas Agricultural Commissioner Todd Staples will be the guest speaker.
The distribution center is further evidence of Maines' commitment to excellence in customer service. The 100,000-square-foot facility will service 196 restaurants spanning across the states of Texas, Arkansas, Louisiana, Missouri and Oklahoma. This facility will be LEED-certified and their first distribution center west of the Mississippi River. At full build out, the facility will be in excess of 400,000 square feet and employ up to 400.
"Having a company like Maines expand their operations in Terrell's Airport Business Park is great news for our city, the Dallas/Fort Worth Metroplex and the State of Texas," states Mayor Hal Richards.
Terry Walsh, Maines' Executive Vice President and COO, said the company relocated one of its best managers to Terrell. " Jeff Warren, General Manager, will manage the construction and then oversee the operations starting in April 2012." " Terrell is the perfect place for our new center," explained Chris Mellon, President and CEO of Maines. "It is an ideal strategic distribution location. The Terrell area offers an impressive incentive package and a superb workforce." With over 100 positions to fill, including CDLA drivers, warehouse associates and customer service associates. All positions will be filled in March. Interested applicants can view the opportunities online at www.maines.net or contact Jeff Warren directly at the Terrell Chamber of Commerce. "We are excited to be in Texas as we continue our national expansion," said Chairman, Bill Maines. "We're impressed with the business-friendly leadership, led by Terrell Mayor Hal Richards. Terrell is a first-class town and we are thrilled to be a part of it." About Maines Paper & Food Service, Inc. Maines is one of the largest independent foodservice distributors in the country with a 90-year history of helping restaurant owners and managers maximize the performance. For more information log onto maines.net Stephanie Wyatt Maines Headquarters email@example.com 607 779.1200 SOURCE Maines Paper & Food Service, Inc.Check Out Our Best Services for Investors
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text: fav.me/d5gz6t8 I like to give away now times you need not give me back a llama...have fun! no problem and youre welcome I'm member of: #GetWatchers help artists to share their creativity, increase their audience and get more feedback by getting more exposure and pageviews. If you want more exposure of your arts, constructive critics, watchers and/or if you would like to discover new talented artists, come join us . |


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Maine |
Harbour Towne Inn On The Waterfront Boothbay Harbor, ME |
The Harbour Towne Inn is located in the heart of beautiful Boothbay Harbor, Maine - we have been welcoming guests with classic Down East hospitality for three generations. The newly refurbished waterfront bed and breakfast is within a historic Victorian home offering water views from the decks. Our quiet and convenient location is only a few steps from Downtown Boothbay Harbor's popular shops, restaurants and art galleries. Local activities include boating, whale watching, and scenic cruises. |


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Dictionary » H » Hip bones Hip bonesa large flat bone formed by the fusion of the ilium, ischium, and pubis (in the adult), constituting the lateral half of the pelvis; it articulates with its fellow anteriorly, with the sacrum posteriorly, and with the femur laterally. Synonym: os coxae, coxa, coxal bone, innominate bone, os innominatum. Please contribute to this project, if you have more information about this term feel free to edit this page Results from our forumTrying to prove height increase...need help 20140723_121159.jpg So I think I have proof of concept for my finger which are long bones but there are some differences between finger bones and long bones. I need you to validate that the finger on the right hand is longer. I have some before pictures but it's ...
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can anyone identify these bonesCan anyone identify these bones ? I found them In a graveyard beside a partially burried blanket. Just wondering if I should report it or if its an animal.
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Can anyone identify this bone...The only bones I can think of that curve like that are phalanges supporting claws or possibly part of a rib, although these ar not normally so smoothly round, Are you sure it is a bone?
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reproductive sytem... effects across the whole body. For example testosterone has been shown to prevent the onset of osteoporosis (the formation of air cavities in the bones). You can Google the effects of these two and find a huge amount of material - take your pick! Sex hormones are meant to have an ancient role ...
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Re: Need help identifying these bonesI need help identifying these bones. I discovered then off the coast of the Rappahannock in Whitestone VA.
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text: 000 WGUS82 KJAX 281356 FLSJAX FLOOD STATEMENT NATIONAL WEATHER SERVICE JACKSONVILLE FL 856 AM EST SAT FEB 28 2015 ...The flood warning continues for the following rivers in Georgia... Altamaha River At Baxley affecting Appling...Tattnall...Toombs and Wayne Counties PRECAUTIONARY/PREPAREDNESS ACTIONS... All persons with interest along the river should monitor the latest forecasts...And be prepared to take necessary precautions to protect life and property. Do not drive cars through flooded areas. If you see flood waters...Remember to turn around and do not drown. For graphical hydrologic information...Please go to weather.gov and click on your state. Select Rivers and Lakes under current weather. The Advanced Hydrologic Prediction Service page provides current and forecast river information...Gage locations...Impacts... and historical crest information for all forecast points by clicking on each point. && GAC001-267-279-305-011356- /O.EXT.KJAX.FL.W.0003.150301T0000Z-000000T0000Z/ /BAXG1.1.ER.150301T0000Z.150305T1200Z.000000T0000Z.NO/ 856 AM EST SAT FEB 28 2015 The Flood Warning continues for The Altamaha River At Baxley. * from this evening until further notice...Or until the warning is cancelled. * At 8:45 AM Saturday the stage was 74.2 feet. * Minor flooding is forecast. * Flood stage is 74.5 feet. * Forecast...Rise above flood stage by this evening and continue to rise to near 76.0 feet by Thursday morning. additional rises are possible thereafter. * Impact...At 74.5 feet...Minor flooding of timber and agricultural land can be expected. && FLD OBSERVED 7AM EST / 8AM EDT FORECAST LOCATION STG STG DAY TIME Sun Mon Tue Wed Thu ALTAMAHA Baxley 74 74.2 Sat 09 AM 74.8 75.3 75.5 75.7 76.0 && LAT...LON 3194 8243 3188 8222 3176 8199 3183 8195 3193 8213 3200 8243 $$


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About a year after we redid the landscape at our house, Florida native style, the landscaper we worked with sent Eric the classic book on environmental thinking for the young: Dr. Seuss's The Lorax. You remember it, I'm sure: truffula trees, brown bar-ba-loots, swomee-swans, all cavorting in a pre-development paradise.
Well, one of the things I've always sort of enjoyed about south Florida is how Seussian the landscape is. Even more than southern California with its joshua trees (although those are native), the flora here in Florida seems to have come straight from the pages of Dr. Seuss.


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Maggie Gyllenhaal Kicks Off Climate Matters Video Contest
Maggie Gyllenhaal, one of the stars in this summer's biggest blockbuster The Dark Knight, is teaming up with the organizations Brighter Planet and 1Sky to launch the "Climate Matters Video
The Green Quote: Natalie Portman Helps Makes The World A Better Place
"Look – the one thing that I find really shocking, I mean, that is something you don't learn in school that I was really surprised to find out about, was
Fast Food Ban In South LA Means Less Places For Lohan To Eat
Add hamburgers and french fries to the endangered list! Residents of South L.A. will have fewer fast food options in the coming year. The Los Angeles City Council has voted
Heather Mills To Make Appearance At The Animal Rights 2008 National Conference
Go get your calender…I'll wait. OK, got it? Great, now turn to August 14th-18th. What does it say? If you have scribbled down anything OTHER than "Attend Animal Rights 2008
JK Rowling's New "Beedle the Bard" Book To Benefit Charity
As expected, Harry Potter author JK Rowling is taking The Tales of Beedle the Bard into mass production. The collection of fairy tales, which was referenced in Deathly Hollows, was
Kanye West Does Think Of The Animals!
So I think we are all allowed at least one gratuitous post am I right? You've probably already seen the 'Be Kanye' ads or the official Be Kanye page that
If Ryan Cabrera Can Convert A Prius Hybrid, So Can You
Ryan Cabrera looks all hip and grown up. I remember him as a fresh faced kid hanging out with Ashley Simpson some years back – time flies. While promoting his


text:
Registration takes less than 1 minute.
Great Lakes vessel operators, shippers seek second large icebreaker for U.S. Coast Guard.
Justice Department said sentence is first against an executive in its roll-on/roll-off shipping investigation.
Union reportedly has comprehensive offer.
On-time performance is being hampered by congestion at West Coast ports.
DHS funding crisis means the agency will have to focus resources on border security, cargo clearance
The revised agreement would allow collaboration on congestion, transportation infrastructure needs and pollution mitigation.


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"Ten government forces died and 14 others were injured in the attack today. Four lawmakers were also injured. Seven of the fighters who attacked the house were also killed as you see their bodies," Kasim Ahmed Roble, a police spokesman, told reporters at the scene. Roble made no mention of any civilian casualties. A spokesman for al Shabaab, Sheikh Abdiasis Abu Musab, said that the group's fighters had killed 30 people. "We killed 30 from the AU (African Union) and from the various forces of the so-called Somali government," he said.
The al Shabaab estimate of the death toll was not independently verifiable. Reuters witnesses saw four bodies at the scene and saw a soldier fall from a rooftop after being shot. Reuters television pictures showed a large pool of blood near a blast site, and a man with his shirt drenched in blood running away from the scene. The fighting continued for hours after the initial explosion, with gunfire and smaller blasts being heard around the parliament.
"We are behind the suicide bombing, explosions and the fighting inside the so-called Somali parliament house, and still heavy fighting is going on inside," said the al Shabaab spokesman. The African Union Mission in Somalia (AMISOM) said in a statement that all the lawmakers who were in parliament before the attack were safely evacuated.
The attack on parliament, a building about 300 metres (yards) from the president's palace that is guarded by African Union peacekeepers and Somali forces, showed that the al Qaeda-linked group remains capable of hitting the centre of Mogadishu despite being pushed out of the capital two years ago. "The terrorists have once again shown that they are against all Somalis, by killing our innocent brothers and sisters. These cowardly, despicable actions are not a demonstration of the true Islamic faith," said Prime Minister Abdiweli Sheikh Ahmed.
Nicholas Kay, Special Representative of the UN Secretary-General for Somalia, said: "The Federal Parliament represents the people of Somalia and their hopes and aspirations for a peaceful and stable future. Today's attack is an attack against the people of Somalia for which there can be no justification." Somalia's government is struggling to impose any sense of order, more than two decades after the fall of dictator Mohamed Siad Barre tipped the country into chaos. In February at least 11 people were killed when al Shabaab attacked the presidential compound. In April they killed two lawmakers.
A Western diplomat who has worked with regional intelligence agencies said the attack would add to pressure on President Hassan Sheikh Mohamud from about 100 parliamentarians who last month called for him to be impeached over worsening security. "The federal government is exercising no control," the diplomat said. "Those ... in parliament will start asking questions: what is this guy achieving?" The diplomat said the attack showed that a surge by the African Union peacekeeping troops had not weakened al Shabaab's capacity to wage asymmetric warfare in the capital, where co-ordination between Somali and foreign intelligence agencies is poor. "Because intelligence is fragmented, al Shabaab is slipping through the net," said the diplomat. "They are becoming more dangerous.
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Celebrity fashion designer Rachel Zoe ventures into e-commerce
Zoe's new quarterly subscription service costs $100 per shipment and will feature at least one item sold at significantly below cost.
1 - 10 of 34 Results
The luxury retailer will launch tomorrow a user-generated section of its site called #SaksStyle that aggregates shoppers' selfies shared on Instagram, Tumblr, Facebook and Twitter. And the images will be 'shoppable.'
Posted 09/03/2014Zak StamborPost a comment
While rising steadily in its own retail sales, the women's fashion apparel brand designer and manufacturer carefully manages its wholesale sales through retailers, it says.
Posted 05/27/2014Paul DemeryPost a comment
Suppliers of retail store display equipment have new ways to connect with managers responsible for store displays through a new online business exchange launched today, A.R.E. Supplier Connect.
Posted 03/03/2014Paul DemeryPost a comment
Related Content:H&m | Online Merchandising Trends
Briggs joins Saks Fifth Avenue with a background in luxury branding, marketing and visual.
Posted 12/19/2013
Related Content:Saks Fifth Avenue
The retail chain already delivers orders and responds to inquiries quickly.
Posted 11/21/2013Mark BrohanPost a comment
Related Content:J.c.penney | Penney | Jcp
It resolves customer problems by e-mail better than other retailers.
Posted 09/18/2013Amy DustoPost a comment
Related Content:Stellaservice | Shopbop
The Canadian retail chain also names the future president of Saks Fifth Avenue.
Posted 09/17/2013Bill BriggsPost a comment
The retail chain had filed for an initial public offering in June.
Posted 09/09/2013Zak StamborPost a comment
Related Content:Neiman Marcus
Saks.com and the Saks Fifth Avenue department store chain will continue operations.
Posted 07/29/2013Allison EnrightPost a comment
Related Content:Saks Ecommerce | Saks Fifth Avenue | International Shipping
The merchant is encouraging in-store shoppers to use its new mobile app.
Posted 09/19/2012Bill SiwickiPost a comment
Related Content:Saks Fifth Avenue | Wifi


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27 year old woman | Batavia, NY, USA | Seeking men 25-40 within 50 miles
It's a good life.
I've lived the majority of my life in New York, except for brief breaks enjoy life as an ex-pat in Belgium and France. I love Europe and the endless adventures that it provides: hopping on trains to whatever destination, or renting a car and driving until you can't go any further. My family, my friends, drinking wine at the foot of the Eiffel Tower, walking along the Greystones, impromptu pub sing-a-longs, airplanes, white wine, photobooths, photography, waiting in airports, long train rides, new friends, new experiences, sushi, Mad Men, perfume, Red Velvet, baking, pretending to be a TV chef, and my St. Bernard: these are a few of my favourite things. I live for yoga. The best weekends I've ever had have been on a boat. I love craft beers and the Niagara Wine Trail. Outdoor concerts are the things that can make a summer. I dream of getting my pilot's license and learning to ride a motorcycle. Someone once described me as YOLOing before YOLO was a thing. That doesn't make me hate that term any less. I'm an up-for-anything, try-almost-anything-once type of person, hard-working, well-traveled, adventurous with a thirst to experience everything that the world has to offer. My ideal mate would understand the importance of family, the need for travel and new places, a desire to try new foods and an ability to be okay with just staying in bed on a rainy Sunday afternoon. Stability with a really good grip on reality and how things work in the world, ambitious, competent, compassionate. Someone who expects me to be more than what I already am and pushes me constantly to be better and doesn't humor me, or pretend. Someone sweet, spontaneous, courteous and who knows how to act like an adult when they're supposed to. A good sense of humour and intellect would be the icing on the cake.
Coffee and conversation , Cooking , Dining out , Gardening/Landscaping , Hobbies and crafts , Movies/Videos , Music and concerts , Performing arts , Political interests , Shopping/Antiques , Travel/Sightseeing , Volunteering , Watching sports , Wine tasting
Cycling, Running, Swimming, Yoga
I exercise 3-4 times per week.
I have Dogs
Brennan's, Colter Bay, Thirsty Buffalo, Frog Hair, Wasabi, The Blue Monk
All music from Johnny Cash to The Tragically Hip to Bell X1. I enjoy trashy television and thought-provoking movies. I love brewery tours and pub quiz nights. Also, netflix always recommends steamy foreign dramas for me, so take that as you will
I love art galleries, movies that do not have too much gore, boating when the weather is appropriate, walking through Letchworth, playing with my puppy and finding new ways to look at the world through the viewfinder on my camera.
Serena and I can't wait to see how they adapt it into a movie!
Her | What she is looking for | |
Appearance | ||
Height: | 5'10" (177cms) | 6'0" (182cms) to 8'11" (271cms) |
Body type: | Full-figured | No preference |
Eyes: | Blue | No preference |
Hair: | Dark brown | Black, Light brown, Dark brown |
Lifestyle | ||
Smoke: | No Way | No Way, Occasionally |
Drink: | Social Drinker | No preference |
Occupation: | Legal | No preference |
Income: | $35,001 to $50,000 | $35,001 to $50,000, $50,001 to $75,000, $75,001 to $100,000, $100,001 to $150,000, $150,001+ |
Relationship: | Never Married | No preference |
Have kids: | No | No, Yes, and they live away from home |
Wants kids: | Someday | Someday, Not sure |
Background | ||
Ethnicity: | White / Caucasian | White / Caucasian |
Faith: | Agnostic | No preference |
Languages: | English, French, German | English |
Education: | Bachelors degree | Bachelors degree, Graduate degree, PhD / Post Doctoral |


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When you're hot, you're hot! This classic is for all you classic Giants (and Eli Manning) fans - 'cause these dudes are smoldering! The future? This image should serve as both an icon and - when times are tough - reminder of how things can change.
This great custom product in heather gray from our department Men and T-Shirt category can be printed just for you in the size and style you like. The design Giants Helmet Smoldering was created by cappyjames and uploaded to Spreadshirt for you to have printed on a T-Shirt. Next to a host of other great customized designs, you can find this Giants Helmet Smoldering design in Spreadshirt's Football category. If you are interested in topics like 10 or Giants, then you'll love Giants Helmet Smoldering designs by cappyjames. Spreadshirt also offers a huge selection of custom t-shirts, hoodies and accessories for the category Football as well as top New clothing. cappyjames's designs have already impressed many of our customers, so we are sure that we've got right stuff for you! Spreadshirt is your destination for great designs of all kinds – whether it's 10, Giants or New. We are sure that you will find something nice in our shop, there are many amazing custom Football t-shirts, hoodies, or accessories for you to find! Tags: Design ID: 11743149
Retired Public Defender with a lifetime of bad jokes! Divorced father of four girls... i.e. have never been right about anything... Big heart, not too dumb, but forgets a lot.
Humor, politics (until everyone gets angry?), artsy stuff, clever stuff, stuff, and other things...
Nah Rosa Parks Quote T-Shirts Designed by The Shirt Yurt
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text:
Hey guys,
I finally decided to get a new bike and will be doing all kinds of riding - all mountain.
Question is:
[SIZE="4"]Specialized FSR XC Comp vs GT Force 3.0[/SIZE]
The GT Force 3.0 is $100 cheaper, but that doesn't matter.
Please help!!!
Thank you so much!
Results 1 to 2 of 2
2015 LIGHTS SHOOTOUTContestsHot Deals See All Hot Deals >>Latest Mountain Bike ArticlesMTBR on Facebook |


text:
Roddy White
Roddy White isn't happy about criticism of Aldon Smith
emerge. Falcons wide receiver Roddy White somewhat defended Smith, 23, who ..... have all driven drunk no doubt — Roddy White (@roddywhiteTV) September 22 ..... it you just got away with it — Roddy White (@roddywhiteTV) September 22
Pereira: Refs right to pick up flag on Mundy
A flag on a big hit on Roddy White was picked up, and Mike Pereira thinks the crew in Atlanta got it right.
Roddy White vows to 'Favre' Tony Gonzalez
Updated NOV 20, 2013 7:02a ET Football The Falcons star intends to bring the entire Atlanta Falcons team to California and bring Tony Gonzalez back for another season. Brett Favre \ Tony Gonzalez \ Roddy White
Roddy White apoligizes to New Orleans
Updated NOV 20, 2013 7:01a ET Football The Falcons wide receiver's mea culpa to The Big Easy. Roddy White
Falcons WR Roddy White arrested for outstanding traffic violation
Atlanta Falcons wide receiver Roddy White was arrested in the Atlanta area early Tuesday morning for an outstanding traffic violation. A Gwinnett (Ga.) County police
Panthers, Falcons head in opposite directions
make sure we do a good job of making sure that we take care of business at home.'' MIX 'EM UP: Falcons receiver Roddy White said the offense was surprised by the zone coverage Carolina deployed and was disappointed that Atlanta never seemed
Falcons hope to avoid another injury in secondary
RB Steven Jackson (quadriceps) and reserve CB Josh Wilson (hamstring) also didn't participate. ... WR Roddy White (ankle), WR Harry Douglas (foot) and RG Jon Asamoah (back) were limited in practice. --- AP NFL websites
Falcons QB Matt Ryan pushing to win for coach Mike Smith
memorable.'' Notes: Pro Bowl WR Julio Jones (hip/oblique) did not participate in practice. Also held out were WR Roddy White (ankle), RB Steven Jackson (quadriceps) and reserve CB Josh Wilson (hamstring). ... WR Harry Douglas (foot
Panthers-Falcons Capsule
straight opponents under 60 yards rushing. ... PK Matt Bryant leads NFL with seven FGs of 50 yards or longer. ... WR Roddy White has at least one TD in last five games against Panthers. --- AP NFL website: www.pro32.ap.org and http
Things are getting a little heated between Falcons and Saints
running high. We've got a little pushing and shoving here that ends with Jimmy Graham getting his head pushed back. Roddy White takes exception to what the Saints are doing and starts getting handsy. It's not really clear what Kenny Still is


text:
Winchester® USA Pistol .38 Special 125 Grain JFP 50 rounds
Item # WX2-95180
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You must be 21 years of age to purchase this item.
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Shipping restrictions apply to this product under some jurisdictions. Known age, state and local shipping requirements are applied at checkout and may result in changes to an order.
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Pistol and Revolver Ammunition
- Cartridge: .38 Special Ammo
- Bullet Wt. Grains: 125
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- Muzzle Energy (ft.-lbs.): 01
- Rounds: 50
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text:
What does WMVI mean in Professional Organizations?
This page is about the meanings of the acronym/abbreviation/shorthand WMVI in the Business field in general and in the Professional Organizations terminology in particular.
West Michigan Chapter of the Vibration Institute
Organizations --> Professional
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Discuss this WMVI abbreviation with the community:
Use the citation below to add this abbreviation to your bibliography:
"WMVI." Abbreviations.com. STANDS4 LLC, 2015. Web. 26 Feb. 2015. <http://www.abbreviations.com/term/243516>.
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text:
Featured news
Video: How the grid will be hacked
There is a never ending stream of vulnerabilities in the mechanical and cyber systems that support the grid.
Wireless hack attacks target critical infrastructure
Critical infrastructure control systems are at risk from wireless attacks carried out over Software Defined Radio (SDR), according to Digital Assurance.
Oil and gas infrastructure cybersecurity spending to increase
As a highly critical sector, the oil and gas infrastructure should be one of the most secure, both physically and digitally.
ENISA reports on smart grid cyber security measures
The EUs cyber security agency ENISA has investigated the challenges for baseline smart grids protection in Europe.
Smart grid ICS cyber security market to grow
Smart grid industrial control systems (ICS) remain in a state of flux.
(IN)SECURE Magazine issue 45 released
(IN)SECURE Magazine is a free digital security publication discussing some of the hottest information security topics. Learn about personal data bankruptcy and the cost of privacy, security and compliance, delivering digital security to a mobile world, and much more.
HITB Haxpo: Connecting hackers, makers, builders and breakers
Youri van der Zwart is running HITB Haxpo - a free exhibition for hackers, makers and software developers taking place alongside the Hack In The Box conference in Amsterdam, 27th - 29th May 2015.
Black hole routing: Not a silver bullet for DDoS protection
As ISPs, hosting providers and online enterprises around the world continue suffering the effects of DDoS attacks, often the discussions that follow are, What is the best way to defend our networks and our customers against an attack?
Compromised cPanel "Account Suspended" pages redirect to exploit kit
The code redirects visitors to another URL where the Fiesta exploit kit is hosted, which then tries to detect and exploit several vulnerabilities in various software. If it succeeds, the visitors are saddled with a banking Trojan.
Real MITM attacks enabled by Komodia's software might have already happened
EFF researchers showed that MITM attacks taking advantage of software using Komodia's encryption libraries have probably been executed in the wild.
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text: WWDC: Apple rumor checklist – updated by Philip Elmer-DeWitt @FortuneMagazine June 2, 2014, 9:23 AM EST E-mail Tweet Facebook Google Plus Linkedin Share icons Mark Gurman was 18 years old and still in high school when he scooped the Wall Street Journal, the New York Times and several dozen full-time bloggers with a series of eight predictions about what Apple AAPL was going to unveil at WWDC 2012. Seven of them were correct. We're still waiting on the eighth — Macbook Airs and iMacs with Retina displays. (See The high-school blogger who's been cracking Apple's secrets.) Gurman is back this year with a bumper crop of WWDC predictions — 36 by my count. Here's the checklist I made from his most recent rumor round-up. He edited it a few hours before the keynote, so he's got no excuses. We'll revisit the checklist after the event to see how well he did. UPDATE: Here's the annotated checklist:


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Don's Johns, Inc., established in 1964 by Thelma and Don Rainwater, was one of the first temporary restroom companies on the East Coast. We've been in business for more than 40 years, still family owned and operated, and we are the leading provider of portable restrooms, toilets, restroom trailers and sanitation services in the Virginia, Maryland, and Washington, DC, Metropolitan region. |


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One of the great Victorians poets, Tennyson's genius is expressed through the precision and delicacy of the language of his lyrical poems.
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I remember when I found out that I was pregnant with my youngest son, I stumbled upon the pregnancy forum on Baby Center. Baby center has a birth club for the birth month and year that my baby was due so that you could connect with mothers who were experiencing the same things that you were. It almost seems like yesterday that I was busy surfing through the forums and counting down the weeks until my baby was born. During those first few weeks, I saw countless posts about mother's who were frantic because they found out they were pregnant. Most of these women didn't realize that they were pregnant until they were half way through their first trimester and they wanted to know how harmful cigarette smoking could be for their baby. There is no right answer for this question because there are a number of various consequences for smoking during pregnancy.
Pregnant mothers who smoke cigarettes during their pregnancy increases your risks for potential problems. Also, cigarette smoking is linked to fertility issues. If you smoke during pregnancy you increase your risk for miscarrage, give birth to a premature baby, your membranes rupture prematurly, low birth weight, breathing problems at birth, allergies and ashtma, frequent infections, physical and mental deformaties, born stillborn, or death after delivery. Continue reading Smoking During Pregnancy – What Risks Are Involved For Me and My Baby? at Uplifting Families.
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Prepare for a meeting of the teen pop titans.
One of the many surprises set for Thursday night's 1999 MTV Video Music Awards broadcast was let out of the bag on "Total Request Live" when 'N Sync talked about teaming with Britney Spears for a performance on the awards show.
"We're doing 'Tearin' Up My Heart,' but it's not the usual 'Tearin' Up My Heart,'"
'N Sync's Lance told VJ Dave Holmes during Wednesday's "TRL."
"Basically, they were like, 'We want to do a pop set, blah, blah, blah,' and we were like, 'Sure, we'll do it'"
J.C. added. "But we wanted to do it a little bit different, and you'll see what happens tonight."
"There's a whole concept behind the performance, so we wanted to make sure it wasn't just, 'Hi, we're here,'"
Justin explained. "It's a whole concept, so it'll be good."
But precisely what that concept is remains to be seen. "We're not going to tell you,"
Justin said. "You'll just have to watch."
'N Sync, Britney Spears, and the rest of this year's VMA crop will start filing into New York's Metropolitan Opera House at 6:30 p.m. Thursday evening, which is when MTV News' coverage begins. You can tune into MTV News' pre-show coverage at 6:30, and check in with MTV News Online for updates throughout the night.
Catch all the sizzlin', star-packed VMA action direct from Miami on August 28. MTV News' preshow kicks things off at 6:30 p.m. ET/PT, followed by big show at 8 p.m.
Vote for the Viewer's Choice Award, see a timeline of MTV Video Music Awards history, and relive highlights of last year's event. And for more MTV Video Music Awards news, check out the MTV News VMA archive.


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2614 West Rio Vista Street
Almost all errands require a car.
Flat as a pancake, minimal bike lanes.
Score Details
2614 West Rio Vista Street has a Walk Score of 1 out of 100. This location is a Car-Dependent neighborhood so almost all errands require a car.
This location is in Wichita.
The closest school is West Elementary School.
The closest grocery store is Johnson's General Stores.
Nearby restaurants include SUBWAY® Restaurants (Wal-Mart) and McDonald's.
2614 West Rio Vista Street is near Woodland Park, Oak Park and McAdams Park.
There are minimal bike lanes and the terrain is flat as a pancake. 2614 West Rio Vista Street is somewhat bikeable, there is minimal bike infrastructure.
Explore how far you can travel by car, bus, bike and foot from 2614 West Rio Vista Street.
View all Wichita apartments on a map.
Popular apartment searches include fireplace, hardwood floor and pool. View all Wichita apartments and rentals.
This location is in the city of Wichita, KS. Wichita has an average Walk Score of 32 and has 382,368 residents.


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Australia–Solomon Islands relations
From Wikipedia, the free encyclopedia
Australia |
Solomon Islands |
Australia–Solomon Islands relations refers to foreign relations between Australia and Solomon Islands. Australia has a high commission in Honiara and Solomon Islands has a high commission in Canberra.
Australian aid[edit]
Australia is a significant foreign aid donor to the Solomon Islands. The Australian Government leads other nations as part of the Regional Assistance Mission to Solomon Islands. Over 7,000 Australian Defence Force members have been deployed in Solomons as part of this mission. [1]
In 2013, the Australian government committed $500 million to assisting Solomon Islands through the regional assistance mission in the following 4 years. [2]
This article about foreign relations is a stub. You can help Wikipedia by expanding it. |


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Academia Venezolana de la Lengua
From Wikipedia, the free encyclopedia
Abbreviation | AVL |
Formation | July 26, 1883 |
Type | NGO |
Purpose | Language Academy for Venezuelan Spanish |
Headquarters | Caracas |
Region served
Venezuela |
Association of Spanish Language Academies |
Website | avelengua.org.ve |
The Academia Venezolana de la Lengua (Spanish for Venezuelan Academy of Language) is an association of academics and experts on Venezuelan Spanish, the variant of the Spanish language in Venezuela. It was founded in Caracas on July 26, 1883. It is a member of the Association of Spanish Language Academies.
See also[edit]
- Category:Members of the Venezuelan Academy of Language (includes past members)
External links[edit]
This article about a linguistics organization is a stub. You can help Wikipedia by expanding it. |
This article about an organisation in Venezuela is a stub. You can help Wikipedia by expanding it. |


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Jon Stewart was looking forward to the end of congressional gridlock now that Republicans control both houses.
John Oliver, who once spurred his followers to crash the FCC's website demanding net neutrality, is positively gleeful over the news that the agency has approved rules to regulate the internet like a public utility.
Remember that time Joe Biden got close to Secretary of Defense Ashton Carter's wife?
Thanks to some mischievous editing from Jimmy Kimmel, these originally innocent TV clips will put your mind straight in the gutter.
White House
WASHINGTON (AP) — Amid appeals for bipartisanship, President Barack Obama in just three days has provoked Republicans on issues as disparate as immigration, Wall Street and the Keystone XL pipeline — a combative mix of defense and offense that underscores Washington's political realignment.
Defying the Republican-run Congress, President Barack Obama rejected a bill Tuesday to approve construction of the Keystone XL oil pipeline, wielding his veto power for only the third time in his presidency.
Joe Biden—everyone's favorite creeper and truth-bomb-dropper.
The president's interview with Re/code over the weekend touched on privacy issues, with Obama insisting with "almost complete confidence" that there have been no abuses of the government's vast surveillance program.
The Hill reports that Democrats are beginning to worry about Elizabeth Warren's gravitational pull on the party's direction. It's a black hole of progressivism, baby.
Rand Paul's speech at CPAC Friday felt a lot like a campaign rally—and the crowd left little room to doubt that they wanted it that way, breaking out into chants of "President Paul! President Paul!" at least three times over the course of his remarks.
OMAHA, Neb. (AP) — A federal judge blocked Nebraska's gay marriage ban on Monday, but the decision will not take effect for a week and the attorney general's office immediately appealed to the 8th U.S. Circuit Court of Appeals.
The New York Times reports that Leonard Nimoy, who was and always will be "Spock" of "Star Trek" fame, passed away at his California home Friday morning.
DEA agents seem to be scraping the bottom of the barrel when it comes to anti-pot arguments these days.
The Republican Party may be in for some serious generational shifts when it comes to drug policy.
Colorado's newly-released report on the first year of its legal marijuana industry describes a flourishing industry: recreational consumers purchased over 17 tons of marijuana buds, and medical marijuana patients nearly 50 tons.


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Health restored? Uncertainty in forecasting Thailand's economic outlook
In Laos, the government has reportedly already healed the economy from the economic flu.
In Laos, the government has reportedly already healed the economy from the economic flu.
The investment climate is the fundamental socio-economic framework in which firms operate – the macroeconomic and trade policies they face, the labor and financial markets in which they recruit and raise money, the available infrast
Another example of China's respectable growth, despite the global economic crisis, is apparent in this month's Fortune magazine, with its Global 500 list of the world's largest companies.
About a month ago, I came across Changemakers.com (via Change.org's Social Entrepreneurship blog), a neat website for people to connect and collaborate with others working – on all levels – to solve social problems. The website is an initiative of Ashoka, a nonprofit organization that works to support social entrepreneurship. Changemakers seems to act as a social network of sorts – through competitions, discussion forums and storytelling – for people who want to make a difference. Two aspects of the site quickly appealed to me.
Pongtip Puvacharoen works at the World Bank's East Asia and Pacific stand on the first day of Carbon Expo in Barcelona. |
Carbon finance sounds boring and technical and not much fun. However, it actually does a lot of good and can help fund critical environmental preservation projects as well as introduce clean and renewable technologies in both developed and developing countries.
It takes me just a few minutes to get to my office roughly two kilometers away. Before the Skytrain came along, the very same journey could take anywhere between 15-45 minutes. |
In terms of big newsworthy events in Asia, one of the biggest has to be the anti-government protests in Thailand. A relatively small number of protesters dramatically caused the cancellation of an ASEAN+3 meeting held in Pattaya this past weekend where 10 regional heads of state were evacuated. The World Bank President, as well as the head of the IMF and UN, were turned around at the airport in Bangkok. Although the protests around the country have effectively ended after martial law was declared and two protesters died, the damage of this may be longer-lasting. Although a discussion of the politics would be interesting, let's concentrate on the finance-related issues.


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More than three quarters of the small and disadvantaged businesses questioned in a recent study doubted the value of a Small Business Administration business development plan aimed at helping them garner federal contracts.
According to a study released by Washington, D.C.-based Eddy Communications Corp. in late January, 87 percent of the participants of the SBA's popular 8(a) business development program that aims to help small socially and economically disadvantaged businesses were not sure their participation had brought any advantages, even after completing nine-year terms in the program.
The study's author said it showed that the 8(a) program didn't adequately prepare new contractors for their growth, from a small business into the larger, more competitive environment for federal contracts.
"We knew about the challenges of 8 (a) graduates face when they move into full and open competition—for years, many of our clients have sought us out to help them navigate that growth stage," said Jennifer Eddy, founder and chief strategist of Eddy Communications. "But the results of this study reveal how overwhelming the government market can be to newcomers who don't have anyone in their corner."
The 8(a) set-aside is one of several SBA programs aimed at helping socially and economically disadvantaged small businesses get federal contracts.
The program is divided into two phases -- a four-year developmental stage and a five-year transition stage -- in which small businesses are certified and participate.
Less than half of survey participants said they would confidently recommend the program to a friend in their industry, and nearly 10 percent said they self-terminated their affiliation with the 8(a) program early after determining the burdens of participation outweighed the benefits.
For its part, the SBA said in its fiscal 2014 budget justification that from January 2009 to January 2013, it has supported more than $106 billion in lending to more than 193,000 small businesses. from January 2009 to September 2012, the agency said it helped counsel and train more than three million small business owners and entrepreneurs through its resource partner network and through September 2011 helped small businesses access more than $286.3 billion in federal contracts. That is, SBA said, $32 billion more in small business contracting than the previous three years, even as overall contract spending decreased.
About the Author
Mark Rockwell is a staff writer covering acquisition, procurement and homeland security. Contact him at email@example.com or follow him on Twitter at @MRockwell4.
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President Obama's proposal to boost government coordination with the private sector got a warm welcome in the House Homeland Security Committee.
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Meet the women and men who are doing great things in federal IT.
The law's certification and approval provisions empower CIOs to end outdated software development projects, says Agilex's Roger Baker.
The new chairman of the House Oversight and Government Reform Committee extends an olive branch to the minority, but keeps subpoena power for himself.
FCW investigated efforts by the departments of Defense and Veterans Affairs to improve a joint data repository on military and veteran suicides. Something as impersonal and mundane as incomplete datasets could be exacerbating a national tragedy.
Despite delays, the program is at a critical point for determining the ultimate impact of cloud technology in the government space.
In an interview with FCW, the Department of Veterans Affairs' chief technology officer talks about overhauling the digital experience for VA customers.
The National Information Exchange Model's usefulness extends far beyond its origins in justice and law enforcement.
Alaskan. Native Companies that are in the 8a program further muddy the water with the ridiculous advantages they have, like spinning off another 8a after they graduate. Great example - look up CSC's current protégés, and the first one listed is Arctic Slope Management Services, an 8a spinoff off of the parent company ASRC, a BILLION dollar company. If they have a parent organization that is bringing over a billion dollars, why do they need to be in a Mentor Protege with CSC???? I hope this comment gets posted so more people see this.
Unfortunately, the SBA has abandoned small business, and is now more unhelpful than anything. 28 million of the 28.2 million businesses in America, 98%, have 1-19 employees. They need loans of $50-$200,000. In 2007, the average SBA loan was $180,000 and 24% of all SBA loans were under $100,000. Today, the average SBA loan is $485,000 and only 9% are under $100k, and declining. The 98% have been left behind while the SBA supports the 2%. Almost every time the SBA gives out a loan now, it goes to giant businesses who use it to crush the small ones who can't squeeze a dime from the SBA's banks. How did we get here? In the last five years under Karen Mills, the SBA aggressively ad proactively expanded the definition of "small" to allow tens of thousands of very large businesses (500+ employees, $30+ million in revenue) to be reclassified as small. As a result, the banks and the SBA can now give a lot fewer loans to much bigger businesses, and still blow their horn that they support small businesses. Nothing could be farther from the truth. The banks and the venture capitalists (like Startup America) have been lobbying for years to get the SBA to abandon small business in favor of the 2%; the largest businesses in America. It's working. Their next step is to convince the politicians to fold the SBA into the Commerce Department, where it will get lost in the Commerce Department's focus on the top 1/2 of 1% of the largest businesses. The new SBA leader is a venture capitalist, so this is not likely to get fixed. The SBA world is upside down.
Big surprise. SBA is one of the gatekeepers and road blocks to legitimate small businesses. They make it almost impossible for any real business to qualify, expect you to line the pockets of the ex-staffer-now-consultants to make the list, and then don't do anything. Really too bad given they could be so helpful it they would, well, actually help....
get 8a certificate its waste time and money for the last two years i don't see no help


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Environmental assessment gets OK from Afton City Council
The book has finally closed on the paper maze that is the Environmental Assessment Worksheet for the Afton Center project.
The Afton City Council approved a negative declaration, or stated it adequate, at its Feb. 17 meeting.
The Afton Center EAW is a detailed assessment of all of the effects the proposed Afton Center project, a GJ&M proposed 48 luxury condo units and 12 retail spaces over three city blocks in downtown Afton.


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Salad Fingers A strange cartoon about a creature who loves to touch rusty metal.
Author Comments
My 4th Newgrounds release!! And the longest flash I've done yet (still not THAT long) Damn that was hard work! Anyways here it is, a sequel due to popular demand! I hope this will get as good a response as the first one. Please enjoy it.
Rated 5 / 5 stars
I have become a huge fan of Salad Fingers and this episode is currently my favorite. The style, ambience and vibe it gives off is genius; the perfect blend of ambient sounds, music, graphics and voice acting all perfectly mold the sense of fear. And of course this is not for everyone....it's just so irritating, but funny to read the reivews of 13 year olds who say David Firth needs help. Keep the episodes coming.
Rated 5 / 5 stars
dude i fucking love your shit. the Fat-Pie site doesnt work though :(. You should come out with murchandise for Salad Fingers it would be a huge hit. The first and 3rd one killed too!!!


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> The plot of Melmoth is confusing partly because we do not
I suppose that my immediate reaction of "spoiler alert!" makes me unworthy.
> learn the premise of the story until the end, where we
> discover that Melmoth had purchased extended life by
> selling his soul to the devil.
> Maturin, like C. S. Lewis, was an Irish Protestant, indeed
Then they weren't merely "Irish Protestants," the vast bulk of whom are
> a clergyman in the Church of Ireland.
nonconformists like Presbyterians. Maturin and Lewis were, then, both
adherents of the Established Church of Ireland, a very small body and a much
closer connection. Nicholson goes on to describe Wilde, Yeats, and Stephens
as "Irish Protestants" also, but he doesn't say whether they were Church of
Ireland or nonconfirmists. Insofar as sectarian distinctions are of any
importance, this is.
> It is not an accident that Wilde took the name of 'Melmoth'
And given the close association of the name of Melmoth with Wilde by Lewis's
> when he went into exile in France after imprisonment.
time, some comments on Lewis's opinion of Wilde - and could he separate the
man from the work? - might be relevant here, if they're available.
> I have not found explicit reference to Maturin in Lewis, but I
Each sentence here is a separate and distinct plunge into the Sea of Rampant
> believe it is a near certainty that he had read Maturin. Lewis
> read everything. He was especially interested in the fantastic,
> an area he read very widely in. The work of Irish Protestants
> held a deep interest for him, being an Irish Protestant himself
Speculation (or, if you prefer, a jump to the Isle of Conclusions).
> But what really connects Melmoth and Perelandra is the situation
Well, that settles it, then. Lewis couldn't possibly have come up with this
> they focus on. In both cases, we are presented with an isolated
> tropical island, where a beautiful but vulnerable young woman is
> besieged by a demonic male who wants to take from her something
> vital—her soul (it is not, as one might expect, sex that draws Melmoth
> and Weston to the woman in question).
on his own. (But in that case, where did Maturin get it?) I agree with
Jason: "he might have simply written a comparative piece and then wondered,
in his conclusion, whether Lewis might have read Melmoth."


text: Kid Angel - Rain Drops Feat. Otter [Prod. December] By Kid Angel Posted May 14, 2012 at 12:00 AM 0 0 0Comments 42 Views Share Tweet gplus thumblr pinterest 1 Rain Drops [Prod. December] Kid Angel feat. Otter Follow Artist Embed Me To Playlist Download Songs Mixtapes Videos 0 0 0 42 Powered By HOTNEWHIPHOPhotnewhiphop.com Edit Colors Full Player Simple Player Embed Script Copy Short Link Copy Iframe Copy Preview Full Player Simple Player Embed Code Copy NOTE: Minimum player width is 370 pixels, height is 50 pixels. Back Full Player Simple Player NOTE: Player width is 450 pixels, height is 50 pixels. Title Artist Back 01:23 04:56 Embed Code Copy


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A friend once told me (or was I just listening to people talk on a podcast? It doesn't matter) that getting into and staying in the habit of jogging is easy, as long as you never stop. While my sedentary husk knows almost nothing about exercise, I can testify it feels similar with drawing. If you haven't done it in a while, or if you haven't used a certain medium in some time, it's a major struggle to get yourself back up to speed. As with painting (or in this case digital painting), thinking in color is much different than the usual black and white ink drawing. Being someone who doesn't like cheap Photoshop tricks and shortcuts either, I'm stubbornly doing as much as I can manually, AKA the hard way. (If I could've built the world using colored pencils I would.) Historically I've gotten no shortage of nasty looks when I tell my peers and padawans about my distaste for digital trickery. So uncivilized! But enough of that. Here are some more placeholder backgrounds for Complex, our real time strategy farming, racing and casino mini-game collection. (One or more of those may not be accurate. Depends on our mood.) Today's first sketch, a somewhat chunky diorama, is the main character's home. This "room" is designed to take up about 4 screen widths and, as you can see, features world navigation where you can enter into the foreground and background to reach new areas. While it's a strictly 2D game, one of my personal goals is to make the world entirely comprehensible in a 3D space. So, one road leads to that barn in the background, which you can go into. You can enter the house or go a number of other places on the property through this "crossroad". I like that. This next image set takes up about two screen widths and is being carefully measured to create a "parallax" effect. We've still got tweaking to do, but basically that means that as you navigate from one side to the other, the far background will move at different speeds to (hopefully) strengthen the illusion of depth. The mountains might move a little bit, and the clouds even less. Not to sound like a broken record but it's super, super important to me that the audience feels like this is a real place and not just a playground of shapes. I will also say that those far mountains in the background contain something very important to the story. Complex, one might say. It all looks very rough, still, but I hope the "essence" of these locations translate well enough. Eventually I'm going to need to buy a stronger computer to let me "paint" the final versions at a larger size. We might even include animation (like grass swaying) if that's feasible. Time is going to be a factor here. Also, while I'm updating, here's a snapshot of some almost-finished Partial Quest art: That's it for this
afternoon. Time for a haircut and a shower. -_- February 8, 2015 - Horror Games Brainstorming Conference This month we plunge into
one of the darkest depths of games development, where light and hope
cannot reach:
The conference room. Can you survive the terror within on this
beautiful Sunday morning?! February 6, 2015 - If You're Into This Sort of Thing Here's another project I'm pleased to be working on. It's a mobile RPG (with hearty rogue-like dungeon-crawlish flavors) for release on android/iOS and all that. (I understand a PC port is also likely.) I'm handling art duties primarily, with some extra design on the side including story and minor gameplay/interface stuff. It's a sharp, sweet game which I've played, thoroughly enjoyed, and can easily recommend with a clear conscience. Stylistically, Partial Quest is going to look like an elaborately illustrated tabletop game. Almost every asset will have been drawn with pen on paper to give it a warm, hand-made feel. We're aiming for a release in March. In the meantime my teammate John has written up a project update at IndieDB and there should be more soon. If you like, please check it out! Tell your friends, even? I don't know, man. "This is a lot to be working on, isn't it?" One may ask. Perhaps, perhaps - but you'd be amazed at how much stamina you have when you're consumed by the creative spirit. These past few months have been the first in years when I've really stretched my creative muscles. With nothing but my own strength holding me back I feel like a wild artist monk, submerged in self-imposed exile with fire in his eyes, drawing up a storm. This is gonna be fun! =D February 4, 2015 - Happy Little Clouds Painting would be so much fun, if it weren't so scary. XD For anyone interested, below are two pretty little quasi-backgrounds which players of Complex will be able to explore someday. They're unfinished, obviously - the hope here is to capture the "essence" of the settings; concept art that you can walk around in. Tests have been promising. I look forward to the day when everything's ironed out to the point where I feel completely immersed in the game world. See that house in the background of the first image? You can go there. See that outhouse in the second image? You can go there too (if you catch my meaning). Maybe the game is actually a rocket ship simulator? I won't say. There should be more news
later this week about another project I'm part of. Something a bit more...
concise, but worthy all the same. I'm looking forward to this year! It's
been super busy so far and there's a lot of work to be done - it'll be a
delight to see some of this stuff bear fruit. (Note: I haven't forgotten
about Bite-Size... just... gotta... find some time. >_<) February 2 - 2015 - Sigh Hearing about bad news
happening to friends, I figured I should pull the link to the next journal
entry re: my trip to Japan last year. I'll put it back another time. =( |