text:
|Publication number
|Publication type
|Application number
|Publication date
|Oct 1, 2002
|Filing date
|Sep 9, 1998
|Priority date
|Sep 9, 1998
|Fee status
|Publication number
|Laurakay Bruhn, Michael P. Caren, Carol T. Schembri, Arthur Schleifer
|Original Assignee
|Agilent Technologies, Inc.
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (12), Referenced by (34), Classifications (34), Legal Events (8)
|External Links: USPTO, USPTO Assignment, Espacenet
The field of this invention is nucleic acid arrays.
"Biochips" or arrays of binding agents, such as oligonucleotides and peptides, have become an increasingly important tool in the biotechnology industry and related fields. These binding agent arrays, in which a plurality of binding agents are deposited onto a solid support surface in the form of an array or pattern, find use in a variety of applications, including gene expression analysis, drug screening, nucleic acid sequencing, mutation analysis, and the like.
Such arrays may be prepared in a number of different ways. For example, DNA arrays may be prepared manually by spotting DNA onto the surface of a substrate with a micro pipette. See Khrapko et al., DNA Sequence (1991) 1:375-388. Alternatively, the dot-blot approach, as well as the derivative slot-blot approach, may be employed in which a vacuum manifold transfers aqueous DNA samples from a plurality of wells to a substrate surface. In yet another method of producing arrays of biopolymeric molecules, a pin is dipped into a fluid sample of the biopolymeric compound and then contacted with the substrate surface. By using a plurality or array of pins, one can transfer a plurality of samples to the substrate surface at the same time. Alternatively, an array of capillaries can be used to produce biopolymeric arrays. See WO 95/35505. In another method of producing biopolymeric arrays, arrays of biopolymeric agents are "grown" on the surface of a substrate in discreet regions. See e.g. U.S. Pat. No. 5,143,854 and Fodor et al., Science (1991) 251:767-773. In yet another method of producing nucleic acid arrays, piezo inkjets are used to deposit nucleic acids on the surface of a substrate. See U.S. Pat. No. 5,658,802. However, the U.S. Pat. No. 5,658,802 patent states that with regard to thermal inkjet devices, "[t ]hermal ink jets, however, are very stressful on the dispensed fluid and the fluid must not be too aggressive to the heater element. Because of these constraints, thermal ink jets are generally unsuitable for dispensing applications other than those where the composition of the ink can be fully controlled." Col. 2, lines 14 to 19. Each of the above methods of producing arrays has certain disadvantages, e.g. complexity, non-reproducibility, etc.
Accordingly, there is continued interest in the development of new methods and devices for producing biopolymeric arrays. Of particular interest would be the development of a method for rapidly producing a nucleic acid array in which a minimal amount of the nucleic acid sample is used to produce reproducible nucleic acid spots.
Patents and patent applications describing arrays of biopolymeric compounds and methods for their fabrication include: U.S. Pat. Nos. 5,242,974; 5,384,261; 5,405,783; 5,412,087; 5,424,186; 5,429,807; 5,436,327; 5,445,934; 5,472,672; 5,527,681; 5,529,756; 5,545,531; 5,554,501; 5,556,752; 5,561,071; 5,599,695; 5,624,711; 5,639,603; 5,658,734; WO 93/17126; WO 95/11995; WO 95/35505; EP 742 287; and EP 799 897.
Other references of interest include: Lockhart et al., Nature Biotechnology (1996) 14: 1675-1680; Schena et al., Science (1995) 270: 467-470; Schena et al., Proc. Nat'l Acad. Sci. USA (1996)93:10614-10619; Shalon et al., Genome Res. (1996) 6: 639-645; Milosavljevic et al., Genome Res. (1996) 6:132-141; Nguyen et al., Genomics (1995)29: 207-216; Pi�tu et al., Genome Res. (1996) 6: 492-503; Zhao et al., Gene (1995) 166:207-213; Chalifour et al., Anal. Biochem. (1994) 216:299-304; Heller et al., Proc. Nat'l Acad. Sci. USA (1997) 94: 2150-2155; Khrapko et al., DNA Sequence (1991) 1:375-388; Lehrach et al., Hybridization Fingerprinting in Genome Mapping and Sequencing, Genome Analysis, Vol. 1 (Davies & Tilgham, eds)(Cold Spring Harbor Press) (1990) pp 39-81; Schema, M., BioAssays (1996) 18: 427-431; DeRisi et al., Nat. Genet. (1996) 14457-460; DeRisi et al., Science (1997) 278: 680-686; and Chen et al., Biomedical Optics (1997) 2: 364-374.
U.S. Patents disclosing the use of inkjet devices to dispense bio/chemical agents such as proteins and nucleic acids include: U.S. Pat. Nos. 4,877,745; 5,338,688; 5,474,796; 5,449,754; 5,658,802; and 5,700,637.
Methods and devices for depositing nucleic acids on a substrate surface are provided. In the subject methods, a thermal inkjet head loaded with fluid composition of the nucleic acid is positioned in opposing relationship to, e.g. over, a substrate surface. The thermal inkjet head is then actuated in a manner sufficient to expel a volume of the aqueous nucleic acid composition onto the substrate surface. The subject methods and devices find use in a variety of applications, particularly in the preparation of nucleic acid arrays, including arrays of oligonucleotides and polynucleotides, e.g. cDNA arrays.
FIGS. 1A and 1B provide representations of polyacrylamide gels of nucleic acids before and after firing from a thermal inkjet head.
FIGS. 2A and 2B provide a graphical representation of the hybridization characteristics of both inkjet fired and hand pipetted nucleic acids.
The term "nucleic acid" as used herein means a polymer composed of nucleotides, e.g. deoxyribonucleotides or ribonucleotides.
The terms "ribonucleic acid" and "RNA" as used herein means a polymer composed of ribonucleotides.
The terms "deoxyribonucleic acid" and "DNA" as used herein means a polymer composed of deoxyribonucleotides.
The term "oligonucleotide" as used herein denotes single stranded nucleotide multimers of from about 10 to up to about 100 nucleotides in length.
The term "polynucleotide" as used herein refers to single or double stranded polymer composed of nucleotide monomers of generally greater than 100 nucleotides in length and up to about 8,000 or more nucleotides in length.
The term "cDNA" as used herein means a complementary DNA molecule made as a copy of mRNA amplified using PCR for deposition on arrays. cDNAs can range from about 100 bp to about 8,000 bp, where average cDNAs are typically 1 to 2 kb in length.
Methods and devices for depositing a nucleic acid on the surface of a substrate are provided. In the subject methods, a thermal inkjet head loaded with a fluid composition of the nucleic acid is positioned in opposing relationship to, e.g. over, a substrate surface. The temperature of the heating element of the inkjet head is then raised such that a bubble is formed at the surface of the heating element and a volume of the nucleic acid fluid composition is expelled from the head onto the substrate surface. The subject methods and devices are particularly suited for use in the production of nucleic acid arrays, including oligonucleotide and polynucleotide arrays, e.g. cDNA arrays.
Before the subject invention is described further, it is to be understood that the invention is not limited to the particular embodiments of the invention described below, as variations of the particular embodiments may be made and still fall within the scope of the appended claims. It is also to be understood that the terminology employed is for the purpose of describing particular embodiments, and is not intended to be limiting. Instead, the scope of the present invention will be established by the appended claims.
In this specification and the appended claims, the singular forms "a," "an," and "the" include plural reference unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this invention belongs.
The subject invention is a method of depositing a nucleic acid on the surface of a substrate, as well as devices for practicing the method and the arrays produced thereby. Critical to the subject method is the use of a thermal inkjet head to deposit the nucleic acid on the substrate surface.
Thermal inkjet heads are well known in the art of conventional printing and document production. As is known to those of skill in the art, thermal inkj et heads typically have at least the following components: (a) an orifice; (b) a firing chamber; and (c) a heating element. Thermal inkjet heads and methods for their manufacture and use are described in a number of different U.S. Patents, including: U.S. Pat. Nos. 5,772,829; 5,745,128; 5,736,998; 5,736,995; 5,726,690; 5,714,989; 5,682,188; 5,677,577; 5,642,142; 5,636,441; 5,635,968; 5,635,966; 5,595,785; 5,477,255; 5,434,606; 5,426,458; 5,350,616; 5,341,160; 5,300,958; 5,229,785; 5,187,500; 5,167,776; 5,159,353; 5,122,812; and 4,791,435; the disclosures of which are herein incorporated by reference. Thermal inkjet heads finding use in the subject methods will generally have the following characteristics. The size of the orifice is sufficient to produce a spot of suitable dimensions on the substrate surface, where the orifice generally has a diameter (or exit diagonal depending on the specific format of the ink jet head) ranging from about 1 to 1000 μm, usually from about 5 to 100 μm and more usually from about 10 to 60 μm. The firing chamber has a volume ranging from about 1 pl to 10 nl, usually from about 10 pl to 5 nl and more usually from about 50 pl to 1.5 nl. The heating element will preferably be made out of a material that can deliver a quick energy pulse, where suitable materials include: TaAl and the like. The thermal element is capable of achieving temperatures sufficient to vaporize a sufficient volume of the nucleic acid composition in the firing chamber to produce a bubble of suitable dimensions upon actuation of the head. Generally, the heating element is capable of attaining temperatures of at least about 100� C., usually at least about 400� C. and more usually at least about 700� C., where the temperature achievable by the heating element may be as high as 1000� C. or higher. The device may also have a reservoir. When present, the reservoir will typically have a volume ranging from about 1 pl to 1 l, usually from about 0.5 μl to 10 μl and more usually from about 1 μto 5 μl. A variety of thermal inkjet heads are available commercially, where such devices include: the HP92261 A thermal inkjet head (available from Hewlett-Packard Co., Palo Alto Calif.), the HP 51645 A thermal inkjet head (available from Hewlett-Packard Co. Palo Alto Calif.), the inkjet produced by (Cannon Kabushiki Kaisha,Tokyo, Japan) and the like. Specific inkjet heads of interest are disclosed in U.S. Pat. Nos. 5,736,998 and 4,668,052, the disclosures of which are herein incorporated by reference.
In practicing the subject methods, the thermal inkjet device is loaded with a fluid nucleic acid composition or fluid. By fluid nucleic acid composition is meant a composition of nucleic acid molecules in combination with a carrier liquid, where the nucleic acid molecules will generally, though not necessarily, be dissolved in the carrier liquid such that the carrier liquid serves as a solvent. The carrier liquid may be one or more different fluids, including polar liquids, e.g. water, DMSO, formamide, and the like. In many instances, the carrier liquid will be aqueous, being either water alone or water in combination with a co-solvent. The nucleic acid fluid composition may also contain one or more additional agents, including: buffering agents, salts, metal cations, surfactants, enzymes, etc., where such agents may be present in the composition as a result of the way tin which the composition is prepared. For example, where the nucleic acid composition is prepared via preparative PCR techniques, the fluid composition that is loaded into the inkjet head may contain, in addition to water and the PCR produced product DNA, buffers, dNTPs, polymerase, salts, left over primers, and the like. The nucleic acid fluid compositions may be prepared in a number of different ways. Thus, the nucleic acid may be synthesized using phosphoramidite synthesis technology, preparative PCR, purification from naturally occurring sources, and the like, where such methods are well known to those of skill in the art.
The nucleic acid of the aqueous nucleic acid composition may vary considerably. As such, the nucleic acid may be DNA, RNA or some other synthetic type of nucleic acid in which the conventional backbone has been replaced with a non-naturally occurring or synthetic backbone, e.g. PNA and the like, and/or one or more of the conventional bases has been replaced with a synthetic base capable of participating in Watson-Crick type hydrogen bonding interactions. The length of the nucleic acid may vary and will be chosen depending on the article to be prepared using the subject method, e.g. the nature of the nucleic acid array to be produced. Thus, the nucleic acid may be an oligonucleotide or a polynucleotide. Of particular interest is the use of the subject methods to deposit oligonucleotides on the surface of a substrate, i.e. to produce oligonucleotide arrays. Also of particular interest is the use of the subject methods to deposit polynucleotides on the surface of a substrate. As indicated in the definitions section above, a polynucleotide is a polymer of nucleotides that is at least about 100 nt in length, usually at least about 120 nt in length and more usually at least about 150 nt in length, where the polynucleotide may be a long as 8,000 nt in length or longer, but in many embodiments will not exceed about 3,000 nt in length. The nucleic acid in many embodiments of particular interest is cDNA.
The nucleic acid composition or fluid may be loaded into the firing chamber and fluid reservoir using any convenient means. Thus, conventional methods of introducing ink into thermal inkjet heads may be employed. Where such methods are employed, following loading of the nucleic acid fluid into the inkjet head, it is often desirable to "prime" the device prior to use. One means of priming the device is to apply sufficient pressure to the fluid in the reservoir (or conversely negative pressure to the orifice) such that a volume of fluid is forced out of the orifice. Such priming methods are currently employed in the printing industry and thus are well known to those of skill in the art.
Alternatively, where minimal waste of the nucleic acid fluid is desired, e.g. where the fluid is an expensive or rare cDNA sample, the following method of loading the nucleic acid composition into the firing chamber and reservoir may be employed. In this method of nucleic acid composition loading, the orifice is contacted with the nucleic acid fluid composition under conditions sufficient for fluid to flow through the orifice and into the firing chamber of the head, where fluid flow is due, at least in part, to capillary forces. To assist in the flow of fluid into the orifice, back pressure in the form of suction (i.e. negative pressure) may be applied to the firing chamber (and reservoir, if present) of the head, where the back pressure will typically be at least about 5, and may be at least about 10 and even as great as about 100 inches of H2O or more.
To deposit nucleic acid onto the surface of a substrate according to the subject methods, the nucleic acid composition loaded thermal inkjet head is positioned in opposing relationship relative to the surface of the substrate, where the orifice is in opposition to the position on the substrate at which deposition of the nucleic acid is desired. The distance between the orifice and the substrate surface will not be so great that the volume of nucleic acid fluid cannot reach the substrate surface and produce a spot in a reproducible manner. As such, the distance between the orifice and the substrate surface will generally range from about 10 μm to 10 mm, usually from about 100 μm to 2 mm and more usually from about 200 μm to 1 mm.
After the head is placed into position relative to the substrate surface, the temperature of the heating element or resistor of the head is raised to a temperature sufficient to vaporize a portion of the fluid immediately adjacent to the resistor and produce a bubble. In raising the temperature of the heating element, the temperature of the heating element is raised to at least about 100� C., usually at least about 400� C. and more usually at least about 700� C., where the temperature may be raised as high as 1000 � C. or higher, but will usually be raised to a temperature that does not exceed about 2000� C. and more usually does not exceed about 1500� C. As such, a sufficient amount of energy will be delivered to the resistor to produce the requisite temperature rise, where the amount of energy will generally range from about 1.0 to 100 μJ, usually from about 1.5 to 15 μJ. The portion of fluid in the firing chamber that is vaporized will be sufficient to produce a bubble in the firing chamber of sufficient volume to force an amount of liquid out of the orifice.
The formation of the bubble in the firing chamber traps a portion or volume of the fluid present in the firing chamber between the heating element and the orifice and forces an amount or volume of fluid out of the orifice at high speed. The amount or volume of fluid that is forced out of the firing chamber can be controlled depending on the specific amount of nucleic acid that is desired to be deposited on the substrate. As is known in the art, the amount of fluid that is expelled can be controlled by changing one or more of a number of different parameters of the ink jet head, including: the orifice diameter, the orifice length (depth), the size of the firing chamber, the size of the heating element, and the like. Such variations are well known to those of skill in the art. As such, the amount or volume of fluid that is forced out or expelled from the firing chamber may range from about 0.1 to 1000 pl, usually from about 0.5 to 500 pl and more usually from about 1.0 to 250 pl. The speed at which the fluid is expelled from the firing chamber is at least about 1 m/s, usually at least about 10 m/s and may be as great as about 20 m/s or greater.
Upon actuation of the thermal inkjet head, as described above, fluid is expelled from the orifice and travels to the substrate surface, where it forms a spot on the substrate surface. In this manner, the nucleic acid is deposited on the substrate surface. As mentioned above, by varying the design parameters of the thermal inkjet head, the spot dimensions can be controlled such that spots of various sizes can be produced. In addition, multiple fires or patterning can be used to form larger features. With the subjects methods, one can produce spot sizes which have diameters ranging from a minimum of about 10 μm to a maximum of about 1.0 m. In those embodiments where very small spot sizes are desired, one can produce small spots that have a diameter ranging from about 1.0 μm to 1.0 mm, usually from about 5.0 μm to 500 μm and more usually from about 10 μm to 200 μm.
An important feature of the subject invention is that the deposited nucleic acid is capable of hybridizing to complementary nucleic acids. In other words, the deposition process does not adversely affect the nucleic acid of the sample, e.g. does not physically alter the nature of the nucleic acid such that it cannot subsequently participate in Watson-Crick type hydrogen bonding interactions.
The subject methods can be used to deposit a single nucleic acid sample on a substrate surface, i.e. a single nucleic acid spot, or a plurality of nucleic acid samples or spots on the surface, e.g. a pattern of spots as found in a nucleic acid array, where by plurality is meant at least 2, usually at least 5 and more usually at least 10. Where a pattern of spots is deposited on a substrate surface, the pattern may vary as desired. As such, the pattern may be in the form of organized rows and columns of spots, e.g. a grid of spots, across the substrate surface, a series of curvilinear rows across the substrate surface, e.g. a series of concentric circles or semi-circles of spots, and the like.
The subject methods and devices may be used to deposit nucleic acid samples on a variety of different substrates, including both flexible and rigid substrates. By flexible is meant that the support is capable of being bent, folded or similarly manipulated without breakage. Examples of solid materials which are flexible solid supports with respect to the present invention include membranes, flexible plastic films, and the like. By rigid is meant that the support is solid and does not readily bend, i.e. the support is not flexible. As such, rigid substrates are sufficient to provide physical support and structure to the nucleic acid spots present thereon. Furthermore, when the rigid supports of the subject invention are bent, they are prone to breakage.
The substrates may take a variety of configurations ranging from simple to complex. Thus, the substrate could have an overall slide or plate configuration, such as a rectangular, square or disc configuration. In many embodiments, the substrate will have a rectangular cross-sectional shape, having a length of from about 4 mm to 200 mm, usually from about 4 to 150 mm and more usually from about 4 to 125 mm and a width of from about 4 mm to 200 mm, usually from about 4 mm to 120 mm and more usually from about 4 mm to 80 mm, and a thickness of from about 0.01 mm to 5.0 mm, usually from about 0.1 mm to 2 mm and more usually from about 0.2 to 1 mm.
The substrates may be fabricated from a variety of materials. In certain embodiments, e.g. where one is interested in the production of nucleic acid arrays for use in research and related applications, the materials from which the substrate may be fabricated should ideally exhibit a low level of non-specific binding during hybridization events. In many situations, it will also be preferable to employ a material that is transparent to visible and/or UV light. For flexible substrates, materials of interest include: nylon, both modified and unmodified, nitrocellulose, polypropylene, polyester films, such as polyethylene terephthalate, and the like, where a nylon membrane, as well as derivatives thereof, is of particular interest in this embodiment. For rigid substrates, specific materials of interest include: silicon; glass; plastics, e.g. polytetrafluoroethylene, polypropylene, polystyrene, polycarbonate, and blends thereof, and the like; metals, e.g. gold, platinum, and the like; etc.
The substrate surface onto which the nucleic acid composition is deposited may be smooth or substantially planar, or have irregularities, such as depressions or elevations, or have a porous surface, such as is found in porous glass or silica. The surface may be modified with one or more different layers of compounds that serve to modify the properties of the surface in a desirable manner. Such modification layers, when present, will generally range in thickness from a monomolecular thickness to about 1 mm, usually from a monomolecular thickness to about 0.1 mm and more usually from a monomolecular thickness to about 0.001 mm. Modification layers of interest include: inorganic and organic layers such as metals, metal oxides, conformal silica or glass coatings, polymers, small organic molecules and the like. Polymeric layers of interest include layers of: peptides, proteins, polynucleic acids or mimetics thereof, e.g. peptide nucleic acids and the like; polysaccharides, phospholipids, polyurethanes, polyesters, polycarbonates, polyureas, polyamides, polyethyleneamines, polyarylene sulfides, polysiloxanes, polyimides, polyacetates, and the like, where the polymers may be hetero- or homopolymeric, and may or may not have separate functional moieties attached thereto, e.g. conjugated.
The methods find use in a variety of different applications in which it is desired to place or deposit a volume of a nucleic acid sample onto the surface of a substrate, where the nucleic acid is capable of participating in a hybridization event, e.g. is capable of bonding in a Watson-Crick base pair type fashion. Examples of applications in which the subject invention fmds use include: the preparation of diagnostic devices, e.g. test strips for nucleic acid analytes; the preparation of nucleic acid arrays; and the like, where the subject invention is particularly suited for use in the production of nucleic acid arrays, more particularly arrays of polynucleotides, e.g. cDNA arrays.
The subject invention may be used to produce nucleic acid arrays in which each spot on the array surface is small. As defined above, by small is meant that each spot on the array has a diameter that is at least about 1 μm, usually at least about 5 μm and more usually at least about 10 μm and does not exceed about 1 mm, usually does not exceed about 500 μm and more usually does not exceed about 200 μm. Because of the nature of the process, the nucleic acid on the surface of the array is capable of hydrogen bonding with complementary nucleic acids. Of particular interest in certain embodiments are arrays produced by the subject invention in which the nucleic acid is an oligonucleotide. Also of particular interest in many embodiments are arrays produced by the subject invention in which the nucleic acid is a polynucleotide, e.g. a cDNA.
Also provided by the subject invention are automated devices for depositing nucleic acids on a substrate surface, e.g. for producing nucleic acid arrays. The automated devices of the subject invention are analogous to conventional thermal inkjet printing devices, with the exception that the thermal inkjet head of the device is filled with a nucleic acid fluid instead of ink. Such automatic devices comprise at least a means for precisely controlling the position of the head with respect to a substrate and for firing the head. Such automated devices are well known to those of skill in the printing and document production art, and are disclosed in U.S. Pat. Nos. 5,772,829; 5,745,128; 5,736,998; 5,736,995; 5,726,690; 5,714,989; 5,682,188; 5,677,577; 5,642,142; 5,636,441; 5,635,968; 5,635,966; 5,595,785; 5,477,255; 5,434,606; 5,426,458; 5,350,616; 5,341,160; 5,300,958; 5,229,785; 5,187,500; 5,167,776; 5,159,353; 5,122,812; and 4,791,435; the disclosures of which are herein incorporated by reference.
Also provided by the subject invention are kits for use in depositing a nucleic acid on a substrate according to the subject methods. The subject kits include at least a thermal inkjet head and instructions for practicing the subject invention, where the instructions may be present as a package insert or on the packaging of the device. The subject kits may further include various reagents for preparing the nucleic acid fluid composition that is to be deposited on a substrate, e.g. reagents for practicing preparative PCR, such as polymerase, dNTPs, buffer medium and the like. In certain embodiments, the thermal inkjet head of the kit may be pre-filled with a nucleic acid fluid. In some embodiments, the kits may also comprise the substrate onto which the nucleic acid is to be deposited.
The following is offered by way of illustration and not by way of limitation.
A. Cy3 fluorescent dye end-labeled 1 kb and 0.1 kb double-stranded DNAs containing vector and hepatitis C virus sequences were generated by PCR using Cy3 end-labeled primers and a plasmid template containing hepatitis C virus sequences in the vector pSP64 (Promega, Madison, Wis.). Taq Gold enzyme (Perkin Elmer, Foster City, Calif.) was used for the PCR according to manufacturers instructions. The DNAs from the PCR reactions were concentrated using ethanol precipitation and re-suspended in 0.5 M sodium carbonate, pH9 at a concentration of 0.5 mg/ml. An HP 92261 A thermal ink jet head was used to fire 2500 droplets of 200 pl (total 0.5 μl) of each DNA onto parafilm. The material (0.5 μl) was collected and analyzed on a denaturing polyacrylamide gel to check for degradation. DNAs separated on the gels were visualized both with Stains-All (Sigma, St. Louis, Mo.) staining and fluorescence detection of the Cy3 label. The fluorescence scans were quantitated using ImageQuant software (from Molecular Dynamics, Sunnyvale, Calif.). The Cy3 scan of the 1 kb PCR product and associated primers is shown in FIG. 1A. Qualitatively, the DNA samples that were put into the inkjet looked the same as what came out, both for the first 0.5 μl (post-Jet-1) and the second 0.5 μl (post-Jet-2) samples. There were ˜20 mer oligos in the DNA samples (left-over PCR primers) that also look the same before and after inkjetting. The results demonstrate that most, if not substantially all, of the DNA seems intact in terms of breaks along the backbone of the DNA.
B. Gel electrophoresis experiments similar to those shown in 1. A above were performed to examine DNAs fired from an HP 51645 A thermal inkjet head (which has a smaller volume, higher velocity and greater fluidic sheer forces than the HP92261 A inkjet head). Native gel electrophoresis was employed to examine both 500 bp and 2200 bp DNAs. It was found that neither showed detectable degradation after firing from the inkjet. The results are depicted in FIG. 1B.
II. DNA samples were thermal inkjetted onto a glass substrate derivatized with aminopropyltrimethoxysilane-phenyldiisothiocyanate [Guo et al (1994) Nucl. Acids Res. 22:5456-65]. The DNA samples included a 314 bp hepatitis C virus sequence-containing PCR product prepared as described above and a 25 nt synthetic oligonucleotide prepared on an ABI synthesizer using standard phosphoramidite chemistry [Beaucage et al. (1981) Tetrahedron Lett. 22:1859-62; Matteucci et al (1981) J. Am. Chem. Soc. 103:3185]. The DNA samples were re-suspended at 0.5 mg/ml (314 bp PCR product) and 60 μM (25 mer) in 0.5 M sodium carbonate, pH 9.0. For each DNA, 1000-200 pl spots (total 0.2 μl) were jetted from an HP92261A thermal inkjet head in succession onto a defined position on the slide and identical samples of 0.2 μl were hand pipeted onto nearby positions on the same slide. Cy3-labeled oligonucleotide complementary to both the spotted oligos and PCR products was hybridized to the slides at a range of concentrations. Hybridization was detected by scanning for Cy3 fluorescence and quantitated using ImageQuant software. The results are provided in FIGS. 2A & 2B. Similar spotting and hybridization experiments were performed to examine DNAs fired from an HP51645 A thermal inkjet head. A 500 bp PCR product was fired from the inkjet and was found to have similar specific hybridization characteristics to hand-spotted DNA.
The results of these experiments show that the lower limit of detection and dynamic range of hybridization signal is virtually identical for the thermal inkjetted and hand-spotted oligos and PCR products indicating that the biological performance of the DNAs is not impaired by the thermal inkjetting process.
It is evident from the above results and discussion that a simple and efficient way to produce nucleic acid arrays is provided by the subject invention. By using the subject arrays, very small volumes of fluid can be deposited quickly on the substrate surface. Furthermore, the subject invention is a non-contact process such that the potential contamination and surface disruption that could arise from contact of an applicator with the substrate surface is eliminated. Finally, the thermal inkjet heads require less fluid to operate than comparable piezoelectric inkjet heads, thereby making the subject methods more economical than other methods of making arrays, particularly where expensive nucleic acid fluids are involved, e.g. cDNA compositions.
All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention.
Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it is readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.
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|Feb 25, 2014
|Hewlett-Packard Development Company, L.P.
|Fluid delivery system
|Oct 16, 2015
|Aug 2, 2016
|Twist Bioscience Corporation
|De novo synthesized gene libraries
|Aug 5, 2014
|Aug 9, 2016
|Twist Bioscience Corporation
|De novo synthesized gene libraries
|Jun 20, 2016
|Jan 31, 2017
|Twist Bioscience Corporation
|De novo synthesized gene libraries
|May 13, 2016
|Jun 13, 2017
|Twist Bioscience Corporation
|Compositions and methods for synthetic gene assembly
|Dec 12, 2002
|Jul 3, 2003
|Shogo Moriya
|Method for determining chum salmon haplotype using mitochondrial DNA
|Oct 8, 2002
|Jan 1, 2004
|Barth Phillip W
|Method and multiple reservoir apparatus for fabrication of biomolecular arrays
|Jul 10, 2002
|Jan 15, 2004
|Caren Michael P.
|Arrays with positioning control
|Sep 1, 2004
|Apr 28, 2005
|Caren Michael P.
|Method of performing array-based hybridization assays using thermal inkjet deposition of sample fluids
|Dec 15, 2004
|Aug 4, 2005
|Kirshner Brian M.
|Automated oligomer synthesis
|Mar 11, 2004
|Sep 15, 2005
|Surmodics, Inc.
|Array print buffers
|Mar 28, 2005
|Oct 6, 2005
|Kabushiki Kaisha Toshiba
|Encoded carrier and a method of monitoring an encoded carrier
|Feb 15, 2005
|Aug 17, 2006
|Dudenhoefer Christie L
|Method of forming a biological sensor
|Apr 29, 2005
|Nov 2, 2006
|Sarah Rosenstein
|Dispenser for making a lateral flow device
|Apr 29, 2005
|Nov 2, 2006
|Sarah Rosenstein
|Lateral flow device
|Nov 2, 2007
|Jun 5, 2008
|Sarah Rosenstein
|Lateral flow device
|Dec 19, 2006
|Dec 11, 2008
|Koninklijke Philips Electronics, N.V.
|Ink Jet Device for the Positioning of a Substance Onto a Substrate, Method for the Positioning of a Substance Onto a Substrate and Use of an Ink Jet Device
|Aug 13, 2007
|Feb 19, 2009
|Manish Giri
|Fluid delivery system
|May 16, 2007
|Aug 26, 2010
|Arcxis Biotechnologies, Inc.
|Pcr-free sample preparation and detection systems for high speed biologic analysis and identification
|Jul 8, 2009
|Mar 2, 2011
|Administraci�n General De La Communidad Aut�noma De Euskadi
|Methods for the diagnosis of multiple sclerosis based on its microRNA expression profiling
|Jul 15, 2011
|Jan 16, 2013
|Laboratorios Del. Dr. Esteve, S.A.
|Methods and reagents for efficient control of HIV progression
|Dec 19, 2006
|Jun 28, 2007
|Koninklijke Philips Electronics N.V.
|Ink jet device for the positioning of a substance onto a substrate, method for the positioning of a substance onto a substrate and use of an ink jet device
|Dec 19, 2006
|Oct 18, 2007
|Aleksey Kolesnychenko
|Ink jet device for the positioning of a substance onto a substrate, method for the positioning of a substance onto a substrate and use of an ink jet device
|Jul 8, 2010
|Jan 13, 2011
|Administraci�n General De La Comunidad Aut�noma De Euskadi
|Methods for the diagnosis of multiple sclerosis based on its microrna expression profiling
|Dec 20, 2012
|Jun 27, 2013
|Illumina, Inc.
|Methylation biomarkers for ovarian cancer
|Jun 12, 2013
|Dec 19, 2013
|Fundaci�n Centro Nacional De Investigaciones Cardiovasculares Carlos Iii
|Marker for determining the therapeutic potential of mesenchymal stem cells, an inhibitor of the marker for therapeutic use and stem cells with low expression of the marker for therapeutic use
|May 20, 2014
|Nov 27, 2014
|3P Biopharmaceuticals
|Alphaviral vectors and cell lines for producing recombinant proteins
|U.S. Classification
|506/16, 435/287.2, 536/25.3, 436/94, 536/23.1, 506/18, 506/40, 506/30, 506/17, 422/547, 435/6.1, 435/6.11
|International Classification
|Cooperative Classification
|European Classification
|Dec 24, 1998
Owner name: HEWLETT-PACKARD COMPANY, CALIFORNIA
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:BRUHN, LAURAKAY;CAREN, MICHAEL P.;SCHEMBRI, CAROL T.;ANDOTHERS;REEL/FRAME:009671/0733
Effective date: 19980908
|Apr 24, 2000
Owner name: HEWLETT-PACKARD COMPANY, COLORADO
Free format text: MERGER;ASSIGNOR:HEWLETT-PACKARD COMPANY;REEL/FRAME:010759/0049
Effective date: 19980520
|May 30, 2000
Owner name: AGILENT TECHNOLOGIES INC, CALIFORNIA
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:HEWLETT-PACKARD COMPANY;REEL/FRAME:010977/0540
Effective date: 19991101
|Apr 3, 2006
|Fee payment
Year of fee payment: 4
|Mar 18, 2010
|Fee payment
Year of fee payment: 8
|May 9, 2014
|Maintenance fee reminder mailed
|Oct 1, 2014
|Lapse for failure to pay maintenance fees
|Nov 18, 2014
|Expired due to failure to pay maintenance fee
Effective date: 20141001


text:
7 Tips for Big Data Success
Experienced Big Data users understand the paradox – Big data starts small. Rather than rush to implement new techniques and technology a small win can help to drive big results. Patience is often hard when there are big problems to solve, but creating a solid foundation for success can make things go faster when you need to implement Big Data transformations for analytics and dashboards. Our friends at Big Data Analytics News provide 7 quick tips to consider before you launch – start small and drive to big results.
The entire article is located on Big Data Analytics News
Related topics
– Agile Dashboards with Pentaho CTools
– Convenience Store Performance Analytics
Learn how business intelligence helps you identify profit leaks and achieve new goals.
"I always find value and inspiration from BandyWorks' newsletter!"
--John, CEO

subset: commoncrawl


text:
Monday, May 24, 2010
Friday, May 21, 2010
Oh my...
I haven't posted here in a long, long time. Well, Harper is all grown up now. She starts college next week, you know. Kidding, kidding, of course. It does feel like time is flying by.
Harper IS doing lots of cool things. She repeats almost everything you say and tries to sing along when we sing (poorly even). This morning we were singing, "Mama's don't let your babies grow up to be cowgirls." Harper always echoes the last word. Cowgirls!
She's quite the helper in the art studio now too. She likes to rearrange all the buttons for me. Oh, and occasionally dump them out on the floor for "inventory".
She likes to pick up the phone and "call" Grandma... and every other relative she can think of. Oh, and animals too. "Hello, Elephant!" "How you?" "Bye, Elephant!"
She is truly a joy. SO lucky. Ever so lucky.
And now for some interpretive dance to the music of Cake...
Posted by Shelly at 11:56 AM No comments:
Sunday, May 9, 2010
A Milestone
Miss Harper now calls that girl in the mirror Harper! Just a few days ago, she saw a photo of herself and said, "Harper!" and then pointed at herself and said "Harper!" She's sweeter and sweeter each day. I didn't think that was possible. I am the luckiest Mommy on the planet. :)
Posted by Shelly at 5:55 PM 2 comments:
Saturday, May 8, 2010
Flower Child
Grandpa Mark took this photo of Miss Harper yesterday... thought I'd share it with you. Happy Saturday.
Posted by Shelly at 5:46 AM 1 comment:
Friday, May 7, 2010
Harper is 2!
Harper is 2!, originally uploaded by shellypaints.
Posted by Shelly at 7:49 AM 1 comment:
Grandma's Cookies
Posted by Shelly at 7:43 AM 1 comment:
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text:
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Spring 2018 Japan Cherry Blossom Tour with Zorie
This will be an intimate visit with a small group of 10-15 people. Starting at the "must be seen to be believed" Robot Restaurant in Tokyo's Shinjuku, this is whirlwind tour through the sights, sounds and tastes that make Japan a cultural jewel, as well as seeing locations from the film. From the Kabuki theater of Ginza, to the Oedo Onsen hot springs, and the Zen gardens of Kyoto, as much a rare opportunity for newcomers to dive into the unique culture of Japan as it will be for seasoned travelers to revisit their favorite after hours ramen spots or stop by their old dojo. And of course - Cherry Blossoms!
For more info email email@example.com


text:
A short history of BrixtonJune 17th, 2007
A short history of Brixton
A fascinating little timeline of Brixton. From a roman road crossing, to upper middle class, to working class, to immigrants, and now an up and coming neighborhood back to some posh.
Posted in Asides | No Comments »


text:
Strawberry Ruby Grapefruit Preserves P&Q
Posted July 20, 2009on:
- In: 1
- 23 Comments
A P&Q on time – miracles do happen. Have fun with this one – will be fun to see how every chooses to use the preserves.
23 Responses to "Strawberry Ruby Grapefruit Preserves P&Q"
When I made this I just used grapefruit juice and zest, and I can't imagine doing it differently! It was pretty tart, so I think you're supposed to use the grapefruit in place of the lemon/orange.
I plan on making these as a filling for some vanilla cupcakes. I'm still debating whether or not to make theme strawberry cupcakes with a kiwi frosting. I guess I'll see =]
Actually looking it over, I may just make strawberry cupcakes and a regular buttercream frosting =]
Thanks for the tip on the generous recipe size; I was planning on making a whole recipe but maybe I'll scale it back a bit. It's my week hosting and I planned to make the preserves over the weekend, but I got too busy.
Oh NIna that is a wonderful idea. I can't wait to see what you come up with. Im going on a little weekend trip this week and may not get to this. I will try my best though!
Thinking of making the strawberry-rhubarb version instead (lots of unused rhubarb in my garden). That would be OK, wouldn't it?
Hanaa Strawberry and Rhubara go well together. My friend makes that kind of pie and it is great. Im sure it would be great in preserves!
I decided to sit this one out. We are leaving Friday and Im not going to have time to do this recipe. Sorry!
I didn't realize it until I chose this recipe, but each type of preserve in the book is regarded as a separate recipe. That version sounds good too, though!
I halved the recipe for this one! I still have a tonne of jam from my last creation. My house smells so yummy!
Katie xox
I finally made the both the preserves and cupcakes. and 1/4 of the recipe fit perfectly into my 24 cupcakes! No leftovers!
I halved it too Katie and completely echo your sentiments on the scent – it is phenomenal!!
Are we going to get the list of Aug. recipes soon? Im trying to plan ahead as Aug. is a busy month!
When I made this, we all thought the house smelled like a candy factory or something. We had the windows open and even the backyard smelled delicious!
I just finished making Strawberry Rhubarb preserves (sans orange zest). It is delicious. If you're thinking of skipping this week's recipe, don't! I made 1/4 of the recipe and it took me 40 min from start to finish. After the mixture reached 212 F, it only 15 more minutes to get to the right thickness. Make it, you won't be sorry.
Forgot to mention… I just had it on toast (I know, it's 9:00pm). Yum!
I agree. I need to get my ingredients much ahead of time since I dont have time during the week to get them. If they could be put up before the end of this weekend that would be great!
I made it last night; I did the whole recipe and canned it. I don't know what you would do with an entire batch of this if you didn't can it – it really makes a lot!
Does anyone have tips about auto-posting on blogger? From what I understand, you change the date and time under "post options," hit publish post when you're done, and then it becomes "scheduled" until the selected time when it auto-posts. It's worked twice for me and every other time I've tried, my post has remained in my post list as "scheduled." I'm running a half-marathon tomorrow morning, so I need to go to bed and get up early (and not be worried about whether or not my post published).
Hi friends!
I thought that I had everything I needed to make this, but I looked in the fridge and no apple! oops! I will make this at some point this week, and then I'll do a double post next week.
I wasn't going to participate this week but soooo glad that I did. The preserves are delicious. Thanks for all the helpful tips before I got started. I guess that is a good thing about getting started late.
Comments are closed.


text:
Clayton State University
Master of Science in Criminal Justice
College of Arts & Sciences
2000 Clayton State Boulevard
Morrow, GA 30260-0285
(678) 466-4855
(678) 466-4899 (fax)
Dr. Jason Davis
College of Arts & Sciences
Hello and welcome to the Criminal Justice Master's Program,
The Clayton State School of Graduate Studies offers a Master of Science in Criminal Justice degree that includes two distinct academic tracks: Administration of Justice and Criminology, Law, & Society. The Administration of Justice track is a 33 hour program that emphasizes a comprehensive understanding of crime and criminal justice research, policy planning, and criminal justice administration. This track is especially recommended for current and aspiring practitioners in the criminal justice and social services communities. The Criminology, Law, & Society track is a 33 hour program designed for students who desire more advanced preparation for doctoral studies in criminology or criminal justice. This track is also intended for students interested in pursuing careers as criminal justice research or policy analysts.
The curricula includes a sequence of required courses that focus on crime typologies, legal institutions, the criminal justice system, research methodology, program evaluation, and criminological theory. Students will also be required to conduct independent research that critically analyzes a criminal justice policy or topic. Regardless of which track a student pursues, both concentrations will equip students with the research, communication, and analytical skills necessary for leadership positions in the justice and law enforcement professions or admission in competitive criminal justice, criminology, law, or related programs.
Students in the program will be strongly encouraged to work directly with Criminal Justice faculty as well as faculty representing other disciplines such as Sociology, Political Science, and Psychology. The Criminal Justice faculty hold degrees from nationally recognized graduate programs and boast both practical experience and scholarly productive particularly in the areas of corrections; law enforcement; drug enforcement, distribution, and policy; white collar crime; criminological theory; gangs; and intersectionality of crime including race, social class, and gender.
Mailing address: School of Graduate Studies
Clayton State University
2000 Clayton State Blvd
Suite 133 Faculty Hall
Morrow, GA 30260
(678) 466-4113
Email address: firstname.lastname@example.com
Web address: www.clayton.edu/graduate


text:
Antalya is situated in the south western coastline of Turkey with the Mediterranean and Aegean Seas lapping gently to its coastline. The region is a popular tourist resort with resorts that include Alanya, Antalya, Belek, Colakli, Incekum, Kemer, Manavgat, Side and Tekirova. It has a superb climate, fantastic views of the idyllic coastline and it also has many historical attractions. Coupled with its natural beauty, what more could you ask from a holiday destination?
The beaches are a popular attraction and Antalya is often referred to as the Turkish Riviera for its long stretches of sandy beaches, sunshine and excellent golfing opportunities.
The region has an abundance of mountains and valleys: The Taurus Mountain range is one which is absolutely enchanting, particularly in the winter when the mountains are snow capped. There are 3 National Parks in Antalya where you will find a back to basic feel, with all the charm and unusual flora which is unique to the Antalya Region. The National Parks are the Coprulu Canyon, Termessos National Park and the Olympus National Park. If you are a nature lover, you will find plenty of places to spend some time enjoying local serenity and peace.
The hospitality of the local people is friendly; they can't do enough to help you. There are plenty of restaurants which serve an amazing mix of traditional Turkish food to something a little more international. Eating out is not expensive and you will find families are welcomed, with value for money meals.
It has a typical Mediterranean climate with long, hot sunny days during the summer and sea temperatures between May to November rarely go below 20°C. There are cooling breezes from the north-east which help to keep the summer temperatures more bearable.
Tourism is the main economy for Antalya, although it is also agricultural and is, in fact, Turkey's largest producer of citrus fruits, flowers and bananas for export.
Antalya is the venue for many events and festivals throughout the year including the popular Golden Orange Film Festival which is held in September. Many of the local celebrations are religious as Turkey is mainly Muslim. For that reason we would advise clothing to be worn appropriately, particularly ladies when off of the beach and especially in any religious buildings.
A holiday in Antalya will provide you with historical sights, fantastic scenery, thrills, excitement, water sports, diving, snorkelling, golf, mountain walks and some of the world's finest beaches. There are heaps to do for families and couples alike. You can find a variety of budget, low cost and value for money hotels which won't break your budget.
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text:
Finzelberg GmbH and Co. KG
Finzelberg GmbH and Co. KG
Obolskiy D.,University of London |
Pischel I.,University of London |
Pischel I.,Phytolab GmbH and Company KG |
Feistel B.,Finzelberg GmbH and Co. KG |
And 3 more authors.
Journal of Agricultural and Food Chemistry | Year: 2011
Artemisia dracunculus L. (tarragon) has a long history of use as a spice and remedy. Two well-described "cultivars" (Russian and French) are used widely and differ in ploidy level, morphology, and chemistry. Key biologically active secondary metabolites are essential oils (0.15-3.1%), coumarins (>1%), flavonoids, and phenolcarbonic acids. In vivo studies mainly in rodents, particularly from Russian sources, highlight potential anti-inflammatory, hepatoprotective, and antihyperglycemic effects. Despite concerns about the toxic effects of two of its main constituents, estragole (up to 82%) and methyleugenol (up to 39%), no acute toxicity or mutagenic activity has been reported at doses relevant for human consumption. Water extracts of A. dracunculus contain very low amounts of estragole and methyleugenol and, therefore, are considered to pose a very limited risk. Overall, a stronger focus on clinical studies and precise taxonomic and phytochemical definition of the source material will be essential for future research efforts. © 2011 American Chemical Society.
Begrow F.,University of Munster |
Engelbertz J.,University of Munster |
Feistel B.,Finzelberg GmbH and Co. KG |
Lehnfeld R.,Phytolab GmbH and Co. KG |
And 2 more authors.
Planta Medica | Year: 2010
Thyme is a herb with broncholytic und secretomotoric effects. Its activity on receptors as a possible mechanism of action was demonstrated. Major components are thymol and carvacrol which are claimed to be responsible for its effects and, therefore, used for standardization in the German pharmacopoeia (0.03% phenols calculated as thymol). Our aim was to investigate the impact of thymol by using thyme extracts with either normal or extremely low thymol concentrations (<0.005% or >0.038%). The antispasmodic effect on smooth muscles of the trachea and the ileum and the effect on ciliary activity (respiratory clearance) were investigated. In addition, pure thymol and carvacrol were investigated separately and in spiking experiments. Thymol and carvacrol had a concentration-dependent antispasmodic effect in the rat trachea either being stimulated by acetylcholine, K+ or Ba++. The same result was observed with respect to the increase of mucociliary transport in mice. Extracts with very low thymol contents are effective in all models used except acetylcholine-induced rat ileum contraction. When thyme extracts with normal thymol contents or with very low thymol contents were compared, the extract with normal thymol contents was more effective, both as a relaxant (rat ileum) and as an antispasmodic compound (rat trachea contraction induced by either acetylcholine, Ba++ or K+) and in ciliary transport experiments. Thyme extracts with very low thymol contents (practically free of volatile oil) were equally effective with respect to endothelin effects. When an extract with very low thymol contents is spiked with increasing concentrations of thymol, a concentration-dependent increase concerning the antispasmodic effect (Ba++-induced trachea contraction) is observed. In conclusion, the data show that in various models of antispasmodic effect (ileum and trachea) and by measuring ciliary activity, thymol (and carvacrol) is (are) active, although other not identified components of thyme extract appear to be very important as well, since extracts with very low thymol contents are active. On the basis of these results the standardization on thymol alone appears not to be justified. © Georg Thieme Verlag KG Stuttgart New York.
Godard M.P.,Western Illinois University |
Ewing B.A.,Western Illinois University |
Pischel I.,PhytoLab GmbH and Co. KG |
Ziegler A.,PhytoLab GmbH and Co. KG |
And 2 more authors.
Journal of Ethnopharmacology | Year: 2010
Aim of the study: The aim of this study was to evaluate the acute and chronic effects of OpunDia™ (Opuntia ficus-indica) in obese pre-diabetic men and women. Materials and methods: This double-blind placebo controlled study included participants (age range of 20-50 years) randomly assigned to one of the two groups and given a 16-week supply of either the 200. mg OpunDia™ (n= 15), or placebo (n= 14). The acute phase of the study consisted of an oral glucose tolerance test (OGTT) with a 400. mg bolus of OpunDia™ given 30. min before orally ingesting a 75. g glucose drink. Baseline and post 16-week concentrations of glucose, insulin, hsCRP, adiponectin, proinsulin, Hb1Ac, cholesterol, and a comprehensive metabolic panel were collected along with body composition measured via densitometry (BOD POD). A repeated measures ANOVA was conducted to determine any significant interactions between group and time. Follow-up analysis was performed to determine differences among groups at each time point. Paired t-tests were performed on all variables to determine if any within group differences existed across time. Results: There was a statistically significant decrease (P< 0.05) in the blood glucose concentrations at the 60 (205.92 ± 36.90 and 188.84 ± 38.43. mg/dL, respectively), 90 (184.55 ± 33.67 and 169.74 ± 35.16. mg/dL, respectively) and 120. min (159.24 ± 17.85 and 148.89 ± 24.86. mg/dL, respectively) time points with the pre-OGTT compared to the OpunDia™ bolus trial. There were no between-group differences found with the OGTT time points, area under the curve, blood chemistry variables (insulin, hsCRP, adiponectin, proinsulin, Hb1Ac), diet analysis variables (carbohydrates, fat, protein and total kcals), body composition variables (fat mass, fat free mass, percent body fat and total body weight), or blood chemistry safety parameters (comprehensive metabolic panel) pre-to-post 16-week intervention. Conclusions: This study shows the acute blood glucose lowering effects and the long-term safety of the proprietary product OpunDia™, thus supporting the traditional use of Opuntia ficus-indica for blood glucose management. © 2010 Elsevier Ireland Ltd.
Algieri F.,University of Granada |
Rodriguez-Nogales A.,University of Granada |
Garrido-Mesa N.,University of Granada |
Zorrilla P.,University of Granada |
And 8 more authors.
Journal of Crohn's and Colitis | Year: 2014
Nowadays, there is an increasing interest for alternative options in the treatment of inflammatory bowel diseases (IBDs) that combine efficacy and an adequate safety profile. Methods: The intestinal anti-inflammatory effects of Serpylli herba, the officinal drug in the European Pharmacopeia composed by the aerial parts of wild thyme (Thymus serpyllum), were evaluated in the trinitrobenzenesulfonic acid (TNBS)-induced rat colitis and dextran sodium sulfate (DSS)-induced mouse colitis, which are well characterized experimental models with some resemblance to human IBD. Results: S. herba extract exerted an intestinal anti-inflammatory effect in both experimental models of colitis, as evidenced both histologically, since it facilitated the tissue recovery of the damaged colon, and biochemically as showed by the improvement of the different inflammatory markers evaluated, including myeloperoxidase activity, glutathione content, and leukotriene B4 levels as well as the expression of the inducible proteins iNOS and COX-2. This beneficial effect was associated with the reduction in the expression of different cytokines, like TNFα, IL-1β, IFNγ, IL-6 and IL-17, the chemokine MCP-1, and the adhesion molecule ICAM-1, thus ameliorating the altered immune response associated with the colonic inflammation. Conclusion: S. herba extract displays an anti-inflammatory effect on different models of rodent colitis that could be attributed to its immunomodulatory properties. © 2013 European Crohn's and Colitis Organisation.
Walstab J.,University of Heidelberg |
Kruger D.,TU Munich |
Stark T.,TU Munich |
Hofmann T.,TU Munich |
And 5 more authors.
Neurogastroenterology and Motility | Year: 2013
Background: Beneficial effects of ginger in the treatment of gastrointestinal (GI) problems and chemotherapy-induced nausea and vomiting are well accepted. In rodents, the action of ginger seems to be mediated by the inhibition of 5-HT3 receptors, which are established targets to combat emesis and irritable bowel syndrome. Methods: Heterologously expressed human 5-HT3A or 5-HT3AB receptors were characterized by means of Ca2+influx studies using HEK293 cells. Complementing Ca2+ measurements in Fluo-4-AM-stained whole-mount preparations of the human submucous plexus were carried out. Furthermore, [3H]GR65630 binding assays were performed to reveal the mode of action of ginger and its pungent compounds. Key Results: We show for the first time that ginger extracts and its pungent arylalkane constituents concentration-dependently inhibit activation of human 5-HT3 receptors. Ginger extracts inhibited both receptors with increasing content of pungent compounds, confirming that these are part of ginger's active principle. Inhibition potencies of the arylalkanes 6-gingerol and 6-shogaol on both receptors were in the low micromolar range. A lipophilic ginger extract and 6-gingerol had no influence on 5-HT potency, but reduced the 5-HT maximum effect, indicating non-competitive inhibition. The non-competitive action was confirmed by [3H]GR65630 binding, showing that the ginger extract did not displace the radioligand from 5-HT3A and 5-HT3AB receptors. The potential relevance of the inhibitory action of ginger on native 5-HT3 receptors in the gut was confirmed in whole-mount preparations of the human submucous plexus. While a general neurotoxic effect of 6-gingerol was ruled out, it inhibited the 2-methyl-5-HT-mediated activation of 5-HT3 receptors residing on enteric neurons. Conclusions & Inferences: Our findings may encourage the use of ginger extracts to alleviate nausea in cancer patients receiving chemotherapy and to treat functional GI disorders. © 2013 Blackwell Publishing Ltd.
Finzelberg GmbH and Co. KG | Date: 2011-08-26
The invention relates to aboveground plant parts of Sideritis ssp. or extracts produced therefrom for use to boost cognitive performance.
Finzelberg GmbH and Co. KG | Date: 2014-04-17
Finzelberg GmbH and Co. KG | Date: 2015-03-12
The invention relates to aboveground plant parts of Sideritis ssp. or extracts produced therefrom for use to boost cognitive performance.
Finzelberg GmbH and Co. KG | Date: 2013-06-10
Finzelberg GmbH and Co. KG | Date: 2013-12-03


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Many government service contractors successfully enter the government market with relatively straightforward T&M (Labor Hour) contracts. Eventually an opportunity to bid and perform larger and more complex cost reimbursement contracts will present itself. The skills required to manage T&M contracts differ markedly from the skills required to manage a cost type contract. While the statements of work may be very similar, the administrative requirements associated with managing a cost type contract greatly exceed those required for T&M work. Failure to comply with these requirements can have serious ramifications.
This 90- minute webinar will help you gain an understanding of your obligations… …read more
Read more here: Events


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Banking & Finance Law involves individuals and businesses in transactions with federal and state-chartered financial institutions -- including issues related to bank accounts, negotiable instruments, loans, interest rates, regulatory compliance, taxes, and more. Banking and finance law applies to those individuals and institutions that lend or borrow money. Lenders typically include banks, leasing companies, finance companies and other financial institutions. Borrowers are individuals, corporations, institutions or the government.
Williamson is an Upstate New York town in the northwest part of Wayne County, New York, U.S. on the south shore of Lake Ontario. The population was 6,777 at the time of the 2000 census. The town is named after Charles Williamson, a land agent of the Pultney Estate. Its ZIP code is 14589, and telephone exchanges 589 and 904 in area code 315.
Banking And Finance Law Lawyers In Williamson New York


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Lewisboro is a town in Westchester County, New York. The town is named after John Lewis, an early settler.
South Salem boasts an incredible public transit system so those who live further out can easily get to where they need to.
For those looking for a sense of stability South Salem is just the place to settle down in due to the high percentage of residents who own their properties. When it comes to the populations in this area, you wont be shocked to find that there is a high percentage of people who are college-educated. You'll be able to enjoy good company at dinner parties if you move to this area, as this is a popular area for married couples to set up house. In terms of the wealth of residents in South Salem, their incomes tend to be higher than average.
The average home listing price in South Salem last year was $603,151 - a small increase of 1.6 percent from last year. Meanwhile, the number of homes on the market modestly increased by 4.2 percent year-over-year to reach 74 listings. A number of properties in South Salem feature beautiful swimming pools which help mitigate the heat of the summer and allow for loads of fun for the entire family. This town's ambiance is defined by the scattering of colonial-style homes which lend sophistication to the area.
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Angus Taylor, Federal Member for Hume | Sydney Morning Herald | 29 September 2015
Australia's media ownership rules come from a pre-internet era and in the face of massive technological changes, there needs to be a complete overhaul.
This year, for the first time, I followed the Tour de France on my phone, not on TV. As a sports nut, I thought it was brilliant: free-to-air television couldn't compete with the availability, information and commentary from the Tour de France app. For me, this said it all about the new disruptive changes bearing down on traditional free-to-air television. It also told a story about the urgent need to modernise and streamline our media legislation. It is long past time for change.
We have all watched video kill the radio star, downloads kill off CDs, Amazon knock off many bookstores and the internet's effect on newspapers. Similar forces are driving massive changes to traditional TV, which is now facing challenges from competitors such as Netflix, clever sports codes and online news streaming.
We need to change the rule book so that the new digital era can flourish and at the same time make sure we don't go too far.The Broadcasting Services Act (1992) is from a pre-internet era. It restricts broadcasters to a reach of less than 75 per cent of the population (the "reach rule"). Yet now, any broadcaster can reach the entire population through the internet. For example, Channel 7 reaches all of Australia through its streaming service, Plus7, as does the ABC with iView.
As a regional MP I despair at the impact of the reach rule. It is squeezing the life out of our regional TV networks. As the metropolitan networks sell them content at high prices, they cut local news content to the bone, and cut new investment in transmission towers. They have no choice.
TV towers might seem unnecessary in the modern era, but free-to-air will remain important for older viewers and for high-profile national TV events. To make things worse, large parts of regional Australia lost TV reception in the recent switchover from analogue to digital. Despite the former Labor government spending more than $900 million on the switchover, not a single new tower was funded.
Broadcasters also face restrictions in merging with radio and newspapers. The "two out of three" rule prevents ownership of TV, radio and newspapers in any one licence area. Again, in a modern era the obvious answer is to combine written content with TV and radio, just as the ABC does. In practice, this rule means that a newspaper company like Fairfax can't add both radio and TV to its network.
In the past, this restriction may have been justifiable. Media markets were restricted to a single town or city, and there was little "convergence" across media channels.
But now we can view video, listen to podcasts or live streams and read news articles on a single website, and there is fierce competition crossing traditional geographic boundaries. Modern journalists, particularly in regional areas, are adept at capturing video and audio while writing articles and tweeting.
The answer, as many already know, is to get rid of this anachronistic legislation. In the process it will be possible to manage a sensible transition where local news content and TV transmission is protected.
In practice, this means repealing the reach rule, the two-out-of-three rule and streamlining a number of other restrictions. One of the trickier restrictions is anti-siphoning, preventing pay TV from putting our most loved sporting events behind their paywalls. Yet, in time sporting codes are likely to take control of their own product by streaming online, as the Tour de France has already done (albeit with the co-operation of local networks). Online they are essentially free from regulation. Anti-siphoning rules will become increasingly redundant as content moves online.
Uber (and its customers) is single-handedly deregulating a taxi industry, which excelled in tying itself up in red tape. Similarly, AirBnB is shaking up the accommodation industry. While complete deregulation probably won't be the right result, we need a very significant reset that reflects the way we now receive information and the way we will receive it in the future.
Media deregulation has been waiting in the wings for far too long. If we don't act quickly to ensure a smooth transition, large parts of our media sector are at risk in the same way the traditional music industry, bookstores and taxis are at risk. Local news content, TV transmission and access to major sporting events are also at risk.
Angus Taylor is the federal member for Hume. He was previously a partner at management consultants McKinsey & Co and Port Jackson partners.
View the article on The Sydney Morning Herald.


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Enjoy the beautiful weather this weekend with a trip to the zoo or a hike through a park!
5 Saturday
Beatrix Potter's Furry Friends at Tippecanoe: Celebrate Beatrix Potter's 150th birthday! We'll share stories, author facts, and learn more about animals like the ones she illustrated. Meet a variety of live animals with the chance to get up close!
Tippecanoe Library, 3912 S. Howell Ave., Milwaukee. 2 to 3:30 p.m. mpl.org
"The Big Blue Ball": The Lake Country Children's Theater presents the play performed by Sunset Playhouse Bug in a Rug.
Arrowhead High School North Campus Theater, 800 North Ave., Hartland. 1:30 p.m. $6/person. lakecountrychildrenstheater.org
Christmas Fantasy House: Your chance to visit a home decorated for the holidays by local businesses.
Delafield. $20/Adult, Free/children 12 and under. Proceeds go to support the Ronald McDonald House.
Family Free Day at the Zoo: If you want to stay warm, we have many indoor animal exhibits for your enjoyment. If you're more of the outdoorsy type, you'll also find many outdoor animal exhibits.
Milwaukee County Zoo, 10001 W. Bluemound Rd., Milwaukee. 9:30 a.m. to 4:30 p.m. milwaukeezoo.org
Family Nature Hike: Deer Tracking: Spend an hour with a naturalist on a family-friendly hike. White-tailed deer are on the move, making it easier to find the signs they leave behind. We may even see a real deer!
Havenwoods State Forest, 6141 N. Hopkins St., Milwaukee. 10:30 to 11:30 a.m. friendsofhavenwoods.org
Glacial Ghosts: Ice Age History of Whitnall Park: The Ice Age lives on! Join us for a hike back 15,000 years to an era when glaciers were in retreat and mammoths roamed. Discover how glaciers left their mark to create the Whitnall Park we enjoy today. Be prepared to hike over uneven terrain and possible muddy trails.
Wehr Nature Center, 9701 W. College Ave., Franklin. 9 to 11 a.m. $10/person, $7/Milwaukee County resident, $5/Friends of Wehr members; registration required by November 3. friendsofwehr.org
"Mole Hill Stories": First Stage, in collaboration with Danceworks, performs three of Lois Ehlert's most famous stories. Part of First Stage's First Step series, designed to provide families and younger children (ages 3 to 7) an introduction to live theater.
Milwaukee Youth Arts Center, 325 W. Walnut St., Milwaukee. Runs through November 20, Check website for times. $14/person. firststage.org
6 Sunday
Christmas Fantasy House: See November 5 listing.
"Mole Hill Stories": See November 5 listing.


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Christian Baptism
Even before the time of Jesus, many mystery religions of the ancient Mediterranean had some sort of ceremony that people had to do in order to join the cult. Some of these, like the Eleusinian mysteries, involved dipping yourself in water to wash away your past life, and to show that you were beginning a new life.
Christianity, from the very beginning, also had this idea of dipping yourself in water, which is called baptism. Before Jesus began preaching, he had himself dipped in water by his cousin, John, who is therefore called John the Baptist.
During the Roman Empire, Christians, like other people, were usually baptized as adults. That way they were old enough to understand what they were doing. In fact, people often waited to be baptized until they were dying. If being baptized washed away all your sins, so you could go to Heaven, people wanted to wait until the last minute so that they would be sure of going to Heaven. If you got baptized earlier, you might do something bad, and not go to Heaven after all. The emperor Constantine, for instance, was not baptized until he was dying, in 337 AD.
But later on, in the Middle Ages, people began to worry about dying by accident, quickly, without having time to get baptized. Then people started baptizing their babies, to be sure of getting it done in case the babies died.
Some famous baptisteries (buildings where people got baptized) are in Florence and in Pisa.
Learn by doing: were you baptized? why or why not?
More about John the Baptist
Bibliography and further reading about the history of baptism:


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We have incredible contributions from: Mattie Weiss, Wellstone Action and Erik Peterson, Wellstone Action; Rishi Awatramani, Virginia New Majority; Charles Lenchner, Organizing 2.0; and Jessamyn Sabbag, Oakland Rising.
Mattie Weiss, the director of Campus Camp Wellstone (a program of Wellstone Action) is a long-time youth movement organizer, writer, and leader. Mattie wrote two chapters of the book, How to Get Stupid White Men Out of Office, which she toured around the country, organizing and speaking on behalf of the League of Pissed Off Voters in the 2004 presidential election.
Erik Peterson has 25 years of experience as a community-based educator, trainer, and community and electoral organizer. He has served at all levels of campaign organizing in state and local races, most recently as the lead consultant for Mark Ritchie's successful 2006 campaign for Minnesota Secretary of State, and as the northern Minnesota Get-Out-the-Vote (GOTV) director for America Votes.
Building Power
"Electoral politics without community organizing is a politics without a base. And community organizing without grassroots electoral politics is a marginal politics. And electoral politics and community organizing without good progressive policy is a politics without a head – without a goal." - Senator Paul Wellstone
Wellstone Action is focused on building long-term, strategic progressive/Left power and enacting strong, resource-distributive, progressive public policy. We do this work within a framework we call, "The Wellstone Triangle."
Grassroots Organizing
In one corner of the triangle we have grassroots organizing (encompassing community, identity-based, and labor organizing), where we grow our organizations and movements. This is the work of building relationships and trust within communities; finding common ground that ties our issues together so our collective efforts magnify each other; building commitment and infrastructure around a compelling vision; and recruiting, training, mentoring, and supporting new leaders.
Electoral Politics
Another corner of the triangle represents electoral campaigns, in which we elect decision-makers committed to our agenda and accountable to our communities. It involves investing in candidate recruitment and development with a long-term strategy for moving good candidates toward higher office; and investing in training a new generation of grassroots political campaign organizers.
Public Policy
The third component of the Wellstone Triangle is about setting an agenda. Ideas inspire us; values ground and center us; public policies are how we enact our ideas and values in the real world. Moving policy is not just about drafting good legislation. This is the place of idea work, where we develop strategies to shift values and debates at the level of mass consciousness. We also develop the new generation of intellectuals and policy writers who are connected to our two other corners of grassroots organizing and electoral politics.
Historically, progressives and Left organizers within each of these three corners of the triangle have operated in silos, away from and even disdainful of one another. This has seriously weakened us. For example, over this decade young people have gotten more powerful in their capacity to mobilize around elections. We were the heart and many of the limbs of the Obama campaign. But now that our candidate is in office and the battle over health care, war, civil rights and immigration is going down, our voices are noticeably absent. While we were building our capacity to work on elections we developed precious little experience mobilizing around local, state and national policy, such that the man we put in office has no reason to be accountable to us.
Similarly, policy and decision-makers without a grassroots movement of people behind them are frequently either ineffectual or create policy that is damaging to our communities (intentionally or not). At a training we did with prostituted women several months ago, a sympathetic state senator came to talk about the anti-trafficking legislation she had authored. She is a strong supporter of the rights of sex workers and has the capacity to move ideas into law, but she had drafted the legislation without the voices and certainly without the mobilization of those directly impacted by the policy. When the women sat down with the language of the bill, they immediately identified ways it would backfire and increase harassment by law enforcement.
And grassroots organizing and great vision, without a voice at the tables of power, is a stymied power. Paul Wellstone decided to run for office after years of frustrating fights around welfare, farm foreclosures, apartheid and veteran's benefits—so that the people of MN would have somebody in office on their side when they mobilized their communities around issues that impacted them.
Integrating the Triangle
When all three pieces of the triangle are working in concert, we build long-term movement and institutional power. Of course, at different times during any given cycle, certain actions and pieces of the triangle rise to greater importance. Last year elections took greater precedence. Our work on local, state and national races and ballot initiatives was an incredible opportunity for us to expand our base and engage people in conversations about their lives and what matters to them. These new relationships and conversations are the foundation from which we build our issues and policy campaigns moving forward. And in the next elections, new people we have brought in and leaders we have developed through our issue organizing will be instrumental in winning victories at the ballot box. That is how we build real, sustainable power.
Rishi Awatramani is Lead Organizer at Virginia New Majority (VNM). VNM is a member of the Right to the City Alliance. Rishi is on the US Social Forum National Planning Committee representing Leftist Lounge, has previously worked as a union and community organizer, and is a long-time activist with several organizations.
The dual objectives of 1) winning improvements in the lives of oppressed communities and 2) challenging US-led imperialism from within the US find their best chances for success if we are able to organize communities in not just effective and creative campaigns, but also if we're able to organize in large numbers. Social movements in this country, therefore, have the responsibility of 1) building fighting organizations made up of leaders and members that will struggle in solidarity with oppressed peoples of the world, and 2) organizing the majority of people in their communities, and ultimately in the country to support political change that progressively builds social justice.
Yet, most grassroots organizations struggle to organize more than a few hundred active members, leaving the objective of organizing large numbers of people unrealized. The labor movement, in theory is less interested in organizing politically advanced members and more in growing the sheer numbers of organized workers, continues to lose members instead.
Ground Shifting Beneath Our Feet
There are unprecedented opportunities in this moment to grow our mass-based organizations in the number of people involved, and in the scale of impact we have. For example, in Northern Virginia, where I organize, over 45% of the voting population are People of Color, and that number is growing. Many U.S. cities are majority or near-majority non-White. This is unprecedented in most big metropolitan areas. Additionally, Communities of Color, along with many White (in particular progressive White) people united around the issues of the Barack Obama campaign on a scale not seen since the Jesse Jackson campaigns of 1984 and 1988. Both of these trends show a budding new majority (based on both demographics and political beliefs) that fundamentally changes our organizing terrain.
These changes are mirrored by the mobilization of right wing consciousness amongst White communities that has cut across class. While many communities were already organized, the virulence of their racist, anti-socialist attacks have recently grown in response to our first Black president and his perceived progressivism.
New Tasks for a New Majority
To effectively transform these conditions into advantages for building social movements in this country, we must make it a priority to converge this growing majority of people into sustained political action through the electoral process. In this moment, electoral work provides us with the opportunity to engage people in a form of political action they are more likely to engage in than any other. We have to build new organizations (like Florida and Virginia New Majority) that can organize communities on a large scale through the electoral process to shape the future of their communities and the country in a way we haven't before.
The objectives of this work include involving thousands, if not millions of people in conscious political action, winning office for progressive candidates (including those that emerge directly from our base), training communities in direct accountability of elected officials we put into office, and sharpening our skills at running campaigns. The success of this work hinges on 1) using non-election time to organize communities to understand the electoral process as one step towards deeper forms of political change; and 2) involving the leadership from grassroots organizations in providing political leadership to the broad spectrum of people that will be mobilized through this work.
There are several challenges to this work: it requires massive resources; it's difficult to develop other campaigns because of the frequency and intensity of electoral cycles; voters are less likely to get involved when there are not exciting candidates; many people, including undocumented immigrants and felons can't vote; and it's possible to develop false hope in our ability to eradicate exploitation with our votes. We need creative solutions to these challenges.
We must not mistake the political power we might win through this process as analogous to the power people might win through deeper forms of political change. It is equally important that we recognize the potential to create real benefits for oppressed people in the US and beyond through this type of political work. And more than anything, we have to build new organizations for the new emerging majority in this country that can build towards deep, lasting social justice.
Charles Lenchner is co-founder of Organizing 2.0 and 20 year veteran of electoral and advocacy campaigning.
Systems built around candidates do a poor job of recruiting and training leaders. Most campaigns don't have the time or resources. Remember that much of what the Obama campaign did is not typical of electoral politics.
Electoral politics are rigged in favor of highly technical, top-down strategies that do not rely on mass participation. This holds true even when a relatively high proportion of money is spent on field work as opposed to advertising.
It's a consultant and media based culture in which regular citizens and activists are often held in contempt as 'amateurs.' In most races, incumbents win with the same combination of money, power players and local grasstops that brought them into office. 'Citizen empowerment' often translates into the rise and fall of very specific community groups and sectors, not an ethos in which people simply matter. It's a mindset that undermines small 'd' democracy.
That said, it's also true that challengers and folks transitioning into electoral politics from other arenas draw on the skills and tools of community organizing. So organizers with a grassroots bent can see some local electoral campaigns as helping to strengthen the progressive movement. The election in New York City of Brad Lander, Margaret Chin and Jumaane Williams are cases in point. But they are the exception.
It's also true that Presidential elections push a lot of money to specific GOTV efforts working with key demographics. The intersection of money, media attention and focus can be used to expand the circle of politically aware community members. I hope we see more career oriented grassroots organizers gaining experience in electoral politics to bring back some of the tools that work, especially online tools and databases.
Jessamyn Sabbag is a Bay Area native, currently based in Oakland. Jessamyn has been active in progressive social change work for the last decade. She cut her teeth in organizing through high impact anti-police brutality work in her hometown. She is currently Field Director of Oakland Rising, an up and coming alliance of social justice organizations employing electoral strategies to move the issues and agendas of low-income communities of color to the center of city government.
Over the last 8 months I have run two electoral field campaigns and a civic engagement program that has collectively impacted over 14,000 Oakland residents. As Field Director for Oakland Rising, I spend a lot of time thinking about the possible marriages between electoral and grassroots organizing. It's not an easy concept. 500 words is too short. But below I will examine three "marriages" that I have been trying to address in my work. And I'll show how Oakland Rising is intentionally working to develop integrated grassroots and electoral organizing to build the power we need to win, to move the issues and advance the agendas of low-income communities of color to the center of city government.
1) Culture Shifting: From Struggle to Winning
2) Quantity AND Quality: One Hand Washes the Other
3) Developing Leaders: Cross-Over Skills and Issues
1) Culture Shifting: From Struggle to Winning
Since coming to Oakland Rising as Field Director, I've learned a bunch of quotes to describe our tactics and strategy. One of my favorites is "We don't believe in struggle. We believe in winning." Oakland Rising is on the path towards developing a collaborative model that harnesses the scale we need for electoral power and the depth we need for grassroots progressive social change. But shifting the grassroots base and intermediary organizations from a model and history of struggle to a model and program based on winning at all costs takes time. We all agree theoretically that electoral organizing is different from grassroots organizing, including the realities of a short lead time for electoral planning, and a fast paced environment to achieve goals of significant scale. Over the last 9 months, I have had the opportunity to usher in culture shifts by developing models that integrate the science of electoral organizing with the equation to build grassroots power.
2) Quantity AND Quality
Oakland Rising is committed to developing the quantity we need to win at the ballot box and the quality of voters we need to hold elected officials accountable. In our latest campaign, our scale nearly doubled when we contacted over 12,000 Oakland voters about local campaigns like the development of a local transit hub. In our 2-4 minute electoral style conversations we were able to engage in political education and get community feedback (outside of the social justice "base") that helps redirect grassroots campaign framing and increase follow-up. And our base-building organizations are currently doing more in-depth outreach with voters who were IDed as "hot contacts."
3) Developing Leaders
Few things are more satisfying than a good win, right? Fortunately, electoral organizing offers a couple opportunities a year to get a good win (especially here in California where it seems like we have an election every other month!). While phoners and canvassers can do quick-hit issue education and identify the supporters we need to win on election day, moving that win into a community of leaders takes strategic grassroots organizing. Oakland Rising hires organizational volunteer leaders and community members to phone and knock as members of the electoral daily team. Volunteer leaders who work on our Daily Team developed or deepened a new skill set which is continually used to help with grassroots organization-specific campaigns.


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I am only selling this rifle because I have had it for 3-4 years and only shot 20- 40 Fed Gold Metal Match 168 grain rounds thru it to sight in the scope and it has been sitting in the safe ever since.
The rifle shoots .5 MOA groups with box ammo
This rifle is a Remington 700 built by Red Hawk Rifles. It is designed to be a light weight hunting rifle at 6.5 pounds. The barrel is approx 19 3/8 inches fluted and crowned.The barrel was cryo treated. The bolt is fluted and the bolt handle is drilled. The stock is a Bell & Carlson. The scope rail is Precision Armament 20 MOA.
This aution does not include the bipod, scope,scope rings, or the Bradley Cheek rest.Please call or text any questions or offers to 760-953-9067