text:
The present invention provides an aptamer-based calorimetric sensor system for determining the presence and optionally the concentration of an analyte in a sample. Methods of utilizing the sensor system and kits that include the sensor also are provided. The sensor utilizes a linker and oligonucleotide functionalized particles to form an aggregate, which disaggregates in response to the analyte.
1. A sensor system for detecting an analyte, comprising:
an aggregate comprising:
(a) a linker comprising an aptamer,
(b) first particles,
(c) a first oligonucleotide coupled to the first particles,
where the first oligonucleotide is hybridized to at least a portion of the aptamer,
the aptamer is capable of folding in response to the analyte, causing disaggregation of the aggregate, and the linker is an oligonucleotide.
2. The system of
3. The system of
4. The system of
5. The system of
6. The system of
7. The system of
8. The system of
9. A sensor system for detecting an analyte, comprising:
an aggregate, wherein the aggregate comprises a plurality of complexes,
wherein each complex comprises:
(I) a linker comprising an aptamer,
(II) first particles,
(II) a first oligonucleotide coupled to the first particles,
(III) second particles,
(IV) a second oligonucleotide coupled to the second particles,
where the first oligonucleotide is hybridized to at least a portion of the aptamer,
the second oligonucleotide is hybridized to at least a portion of the aptamer,
the aptamer is capable of folding in response to the analyte, causing disaggregation of the aggregate, and
the linker is an oligonucleotide.
10. The system of
11. The system of
12. The system of
13. The system of
14. The system of
15. The system of
16. The system of
This subject matter of this application may have been funded in part under the following research grants and contracts: National Science Foundation Contract Numbers CTS-0120978 and DMR-0117792. The U.S. Government may have rights in this invention.
The ability to determine the presence of an analyte in a sample is of significant benefit. For example, many metals and metal ions, such as lead, mercury, cadmium, chromium, and arsenic, pose significant health risks when present in drinking water supplies. To prevent the contamination of drinking and other water supplies, it is common to test industrial waste-streams before their release to the water treatment plant. Biological fluids, such as blood and those originating from body tissues, also may be tested for a variety of analytes to determine if the body has been exposed to harmful agents or if a disease state exists. For example, the need to detect trace amounts of anthrax in a variety of samples has recently emerged.
Colorimetric methods are commonly used for the detection of metals and ions in soil, water, waste-streams, biological samples, body fluids, and the like. In relation to instrument based methods of analysis, such as atomic absorption spectroscopy, calorimetric methods tend to be rapid and require little in the way of equipment or user sophistication. For example, colorimetric tests are available to aquarists that turn darker shades of pink when added to aqueous samples containing increasing concentrations of the nitrate (NO3−) ion. In this manner, colorimetric tests show that the analyte of interest, such as nitrate, is present in the sample and also may provide an indicator of the amount of analyte in the sample through the specific hue of color generated. While conventional colorimetric tests are extremely useful, they only exist for a limited set of analytes, and often cannot detect very small or trace amounts of the analyte.
As can be seen from the above description, there is an ongoing need for calorimetric sensor systems that can identify trace amounts of a broader scope of analytes and that increase the reliability of the analysis.
A sensor system for detecting an analyte includes a linker comprising an aptamer that folds in response to the analyte and second particles coupled to a second oligonucleotide that is complementary to at least a portion of the aptamer. The linker may include an extension where a first oligonucleotide coupled to first particles is complementary to at least a portion of the extension.
A method of detecting an analyte includes combining an aggregate with a sample to detect a color change responsive to the analyte. The aggregate may include a linker and second particles. The aggregate also may include first particles and the linker may include an extension.
A kit for detecting an analyte includes a first container containing a system for forming aggregates that includes second particles and a linker including an aptamer, which folds in response to the analyte. The second particles are coupled to second oligonucleotides that are complementary to at least a portion of the aptamer.
A method for determining the sensitivity and selectivity of an aptamer to an analyte includes combining an aggregate with the analyte, detecting a color change responsive to the analyte, and determining if the DNA strand folded to provide the color change. The aggregate includes second particles and a linker including a DNA strand. The aggregate also may include first particles. The linker may include an extension.
In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided.
The term "sample" is defined as a composition that will be subjected to analysis that is suspected of containing the analyte of interest. Typically, a sample for analysis is in a liquid form, and preferably the sample is an aqueous mixture. A sample may be from any source, such as an industrial sample from a waste-stream or a biological sample, such as blood, urine, or saliva. A sample may be a derivative of an industrial or biological sample, such as an extract, a dilution, a filtrate, or a reconstituted precipitate.
The term "analyte" is defined as one or more substance potentially present in the sample. The analysis determines the presence, quantity, or concentration of the analyte present in the sample.
The term "calorimetric" is defined as an analysis where the reagent or reagents constituting the sensor system produce a color change in the presence or absence of an analyte.
The term "light-up" refers to a colorimetric sensor system that undergoes a desired color change in response to an analyte present in a sample.
The term "light-down" refers to a colorimetric sensor system that does not undergo a color change when an analyte is present in a sample, but does undergo a desired color change in the absence of the analyte.
The term "sensitivity" refers to the smallest increase in an analyte concentration that is detectable by the sensor system (resolution) or to the lowest concentration limit at which a sensor system can differentiate a signal responsive to the analyte from a background signal (detection limit). Thus, the more sensitive a sensor system is to an analyte, the better the system is at detecting lower concentrations of the analyte.
The term "selectivity" refers to the ability of the sensor system to detect a desired analyte in the presence of other species.
The term "hybridization" refers to the ability of a first polynucleotide to form at least one hydrogen bond with at least one second nucleotide under low stringency conditions.
The term "aptamer" refers to a strand of nucleic acids that undergoes a conformational change in response to an analyte.
The term "conformational change" refers to the process by which an aptamer adopts a tertiary structure from another state. For simplicity, the term "fold" may be substituted for conformational change.
The invention can be better understood with reference to the following drawings and description. The components in the figures are not necessarily to scale and are not intended to accurately represent molecules or their interactions, emphasis instead being placed upon illustrating the principles of the invention.
Aptamers may be easier to isolate than nucleic acid based catalysts. The simpler structure of aptamers in relation to nucleic acid enzymes also may allow for the design of analyte sensor systems for which nucleic acid enzymes are not available. The present invention makes use of the discovery that by selecting the hybridization strength between the folded and unfolded conformations of an aptamer and an oligonucleotide functionalized particle, the particle may be released in response to an analyte. In this manner, a light-up calorimetric sensor is provided that undergoes a desired color change in response to a selected analyte at room temperature, thus overcoming a disadvantage of the sensor system disclosed in U.S. Ser. No. 10/144,679.
In one aspect, the analyte 105 may be any ion that causes an aptamer 124 to fold. In another aspect, the analyte 105 may be any metal ion that causes an aptamer 124 to fold. Preferable monovalent ions having a +1 formal oxidation state (I) include NH4 +, K(I), Li(I), Tl(I), and Ag(I). Preferable divalent metal ions having a +2 formal oxidation state (II) include Mg(II), Ca(II), Mn(II), Co(II), Ni(II), Zn(II), Cd(II), Cu(II), Pb(II), Hg(II), Pt(II), Ra(II), Sr(II), Ni(II), and Ba(II). Preferable trivalent and higher metal ions having +3 (III), +4 (IV), +5 (V), or +6 (VI) formal oxidation states include Co(III), Cr(III), Ce(IV), As(V), U(VI), Cr(VI), and lanthanide ions. More preferred analyte ions include monovalent metal ions and metal ions that are toxic to living organisms, such as Ag(I), Pb(II), Hg(II), U(VI), and Cr(VI).
In another aspect, the analyte 105 may be any biomolecule that causes the aptamer 124 to fold. Preferable biomolecules include large biomolecules, such as proteins (e.g. proteins related to HIV, hCG-hormone, insulin), antibodies, growth factors, enzymes, virus (e.g. HIV, small pox), viral derived components (e.g. HIV-derived molecules), bacteria (e.g. anthrax), bacteria derived molecules and components (e.g. anthrax derived molecules), or cells. Preferable biomolecules also may include small biomolecules, such as amino acids (e.g. arginine), nucleotides (e.g. ATP, GTP), neurotransmitters (e.g. dopamine), cofactors (e.g. biotin), peptides, or amino-glycosides.
In another aspect, the analyte 105 may be any organic molecule that causes the aptamer 124 to fold. Preferable organic molecules include drugs, such as antibiotics and theophylline, or controlled substances, such as cocaine, dyes, oligosaccharides, polysaccharides, glucose, nitrogen fertilizers, pesticides, dioxins, phenols, 2,4-dichlorophenoxyacetic acid, nerve gases, trinitrotoluene (TNT), or dinitrotoluene (DNT).
Once the analyte 105 is selected, the one or more aptamer 124 is selected that folds in response to the analyte 105. The aptamer selection 120 may be performed by in vitro selection, directed evolution, or other method known to those of ordinary skill in the art. The aptamer selection 120 may provide one or more aptamers that demonstrate enhanced folding in the presence of the selected analyte 105 (thereby providing sensor sensitivity). The selection 120 also may exclude aptamers that fold in the presence of selected analytes, but that do not fold in the presence of non-selected analytes and/or other species present in the sample 102 (thereby providing sensor selectivity).
For example, an aptamer may be selected that specifically binds K(I), while not significantly binding Na(I), Li(I), Cs(I), Rb(I), or other competing metal ions. In one aspect, this may be achieved by isolating aptamers that bind K(I), then removing any aptamers that bind Na(I), Li(I), Cs(I), or Rb(I). In another aspect, aptamers that bind Na(I), Li(I), Cs(I), or Rb(I) are first discarded and then those that bind K(I) are isolated. In this manner, the selectivity of the aptamer may be increased.
The aptamer 124 includes a nucleic acid strand that folds in the presence of the analyte 105. In one aspect, the folding may be considered the conversion of a primary or duplex structure to a tertiary structure. The base sequence of the aptamer may be designed so that the aptamer may undergo at least partial hybridization with at least one oligonucleotide functionalized particle. In this aspect, at least a portion of the base sequence of the aptamer 124 may be complementary to at least one oligonucleotide of the oligonucleotide functionalized particle.
The aptamer 124 may be formed from deoxyribonucleotides, which may be natural, unnatural, or modified nucleic acids. Peptide nucleic acids (PNAs), which include a polyamide backbone and nucleoside bases (available from Biosearch, Inc., Bedford, Mass., for example), also may be useful.
Table I below lists analytes, the aptamer or aptamers that bind with and fold in response to that analyte, and the reference or references where the sequence of each aptamer is described. The analyte binding region of these, and other, aptamers may be adapted for use in a linker 128. For example, the non-analyte binding region of the cocaine aptamer, given as SEQ ID NO. 10 in Table I below, may be modified to provide the aptamer GGGAGACAAGGATAAATCCTTCAATGAAGTGGGTCTCCC (SEQ ID NO. 56) and included in the linker 128.
After selecting an appropriate aptamer or aptamers in 120, a linker 128 is formed that includes the aptamer 124. In one aspect, the aptamer 124 may serve directly as the linker 128. In another aspect, the linker 128 may be formed by joining the aptamer 124 with one or more extensions 126.
The extension 126 may be any nucleic acid sequence that may be joined with the aptamer 124, that may undergo at least partial hybridization with one or more oligonucleotide functionalized particles, and that is compatible with the analysis 100. In this aspect, at least a portion of the base sequence of the extension 126 may be complementary to at least one oligonucleotide of one or more oligonucleotide functionalized particle. In one aspect, solid phase synthesis may be used to join the aptamer 124 with the extension 126 to form the linker 128. In another aspect, after the aptamer 124 portion of the linker 128 is synthesized, the synthesis is continued to form the extension 126. Similarly, the linker 128 may be extended with the aptamer 124 sequence.
Preferably, the extension 126 includes from 1 to 100 bases. In one aspect, at least 50, 70, or 90% of the bases present in the extension 126 are capable of hybridizing with a complementary portion of a first oligonucleotide functionalized particle, such as the TGAGTAGACACT-5′ (SEQ ID NO. 43) portion of particle 336 in
After selecting or synthesizing the linker 128, an aggregate 132 may be formed in 130. The aggregate 132 includes the linker 128 and oligonucleotide functionalized particles 136. Considering the physical size of its components, the aggregate 132 may be quite large.
The linker 128 hybridizes with the oligonucleotide functionalized particles 136 and includes the aptamer 124 and may include the extension 126. For example, if first and second oligonucleotide functionalized particles have base sequences of 3′-AAAAAAAAAAAATGAGTAGACACT (SEQ ID NO. 44) and 5′-CCCAGGTTCTCT (SEQ ID NO. 45), respectively, an appropriate sequence for the linker 128 that includes the aptamer 124 that folds in the presence of an adenosine analyte and the extension 126 may be 5′-ACTCATCTGTGAAGAGAACCTGGGGGAGTATTGCGGAGGAAGGT (SEQ ID NO. 46).
For the adenosine analyte, the extension 126 portion of the linker 128 is the ACTCATCTGTGAAGAGA (SEQ ID NO. 47) portion of the sequence, which allows the extension 126 to hybridize with twelve bases of the first functionalized particle and five bases of the second functionalized particle. Similarly, the aptamer 124 portion of the linker 128 is the ACCTGGGGGAGTATTGCGGAGGAAGGT (SEQ ID NO. 18) portion of the sequence, which allows the ACCTGGG (SEQ ID NO: 48) portion of the aptamer 124 to hybridize with the TGGACCC (SEQ ID NO. 49) portion of the second functionalized particle.
Because the particles 136 demonstrate distance-dependent optical properties, the particles are one color when closely held in the aggregate 132 and undergo a color change as the distance between the particles increases. For example, when the particles 136 are gold nanoparticles, the aggregate 132 displays a blue color in aqueous solution that turns red as disaggregation proceeds.
Disaggregation occurs when the aptamer 124 portion of the linker 128 binds with and folds in response to the analyte 105. When the aptamer 124 folds, a portion of the hybridization with the second oligonucleotide functionalized particles is lost. This hybridization loss allows the second oligonucleotide functionalized particles to separate from the aggregate 132. Thus, as the particles 136 diffuse away from the aggregate 132, the solution changes from blue to red.
The particles 136 may be any species that demonstrate distance-dependent optical properties and are compatible with the operation of the sensor system. Suitable particles may include metals, such as gold, silver, copper, and platinum; semiconductors, such as CdSe, CdS, and CdS or CdSe coated with ZnS; and magnetic colloidal materials, such as those described in Josephson, Lee, et al., Angewandte Chemie, International Edition (2001), 40(17), 3204-3206. Specific useful particles may include ZnS, ZnO, TiO2 , Agl, AgBr, HgI2, PbS, PbSe, ZnTe, CdTe, In2S3, In2Se3, Cd3P2 , Cd3AS2, InAs, and GaAs.
In a preferred aspect, the particles are gold (Au) nanoparticles and have an average diameter from 5 to 70 nanometers (nm) or from 10 to 50 nm. In a more preferred aspect, gold nanoparticles having an average diameter of from 10 to 15 nm are functionalized to the oligonucleotides.
For a more detailed treatment of how to prepare oligonucleotide functionalized gold particles, See U.S. Pat. No. 6,361,944; Mirkin, et al., Nature (London) 1996, 382, 607-609; Storhoff, et al., J. Am. Chem. Soc. 1998, 20, 1959-1064; and Storhoff, et al., Chem. Rev. (Washington, D.C.) 1999, 99, 1849-1862. While gold nanoparticles are presently preferred, other species that undergo a distance-dependent color change, such as inorganic crystals, quantum dots, and the like also may be attached to oligonucleotides.
In 140 the aggregate 132 may be combined with the sample 102. In 150 the sample 102 is monitored for a color change. If a color change does not occur, then the analyte 105 is not present in the sample 102. If a color change does occur in 160, the analyte 105 is present in the sample 102. The color change signifies that the analyte 105 caused the aptamer 124 to fold, thus allowing the particles 136 to diffuse away from the aggregate 132 and into the solution of the sample 102. Thus, the analysis 100 provides a "light-up" sensor system because a color change occurs in the presence of the analyte 105.
The rate at which a substantially complete color change occurs in response to the analyte 105 may be considered the response time of the sensor system. In one aspect, the color change may be considered substantially complete when the absorption peak in the visible region increases by 20%. In another aspect, the color change may be considered substantially complete when the extinction coefficient at 522 nm over 700 nm increases by 100% for gold particles. Preferable response times for the sensor system are from 1 second to 60 minutes or from 2 seconds to 10 minutes. More preferable response times for the sensor system are from 5 seconds to 2 minutes or from 8 to 12 seconds. Preferable temperature ranges for operation of the sensor system are from 0� to 60� or from 15� to 40� C. More preferable ranges for operation of the sensor system are from 23� to 37� or from 25� to 30� C. In another aspect, when the analysis 100 is conducted from 23� to 37� C., a preferable response time may be less than 2 minutes or from 1 to 12 seconds.
The degree the color changes in response to the analyte 105 may be quantified by colorimetric quantification methods known to those of ordinary skill in the art in 170. Various color comparator wheels, such as those available from Hach Colo., Loveland, Colo. or LaMotte Colo., Chestertown, Md. may be adapted for use with the present invention. Standards containing known amounts of the selected analyte may be analyzed in addition to the sample to increase the accuracy of the comparison. If higher precision is desired, various types of spectrophotometers may be used to plot a Beer's curve in the desired concentration range. The color of the sample may then be compared with the curve and the concentration of the analyte present in the sample determined. Suitable spectrophotometers include the Hewlett-Packard 8453 and the Bausch & Lomb Spec-20.
In yet another aspect, the method 100 may be modified to determine the sensitivity and selectivity of an aptamer to the analyte 105. In this aspect, one or more DNA strand suspected of being an aptamer responsive to the analyte 105 is selected in 120. The DNA strand may or may not be modified with the extension 126 in 125. In 130, an aggregate is formed from the DNA strands and appropriate particles. In 140, the aggregates are combined with the analyte 105. If the aggregate undergoes a color change, then the DNA strand is an appropriate aptamer sequence for the target 105. In this manner, multiple aptamers selected in 120 may be tested for use in a colorimetric sensor system.
The aptamer 224 includes C and D portions. In one aspect, the hybridization stability of the combined B portion of the extension 226 and the C portion of the aptamer 224 with the second particle 237 may be less than that for A and the first particle 236. In a preferred aspect, the melting temperature of this C+B/second particle hybridization is higher than the temperature at which the sensor system is to be used. In another preferred aspect, the melting temperature of a B portion hybridized to the oligonucleotide sequence of the second particle 237 is less than the temperature at which the sensor system is to be used. In another preferred aspect, the stability of a C+D+ analyte complex should be greater than the hybridization stability of C with the second particle 237. In another aspect, the sequences of B and C are as short as is compatible with the operation of the sensor system.
While one base sequence for the linker and the particles are shown, the bases may be changed on the opposing strands to maintain the pairings. For example, any cytosine in the A or B portions of the linker 228 may be changed to thymine, as long as the paired base of the particle oligonucleotide is changed from guanine to adenine.
In the presence of the adenosine analyte 305, the aptamer portion 324 of the linker 328 folds, thus eliminating at least a portion of the hybridization between the aptamer portion 324 of the linker 328 and the 5′-AdeAu particle 337. This loss of hybridization between the aptamer 324 and the 5′-AdeAu particle 337 releases the 5′-AdeAu particle to the solution.
As the 5′-AdeAu particles 337 are released, the blue aggregate 332 begins to disaggregate to form partial aggregate 390. This partial disaggregation adds red color to the blue solution as the particles 337 diffuse away from the aggregate 332, thus giving a purple solution. If enough of the adenosine analyte 305 is present in the sample, the reaction will continue until the aggregate 332 is completely disaggregated, to give 395. Complete disaggregation results in a red solution due to the greater distance between the particles 336, 337.
The alignment of the particles 336, 337 (tail-to-tail, head-to-tail, or head-to-head) with respect to each other may influence how tightly the aggregate units 331 bind.
At present, the tail-to-tail hybridization arrangement of
While not shown, the methodology of
The ionic strength of the sample may influence how tightly the moieties that form the aggregate bind together. Higher salt concentrations favor aggregation, thus slowing sensor response, while lower salt concentrations may lack the ionic strength necessary to maintain the aggregates. In one aspect, the sample may include or be modified with a reagent to include a monovalent metal ion concentration of 30 mM and greater. The ionic strength of the sample may be modified with Na+ ions, for example. In a preferred aspect, the monovalent metal ion concentration of the sample, which contains the aggregate, is from 150 to 400 mM. At present, especially preferred monovalent metal ion concentrations are about 300 mM for adenosine and potassium analytes and about 150 mM for cocaine as an analyte. pH also may influence the aggregate binding, possibly attributable to the protonation of the polynucleotide base pairs at lower pH. In one aspect, a pH from 5 to 9 is preferred, with an approximately neutral pH being more preferred.
Thus, the performance of the sensor may be improved by adjusting the ionic strength and pH of the sample prior to combining it with the aggregate. Depending on the sample, it may be preferable to add the sample or analyte to a solution containing the aggregate (where the ionic strength and pH may be controlled) or the reverse.
The sensor system, including the aptamer, extension, and oligonucleotide functionalized particles may be provided in the form of a kit. In one aspect, the kit includes the aptamer and the extension joined to form the linker. In yet another aspect, the kit includes the extension, but excludes the aptamer, which is then provided by the user or provided separately. In this aspect, the kit also may include the reagents required to link the supplied extension with an aptamer. In this aspect, the kit also may be used to determine the specificity and/or selectivity of various aptamers to a selected analyte. Thus, the kit may be used to select an appropriate aptamer in addition to detecting the analyte. In yet another aspect, the kit includes an exterior package that encloses a linker and oligonucleotide functionalized particles.
One or more of these kit components may be separated into individual containers, or they may be provided in their aggregated state. If separated, the aggregate may be formed before introducing the sample. Additional buffers and/or pH modifiers may be provided in the kit to adjust the ionic strength and/or pH of the sample.
The containers may take the form of bottles, tubs, sachets, envelopes, tubes, ampoules, and the like, which may be formed in part or in whole from plastic, glass, paper, foil, MYLAR�, wax, and the like. The containers may be equipped with fully or partially detachable lids that may initially be part of the containers or may be affixed to the containers by mechanical, adhesive, or other means. The containers also may be equipped with stoppers, allowing access to the contents by syringe needle. In one aspect, the exterior package may be made of paper or plastic, while the containers are glass ampoules.
The exterior package may include instructions regarding the use of the components. Color comparators; standard analyte solutions, such as a 10 μm solution of the analyte; and visualization aids, such as thin layer chromatography (TLC) plates, test tubes, and cuvettes, also may be included. Containers having two or more compartments separated by a membrane that may be removed to allow mixing may be included. The exterior package also may include filters and dilution reagents that allow preparation of the sample for analysis.
In another aspect, in addition to the sensor system of the present invention, the kit also may include multiple sensor systems to further increase the reliability of analyte determination and reduce the probability of user error. In one aspect, multiple light-up sensor systems in accord with the present invention may be included. In another aspect, a "light-down" sensor system may be included with the light-up sensor system of the present invention. Suitable light-down sensors for inclusion in the presently claimed kit may rely on DNAzyme/Substrate/particle aggregates that are not formed in the presence of the selected analyte. A more detailed description of suitable light-down sensor systems for inclusion in the presently claimed kit may be found, for example, in U.S. patent application Ser. No. 10/756,825, filed Jan. 13, 2004, entitled "Biosensors Based on Directed Assembly of Particles," which is hereby incorporated by reference.
The preceding description is not intended to limit the scope of the invention to the described embodiments, but rather to enable a person of ordinary skill in the art to make and use the invention. Similarly, the examples below are not to be construed as limiting the scope of the appended claims or their equivalents, and are provided solely for illustration. It is to be understood that numerous variations can be made to the procedures below, which lie within the scope of the appended claims and their equivalents.
All DNA samples were purchased from Integrated DNA Technology Inc., Coralville, Iowa. The DNA samples that formed the extensions were purified by gel electrophoresis, while the thiol-modified DNA samples for forming the oligonucleotide functionalized particles were purified by standard desalting. Adenosine, cytidine, uridine, guanosine and cocaine were purchased from Aldrich. Gold nanoparticles having an average diameter of 13 nm were prepared and functionalized with 12-mer thiol-modified DNA following literature procedures, such as those disclosed in Storhoff, J., et al., "One-pot calorimetric differentiation of polynucleotides with single base imperfections using gold particle probes," JACS 120: 1959-1964 (1998), for example. The average diameter of the gold nanoparticles was verified by transmission electronic microscope (JEOL 2010).
Preparation of a Colorimetric Adenosine Sensor
Five-hundred microliters of 5′-AdeAu (extinction at 522 nm equals 2.2) and 500 μL of 3′-A12AdeAu (SEQ ID NO: 44) (extinction at 522 nm equals 2.2) were mixed in the presence of 300 mM NaCl, 25 mM Tris acetate buffer, pH 8.2, and 100 nM of the adenosine aptamer/extension (Adenosine Linker). The sample was warmed to 65� C. for one minute and then allowed to cool slowly to 4� C. The nanoparticles changed color from red to dark purple during this process. The sample was centrifuged and the precipitates were collected and then dispersed in 2000 μL of the same buffer (300 mM NaCl, 25 mM Tris acetate, pH 8.2). The suspension was used for detection of adenosine.
Colorimetric Detection of Adenosine
One-hundred microliters of the sensor suspension from Example 1 was added to a small volume of concentrated adenosine solution. For example, 2 μL of 50 mM adenosine was added to give a final concentration of 1 mM. The color change was monitored with a UV-vis spectrometer or by the naked eye.
Monitoring the Performance of the Adenosine Sensor
The color change of the sample from Example 2 was monitored by UV-vis extinction spectroscopy.
Selectivity and Sensitivity of the Adenosine Sensor
To a quarts UV-vis spectrophotometer cell (Hellma, Germany) was added 100 μL of the adenosine sensor system prepared in Example 1. A small volume (1-5 μL) of solutions including adenosine, uridine, cytidine or guanosine was added to the spectrophotometer cell to bring the analyte concentration to the desired level in the cell.
The dispersion kinetics for each cell was monitored as a function of time using a Hewlett-Packard 8453 spectrophotometer.
In addition to the instrumental method of
Preparation of a Colorimetric Potassium Ion Sensor
Colorimetric Detection of Potassium
Metal ion solutions of Li+, Na+, K+, Rb+, or Cs+ ions were made by dissolving LiCl, NaCl, KCl, RbCl or CsCl salt, respectively, in deionized water to obtain an ion concentration of 3 M. From these metal ion stock solutions were prepared solutions containing 25 mM of Tris acetate buffer, pH 8.2, and 300 mM of Li+, Na+, K+, Rb+, or Cs+ ions. To each of these five solutions was added 1 μL of the K(I) sensor system from Example 5 for each 99 μL of the metal ion containing solution. Therefore, each solution contained ˜300 mM of Li+, Na+, K+, Rb+, or Cs+ metal ions and an additional 3 mM of Na+ ions as background. Each sample was heated to 65� C. and then cooled slowly to 4� C. in 1 hour. The color change was monitored with a UV-vis spectrometer or by the naked eye.
Preparation of a Colorimetric Cocaine Sensor
Colorimetric Detection of Cocaine
One-hundred microliters of the above prepared cocaine sensor suspension were combined with a small volume of concentrated cocaine solution. For example, 1 μL of 100 mM cocaine was added to the suspension to give a final concentration of 1 mM. The color change was monitored with a UV-vis spectrometer or by the naked eye.
Selectivity and Sensitivity of the Cocaine Sensor
To a quarts UV-vis spectrophotometer cell (Hellma, Germany) was added 100 μL of the cocaine sensor system prepared in Example 7. A small volume (0.5-2 μL) of solutions including cocaine, adenosine, or sucrose was added to the spectrophotometer cell to bring the analyte concentration to the desired level in the cell.
The dispersion kinetics for each cell was monitored as a function of time using a Hewlett-Packard 8453 spectrophotometer.
In addition to the instrumental method of
As any person of ordinary skill in the art will recognize from the provided description, figures, and examples, that modifications and changes can be made to the preferred embodiments of the invention without departing from the scope of the invention defined by the following claims and their equivalents.
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A few weeks ago, we outlined the Top 5 Non-Compete Issues to Watch For In 2011. Among our top five, we noted that non-compete legislation stalled in the Massachusetts Legislature in 2010, and we forecasted that new and modified legislation would be introduced in 2011. Sure enough, a new bill was recently introduced. It contains numerous changes from the prior legislation, and it addresses issues ranging from consideration to garden leave to inevitable disclosure. An excellent summary of the proposed legislation along with a description of the changes from last year's bill is available on Russell Beck's blog at FairCompetitionLaw.com.
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CentOS Admin Mysteriously Missing
An open letter on the CentOS distribution website gives the impression that their main administrator has mysteriously disappeared.
Lance Davis is mainly in charge of the Linux distro CentOS, a re-build from Red Hat Enterprise Linux. In an open letter aimed at him, seven developers are appealing for him to get in touch. Efforts to contact Davis by phone have been of no avail. Davis holding the domain plus sole 'Founders' rights in the IRC channels makes the situation particularly difficult for the developers. Davis obviously is also in charge of donation money. In the letter: "You have long promised a statement of CentOS project funds, to this date this has not appeared."
Developers at CentOS are worried that Davis' AWOL will endanger the whole project: "Please do not kill CentOS through your fear of shared management of the project."
CentOS AccountingHas there been an accounting and disclosure of funds collected and disbursed? Money collected for non-profits must be accounted for on IRS form 990. Has one been created? What is the non-profit name collecting money for CentOS?
I gave money to CentOS and want an accounting.
CentOS Admin Mysteriously MissingUmm, no.
He is not "mysteriously missing" and he is *not* the main administrator of the project. He just has his fingers on some central things - the project has survived for the last year without him surfacing.
Actually he hasn't done any work on CentOS for quite some time.
The project itself is well alive and kicking.
Issue 175/2015
Buy this issue as a PDF
Code.org and College Board Team Reach Out for Talented High School Coders
New partnership will bring more and better CS training to US schools
A Peek Inside TeslaCrypt Ransomware
Criminals offer online help over Tor network
Mumblehard Still Spams
Sophisticated malware is still present on Joomla and WordPress sites around the world.
Launchpad Gets Git
Future versions of Ubuntu's code service will support the popular Git version control system used with Linux and other open source projects.
AMD Posts Code for Linux GPU Driver
New release marks the arrival of AMD's unified driver strategy.
30 Percent of New Server Equipment Goes to Cloud Infrastructure
A new study by IDC charts big changes in the big hardware market.
Windows Might Go Open Source
Azure CTO says Redmond has already considered the unthinkable.
Linux Mint Will Stay with Ubuntu
Lead developer quells rumors that the Debian version is slated for center stage.
Microsoft Frees Code for .NET Build Engine
MSBuild is now just another GitHub project as Redmond continues its path to the light.
Researchers Demonstrate Hack by Heat
Malware could pass data and commands between disconnected computers without leaving a trace on the network.


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Emily Wells is one woman with the force of a band — and her sound has evolved as much as her setup. Starting with a loop pedal and a violin, she's incorporated additional instrumentation to add depth to her hip-hop-influenced style, which is one part Biggie and two parts lullaby. Recently she returned to Los Angeles and joined Morning Becomes Eclectic to play a version of the excellent track "Mama's Gonna Give You Love."
Watch KCRW's entire session with Emily Wells at KCRW.com.
Source: NPR
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Rep. Buerkle: VA, DoD must 'bridge gap' in wounded warrior care
Friday - 5/20/2011, 5:03pm EDT
By Jolie Lee
Federal News Radio
It's been four years since the Departments of Defense and Veterans Affairs launched the Federal Recovery Coordinator Program, designed to expedite the process for wounded vets to get the care and services they need.
But one lawmaker says the two agencies are operating too independently of each other to make the program successful.
In opening remarks last week at the House Veteran's Affairs Subcommittee on Health, chairman Rep. Anne Marie Buerkle (D-N.Y.) said, "It appears that rather than having the joint program envisioned by the Commission to advocate on behalf of wounded warriors and ensure comprehensive and seamless rehabilitation, recovery, and transition, we have two separate programs - a VA program that utilizes Federal Recovery Coordinators and a DoD program that utilizes Recovery Care Coordinators."
The intent of the program was to "bridge that gap," Buerkle said in an interview with In Depth.
She said she was "dismayed" that the two agencies are "just talking about things and it seems like nothing's being done."
"Time is of the essence for these men and women coming out of the service and making that transition," she added.
A March 2011 report by the Government Accountability Office found the program still faces "significant challenges." One of those challenges is neither DoD nor VA's health IT systems classify servicemembers and vets as "severely wounded, ill, and injured," the report said. Therefore, it's unclear to determine who is in need of the FRCP referrals.
Buerkle said her main concern with the GAO report was that recommendations were directed at VA alone, and not both VA and DoD.
"The intent in this law was to have a joint program so the transition is seamless for our military," she said.
The hearing last week included VA and DoD outlining specific steps they would take to improve the program.
"That's the beauty of these hearings. It creates a record. Here's what was promised and now we need to have it delivered on," Buerkle said.
Faster VA-DoD disability evaluations still far off


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This Date In Baseball
1983 - Philadelphia's Steve Carlton struck out Lonnie Smith of the St. Louis Cardinals in the third inning for career strikeout No. 3,522, overtaking Nolan Ryan as the career strikeout king. St. Louis, however, beat the Phillies, 2-1.
1989 - Ernie Whitt had three hits and drove in three runs as the Toronto Blue Jays beat the Milwaukee Brewers 4-2 in the first game in major league history played indoors and outdoors on the same day. With the threat of rain, the SkyDome's $100 million retractable roof was closed.
2006 - Pitcher Jason Grimsley was released by the Arizona Diamondbacks, a day after his home was searched by federal agents following his admission he used human growth hormone, steroids and amphetamines.
2006 - Colorado tied a major league record with five sacrifice flies in a 16-9 win over Pittsburgh. Seattle set the mark against Oakland on Aug. 7, 1988.
2007 - Curt Schilling came within one out of his first career no-hitter, losing his bid when Shannon Stewart lined a clean single to right field in Boston's 1-0 victory over the Oakland Athletics. Schilling retired Mark Ellis on a foul pop to end the third one-hitter of his career.
2007 - Joe Torre became the 10th manager in major league history to get 2,000 victories as the New York Yankees defeated the Chicago White Sox 10-3.
2008 - Johnny Damon went 6-for-6 with a game-winning single in the bottom of the ninth inning, rallying the New York Yankees to a wild 12-11 victory over Kansas City.
2009 - Mark Reynolds hit a three-run homer off infielder Josh Wilson with two outs in the 18th inning, sending Arizona to a 9-6 win over San Diego. The Padres were held hitless through nine extra innings by four relievers. Their only baserunners in extras came on three walks. Jon Rauch pitched one inning, Esmerling Vasquez two, Clay Zavada 2 2-3 and Leo Rosales 3 1-3.
Today's birthday: Mark Lowe 28.
June 8
1914 - New York's Iron Joe McGinnity posted his 14th straight win beating Pittsburgh 2-0. With the win moved the Giants into first place over Chicago.
1927 - New York's Tony Lazzeri hit three homers in the Yankees 12-11 11-inning win over the Chicago White Sox. Lazzeri's first two homers come off Red Faber and his third was a two-run line drive off George Connally to tie game in the ninth inning. The Yanks were behind 11-6 going into the last inning. New York would win it in the 11th after Cedric Durst tripled Lazzeri was intentionally walked and Ray Morehart singled.
1933 - Philadelphia's Jimmie Foxx homered in his first three at bats all off Lefty Gomez as the A's beat the New York Yankees 14-10. Foxx had homered his last time up the previous day to tie a major league record of hitting four consecutive home runs. Bobby Lowe did it in 1894.
1940 - Harry Craft of Cincinnati connected for a home run, a triple, a double and two singles in seven at-bats to lead a 27-hit attack as the Reds pounded the Dodgers 23-2 at Brooklyn.


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Wikileaks Topics
Democracy Now! stories, posts and pages that relate to Wikileaks
- As Ecuador prepared to announce its decision on granting asylum to WikiLeaks founder Julian Assange, Britain threatened to raid the Ecuadorean embassy in London where Assange has taken refuge for the past two months. Britain told Ecuador that giving Julian Assange asylum would not change a thing and that it might still revoke the diplomatic status of Quito's embassy in London to allow the extradition of the WikiLeaks founder to Sweden to...August 16, 2012 | Story
- Michael Ratner, a member of WikiLeaks founder Julian Assange's legal team, reacts to the breaking news that Ecuador has approved Assange's request for political asylum two months after Assange took refuge in its London embassy. Britain says it still plans to extradite Assange to Sweden, where he faces questioning for alleged sexual misconduct. "The British ought to just back off, and the U.S. ought to just back off,"...August 16, 2012 | Story
- If Julian Assange is granted asylum in Ecuador, he will become a resident of Latin America, where the trove of classified U.S. State Department cables he strategically disseminated through WikiLeaks has generated hundreds of headlines, from Mexico to Chile. A year after thousands of cables on Latin America were first released, the revelations have had different results — in two countries it led to the forced departure of the U.S....August 03, 2012 | Story
Glenn Greenwald: As WikiLeaks Reveals Syria Files, Assange Remains in Ecuador Embassy Seeking AsylumWikiLeaks announced today it has begun publishing the Syria Files – more than two million emails from Syrian political figures, ministries and associated companies, dating from August 2006 to March 2012. WikiLeaks founder Julian Assange could not attend today's press conference announcing the release of the data trove because he is still inside the Ecuadorean embassy in London in an attempt to avoid extradition to Sweden. According to...July 05, 2012 | Story
- WikiLeaks founder Julian Assange has taken refuge in Ecuador's embassy in London and asked for asylum. Assange made the move Tuesday in a last-ditch bid to avoid extradition to Sweden over sex crime accusations. Earlier today, police in London announced Assange is now subject to arrest because his decision to spend the night at the Ecuadorian embassy violated the conditions of his bail. Assange is seeking asylum because he fears...June 20, 2012 | Story
- For just the third time since he was arrested over two years ago, alleged Army whistleblower Bradley Manning was seen by the public this week at a pretrial hearing in a military court at Fort Meade, Maryland. The 24-year-old private is accused of leaking hundreds of thousands of documents to the whistleblowing website WikiLeaks, including secret files on the Iraq and Afghan wars. Manning's attorneys are seeking the dismissal of 10 of the...June 08, 2012 | Story
"Private": Bradley Manning's Story, From Difficult Childhood to Alleged Whistleblower on World StageThe new book, "Private: Bradley Manning, WikiLeaks, and the Biggest Exposure of Official Secrets in American History," tracks Manning's trajectory from growing up as a gay teen in small-town Oklahoma to joining the U.S. Army, where he found success as an intelligence analyst before being charged with the largest U.S. intelligence breach on record. We speak with the book's author, Denver Nicks. "In many ways Bradley...June 08, 2012 | Story
- By Amy Goodman with Denis MoynihanMay 31, 2012 | Columns & Articles
The cases of Pvt. Bradley Manning, Julian Assange and former Chilean dictator Augusto Pinochet remind us that all too often whistle-blowers suffer, while war criminals walk.
- Britain's Supreme Court has upheld the extradition of WikiLeaks founder Julian Assange to Sweden to face questioning over alleged sex crimes. Assange's lawyers had argued that the Swedish public prosecutor did not have the legal authority to issue the arrest warrant, but the British justices disagreed in a 5-to-2 decision. Assange's attorneys will have 14 days to file a new appeal. We get reaction from Salon.com blogger and...May 30, 2012 | Story
- Helena Kennedy, a member of Julian Assange's legal team and a prominent British civil liberties attorney, joins us from London to discuss the court ruling upholding Assange's extradition to Sweden. "The idea of a prosecutor demanding that someone is brought by force to their country in order to be questioned and that that's not a decision being made by a judge or a court is alarming to us, because we believe in judicial...May 30, 2012 | Story
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Senate Scrambles to Act on Bulk Phone Spying Before It Expires
Ireland Holds Historic Same-Sex Marriage Referendum
Boy Scouts President Calls for End to Ban on LGBT Scout Leaders
Baltimore Grand Jury Indicts 6 Officers in Freddie Gray's Death
"Say Her Name" Protests Nationwide Honor Black Women Killed by Police
Akai Gurley's Partner Sues New York City over Police Shooting Death
Washington: Cop Shoots 2 Unarmed Men Accused of Trying to Steal Beer
Thousands Call for $15-an-Hour Minimum Wage at McDonald's HQ
Guatemalan President Fires Top Officials amid Corruption Scandal
El Salvadoran Archbishop Óscar Romero, Killed by U.S.-Backed Death Squad, to Be Beatified


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Adopting Agriculture As An Occupation
By Geoffrey Riley & Charlotte Duren
Has the tide turned on interest in farming as an occupation?
For decades, the numbers of family farmers declined, and the acreage worked by large agribusiness companies went up.
Now younger people are turning their attention to farms and ranches.
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User/Comment Login
Editor: Here's an original thought for the day.
If you don't have to give up your car because others drive drunk with theirs, why do you have to give up your gun because others commit crimes with theirs?
Editor: The American people deserve a president who will get this great nation of ours back to sanity.
His legacy is:
■ A debt that will cripple our economy, have our future generations stymied.
■ 23 million Americans out of work or not looking. 43 straight months of unemployment over 8 percent.
Where does anyone see Hope or Change with a record of this devastation?
Obama said if he can't get the economy back on track, he would be a one-term president, America, make that happen.
Editor: You must furnish your old driver's license, birth certificate, social security card, marriage license and two documents to prove your present address. How efficient for Florida. Sad and more than that, I would say disgraceful, the Ninth District Court In Arizona ruled illegals don't have to prove citizenship to vote. Does anyone with their brain functioning see anything wrong with this picture?
Editor: Most of us have forgotten we are the "employers" of our government. When was the last time you ever heard an employee giving themselves raises, perks of any kind? Never. Oh, wait a moment. Our government forgets just who their employer is, the American people.
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UW-RF Home > University Communications Home > This Month's News Releases
University Communications
For more information contact firstname.lastname@example.com or firstname.lastname@example.com.
Graduates Step Into the Sunshine After Commencement
MAY 14, 2007--Some 613 undergraduate and 86 graduate students left the commencement ceremony at the Robert P. Knowles Recreation Center Saturday with a perspective of global awareness, service, community and the constant of change as they ventured out into the bright sunshine to start their new lives as alumni of the University of Wisconsin-River Falls.
Commencement speakers Dana Laufenberg, an animal science major from Blanchardville, Wis., and Nicole Peters, a speech communication major from Clear Lake, Wis., were selected to address their peers and the families and friends of the graduates.
"All of us in black robes today have one privilege in common, that of a college education," said Peters, who addressed the afternoon ceremony which included graduates of the College of Arts and Sciences and the College of Education and Professional Studies. "To simply benefit ourselves with this would only slow the momentum of the ever increasing global community we belong to. While it might require sacrifice or striving towards the unknown, we have the ability to serve as active citizens in the global community."
Peters spoke fondly of the many unique relationships she developed while a student. "I have developed close relationships with the residence hall custodians, especially Jean, Mars, Cheryl and Debbie. These women have imparted to me the importance of an attitude of servitude. ... They take pride in their job they do and spread laughter to all they encounter. ... I can only hope that in our future endeavors that we are able to bring such joy and enthusiasm to any role we are placed in and to always live a life with an emphasis of serving others just as these amazing women do daily in the halls."
In her speech, "Journeying Through Life," at the morning ceremony to graduates of the College of Agriculture, Food and Environmental Sciences, the College of Business and Economics as well as the graduate school, Laufenberg told her colleagues, professors and family and friends of the graduates that adaptability to change is a constant.
"We have witnessed the fastest revolution and adaptation to technology the world may ever see," said Laufenberg. "After graduation we will continue to make our own decisions that will impact every aspect of our lives. We will face challenges that will be more and more complicated. ... We will continue to witness changes that will forever impact our lives.
"After today we don't have teachers guiding and updating us," Laufenberg continued. "It is up to us to decide, to choose, to adapt to the changes coming at us, and to continue setting goals that are much higher than [reaching] the cookie jar Mom stashed on the top shelf."
The graduates were welcomed by Chancellor Don Betz as well as Faculty Senate Vice Chair Ogden Rogers, a professor of social work at UWRF, and 2006-07 Student Senate President Joseph Eggers, a junior political science major from Appleton, Wis.
The 2007 Distinguished Teacher, James Madsen, a physics professor, was presented to the graduating seniors and the 2007 Distinguished Alumni Award was presented to Dr. John E. Clemons, a retired physician and 1957 graduate of UWRF.
Degrees were conferred by Provost and Vice Chancellor of Academic Affairs Charlie Hurt, and Dale Gallenberg, dean of the College of Agriculture, Food and Environmental Sciences, Barbara Nemecek, dean of the College of Business and Economics, and Doug Johnson, director of graduate studies at the morning ceremony; and at the afternoon ceremony by Hurt, Terry Brown, dean of the College of Arts and Sciences, and Connie Foster, dean of the College of Education and Professional Studies. Paul Rebholz welcomed each set of graduates into the UWRF Alumni Association.
The collegiate experience remains more than just academics, Peters told the afternoon audience in her speech titled, "Lessons Learned in the Textbook of Life." "The knowledge that we take with us from this institution transcends beyond ... textbook lessons to things we have learning about ourselves from those with whom we have been surrounded by."
A reception was held in the W.H. Hunt Arena for the graduates, their families and friends and faculty and staff.
Last updated:
Thursday, 22-Apr-2010 16:08:19 Central Daylight Time
University of Wisconsin - River Falls


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Latest Rote Island Stories
JAKARTA (Reuters) - An Indonesian ferry carrying around 100 people has sunk in the eastern part of the country, but no deaths have been confirmed so far, rescue officials said on Tuesday.
JAKARTA (Reuters) - The death toll from a ferry sinking in Indonesia this week has reached 10, while 125 people have been rescued, a search and rescue official said on Friday.
JAKARTA (Reuters) - An Indonesian ferry carrying 105 people has gone missing in heavy seas in the country's east and naval vessels had been called in to search for it, a port official said on Wednesday. El Shinta radio station said the vessel had sunk.
Latest Rote Island Reference Libraries
The Timor Monitor or Spotted Tree Monitor, Varanus timorensis, is a species of small lizard native to Indonesia, New Guinea and Australia. More specifically they are found on the islands of Timor, Savu and Rote in Indonesia. They are found in Samoa in southern New Guinea, and in North territory, Queensland, West Australia. The Timor Monitor is generally a dark green or almost black in color, with bright gold-yellow spotting all along its back and a lighter straw-yellow coloring on its...
- Stoppage; cessation (of labor).
- A standing still or idling (of mills, factories, etc.).


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What does it mean? For the answer, check this book
The Servant: A Simple Story About the True Essence of Leadership
I discussed it a little in this post The Spiritual r3VOLution Inspired by Ron Paul
How can you love an ax murderer, after all? It was in The Servant that I found the distinction of what this means. The Greeks had five words for love: eros, erotic love; storgé, love for family members; philos, brotherly / reciprocal love and finally, agapé – an unconditional love rooted in behavior towards others without regard to their due. In other words, do unto others as they would have them do unto you, regardless of how you feel about them personally.
What a blunt instrument the English language is, having only one word to cover so many distinct concepts!
When Jesus speaks of 'love' in the New Testament, he speaks of agapé, the love of behavior and choice, not of feeling. In this sense, to love your neighbor or your enemy (agapé) means to show them the same patience, kindness, humility, respect, selflessness, forgiveness, and honesty – regardless of whether you agree with them or not. Just as Dr. Paul does. He treats everyone – friends and opponents alike – with the same patience, kindness, humility, respect, forgiveness and honesty that constitute agapé love.
That is just a taste of what you'll find in the book, which I'm rereading now.
Thank you for reading.
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CALDWELL – The Blackfoot Broncos rode the right foot of senior Gerardo Bartolo all the way into the 4A State Soccer Championship game. The talented striker knocked in not one, not two, but all three Blackfoot goals in a hard fought 3-2 victory over Skyview on Friday afternoon at Caldwell High School.
"Right from the beginning of the game you could tell that we were all focused and ready to go," Bartolo said. "We want to win a championship so badly and we came out attacking, attacking and just taking the game to them. To get three goals, my senior year, well that feels amazing."
CALDWELL – The Snake River Panthers took a big step towards a 3A state championship on Friday afternoon as they turned in a convincing performance against perennial soccer power Payette with a 6-1 victory.
"The guys just came out and killed it today," Snake River head coach Trevor Peterson said. "They came out and played an awesome game and I couldn't be happier."
Snake River drew first blood in the game in the eighteenth minute of play when a Payette player was called for an intentional hand ball on a ball that was heading for an open goal on a corner kick.


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Chiarelli: Bruins 'cooperating fully' with league regarding Savard
|08.10.10 at 3:53 pm ET
The Bruins have released a statement from general manager Peter Chiarelli regarding the league's investigation of the seven-year, $28.05 million contract extension center Marc Savard signed with the team in December. Here's what Chiarelli had to say.
'We are cooperating fully with the League in its investigation of the Marc Savard contract extension. The League informed us upon their registration of the contract on December 1, 2009 that they would be investigating the circumstances surrounding this contract. From that point on, they commenced their investigation and it has been ongoing since then. On August 4th, I met with two League appointed lawyers as part of the investigation. We will continue to cooperate with the League in any future investigative proceedings if necessary and we will have no further comment on the matter at this time.'
Savard will make nearly half of his contract's money in the next two seasons, with the other half being spread out over the following five seasons. As a result, despite making $7 million per year in 2010-11 and 2011-12, he will have a cap hit of just $4.007 as a result of cap numbers in the NHL being determined by total money divided by years. If the deal is ruled as not being in compliance with the CBA, it will be voided and Savard will become an unrestricted free agent.
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AFI FEST 2003, opening today, still lurks in the shadows of internationally renowned film festivals such as Toronto, Berlin, Venice and Sundance, but it is gaining industry respect and audience notice as a showcase for award-worthy titles and cinematic treasures from Europe, Asia and Latin America.
``Beginning last year and this year, I think it's moving in the right direction, because it's trying to be a true international film festival, a festival that brings in films from around the world,'' said film critic Henry Sheehan, who has attended festivals around the globe for about 20 years.
Sheehan, who reviews films for KCET and KPCC-FM as well as online at henrysheehan.com, said the American Film Institute's yearly showcase has improved greatly since festival director Christian Gaines set its focus on international works.
``They had that bad period where they tried to be an American independent film festival and that was a terrible idea for the AFI,'' Sheehan said. ``It's not a terrible idea for Los Angeles because the Los Angeles Independent Film Festival does it well, but the AFI Fest wants to be the major festival for the second largest city in the country and the only way to do that is to be an international festival.''
The 17th annual AFI Fest, running through Nov. 16, has 14 films in the International Feature Competition, among them the U.S. premiere of ``Nicotina,'' a comedy production of Mexico, Argentina and Spain and starring Diego Luna of ``Y Tu Mama Tambien.''
There are 12 projects in the International Documentary Competition, eight in Latin Cinema, eight in Asian New Classics, 12 in the European Showcase and six in Beyond Conflict: Focus on the Middle East. The 10 titles in its Made in Germany section include a new Robert Fischer documentary about Hollywood's influence on the work of German filmmaker Rainer Werner Fassbinder, titled ``Fassbinder in Hollywood.''
``Providing Angelenos a festival of record for international films - something that people can step into year after year and think, 'I know this festival' - that's paramount for us,'' Gaines said.
Festival watchers have praised AFI's decision to set up the entire film series at ArcLight Cinemas in Hollywood, which combines the glam appeal of the 40-year-old Cinerama Dome with a new multiplex, gracious reception areas and, most important in Los Angeles, easy parking.
It paid off in 2002, with attendance of some 40,000, up about 33 percent from the previous year when moviegoers shuttled among four Hollywood venues. Regular attendees even noted the greater diversity of the audience, which could be attributed to wider marketing and the appeal of its international titles.
Sheehan sees funding as AFI Fest's greatest hurdle toward gaining importance on the world cinema scene. He suggested more money from Los Angeles-based businesses such as real estate developers, law firms and fashion companies would allow Gaines more manpower and travel funds dedicated to seeking out the best new international films and setting AFI for their world or U.S. premieres.
More money also would feed the publicity animal, which lures not only moviegoers but celebrity actors and filmmakers and would in turn attract more high-end sponsors.
``You have to replace the money you spend on a year-to-year basis - that's the bare minimum,'' Gaines said. ``Corporate sponsorship is a critical component and a main funding source for the festival, which offers brand awareness and visibility in exchange for value. Ticket and pass sales is a rapidly growing critical arm of our funding.
``It's always a struggle, it's always a challenge,'' he said. ``I spend a lot of time on it, as do a lot of people here. It's just a part of the work.''
With the exception of some deals brokered in the Kodak Connect center, Gaines does not wish AFI Fest to become a market for distribution deals as Cannes and Sundance are.
``The last thing we want is to spring a market on the film community with all the pressure and anticipation that engenders,'' he said.
But the AFI has taken a step toward merging the creative and commercial sides of the industry with an alliance with the annual American Film Market, which next year will shift its annual gathering from February to November, abutting the AFI Fest dates on the calendar. ``Even if there is a very limited cross section of participants involved in both, I think it will be a very active, very intensive cross section,'' Gaines said.
Sheehan praised Gaines and his staff (six year-round, growing to about 30 paid positions at festival time) for ``doing a really good job in the face of no support.''
``If a city wants to have an international status as an arts capital it absolutely needs to have a respectable film festival,'' Sheehan said. ``Unless these businesses stand up and put their money there, Los Angeles isn't going to have one.''
Gaines said when he joined AFI Fest in summer 2000, his colleagues in the film fest trade suggested it would be a tough assignment.
``Everybody said, 'Good luck running a festival there. The L.A. audience is a cynical audience and no one wants to go to anything unless it's free or there are a bunch of celebrities.'
``But it's not just a company town,'' Gaines said. ``It's turning into a great festival town. The appetite for specialty cinema is changing for the better.
``We went from wondering how we were going to get people into theaters to wondering how we're going to deal with crowds, and that's been a major turnaround.''
AFI Fest has launched some sleeper hits of recent awards seasons, including ``Monster's Ball,'' ``You Can Count on Me'' and ``The Cider House Rules.''
The buzz this year is mostly whirling around three films:
Robert Altman's ``The Company'' examines the grittier side of artistic creation in a story of a ballerina (Neve Campbell) as she goes through her paces under a tough taskmaster (Malcolm McDowell) (7 p.m. Nov. 15).
``House of Sand and Fog,'' with Oscar winners Ben Kingsley and Jennifer Connelly, involves two people embroiled in a fight over ownership of a house (7:30 p.m. Sunday).
``The Statement'' stars Tilda Swinton and Oscar winner Michael Caine. Directed by Norman Jewison, (``In the Heat of the Night,'' ``The Hurricane'') and based on a Brian Moore novel, the film involves a war criminal who had lived a quiet, anonymous life for decades before being put in the spotlight by a new investigation (8 p.m. Wednesday).
But the next awards-season darling or a sleeper hit may as easily be lurking elsewhere in the festival schedule, awaiting a few rave notices and good word-of-mouth rippling through the AFI party scene. There are also a number of offbeat films that are already drawing interest from cinephiles. Here are some you might want to check out:
``Calendar Girls,'' which kicks off the festival tonight, stars Helen Mirren and Julie Walters and is based on a true story of a group of Yorkshire women (7:30 tonight).
``The Fog of War,'' the new documentary from Errol Morris (``The Thin Blue Line''), examines the rise of Robert McNamara, JFK and Lyndon Johnson's secretary of defense and one of the architects of the Vietnam War.
The industry's fresh obsession with Florida murderer Aileen Wournos is the subject of ``Monster,'' the sold-out closing-night gala with Charlize Theron in the title role (8 p.m. Nov. 16). Wournos is also the subject of director Nick Broomfield's ``Aileen: Life and Death of a Serial Killer,'' his second documentary on her (9:45 p.m. Friday).
``In America,'' director Jim Sheridan's semi-autobiographical story, follows an Irish family that immigrates to New York City following the death of an infant son. Samantha Morton and Paddy Considine play the parents of scene-stealing sisters Sarah (10) and Emma (6) Bolger (7:20 p.m. Friday).
``The Triplets of Belleville'' is a French animated production with quirky characters and big musical numbers (7:45 p.m. Wednesday).
``Frankie and Johnny Are Married'' takes a surprisingly charming look at self-deprecating TV producer-director Michael Pressman's near crash- and-burn experience with Equity-waiver theater, which could have cost him his marriage to his leading lady (10 p.m. Saturday and 3:20 p.m. Monday).
``The Barbarian Invasions/Les Invasions Barbares,'' Canada's submission for Best Foreign Language Film, is from director Denys Arcand (``Love and Human Remains''). It chronicles a man's return home to his ailing father's bedside, which is surrounded by relatives, friends and former mistresses (6:30 p.m. Saturday).
In ``Japanese Story,'' Toni Colette stars as an ambitious geologist who suddenly finds herself having to baby-sit a taciturn Japanese businessman. Hoping to strike up a business deal, she agrees to take him on a field trip to Australia's remote Pilbara desert, where they leave more and more of what they know about each other and themselves behind (9:15 p.m. Nov. 15).
Actor/director Jon Favreau (``Elf'') plays an out-of-work actor trying to make sense of such disparate elements as a blue suitcase, 75 missing people, a neighbor who loses his head and a cryptic cowboy in ``The Big Empty'' (7 p.m. Sunday and noon Monday).
All screenings are at the ArcLight Hollywood (6360 Sunset Blvd.), with the exception of the Omar Sharif retrospective, which is playing at the Skirball Cultural Center (2701 N. Sepulveda Blvd.). General admission tickets are $11. Passes range from $95 to $500. Nonpass tickets to the opening and closing galas are sold out, but last-minute rush tickets may be available. For information: (866) 234-3378 or www.afi.com/afifest/2003.
Valerie Kuklenski, (818) 713-3750
5 photos, box
The Art of a Film festival
(1 -- cover -- color) `THE STATEMENT'
WHAT'S PLAYING (see text)
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|Nov 6, 2003
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text: The mighty Ohio River has been running high and fast these days thanks to all the precipitation in all its forms this winter. The river crested at Point Pleasant Riverfront Park on Wednesday, Feb. 5 at 30.34 feet, according to the National Weather Service - flood stage is 40 feet. Still, this was enough to bring the water well into the amphitheater, along with mud and debris. As of 5 p.m. on Thursday, the water had receded to 29.65 feet. No doubt the Ohio River will be back again as this winter marches on and into a chance of more rain and snow expected this weekend.


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Woah. We knew that the Weinstein Company were about to unveil some footage from Paul Thomas Anderson's "The Master" in Cannes tonight (and we'll have our reactions from that for you later), but we just got a nice surprise, courtesy of the Twitter account of Megan Ellison, with the first footage from...Read More »
Here's something to whet your appetite for the fall film festival season. While journos, pundits and fans alike all lamented the absence of Paul Thomas Anderson's "The Master" in the Cannes line-up this year, it appears The Weinstein Company -- already dominating the fest...Read More »
You've got to assume the secretive folks at Annapurna Pictures are fairly amused this afternoon after opening the latest edition of the New York Times. Inside, is a curiously timed, rather headscratching article about Paul Thomas Anderson's upcoming drama, expected to be called, "The Master."Read More »
There's good news and bad news tonight. The potentially bad news is that the glut of commentors who insist that Paul Thomas Anderson's upcoming drama, "The Master" isn't actually called that are wrong according to Box Office Mojo. The good news, as promised by Annapurna Picture...Read More »
Way back in April, before lensing started on Paul Thomas Anderson's "The Master," word leaked out that the director was shooting test footage with a 65mm camera, the same used by Stanley Kubrick on "2001: A Spacy Odyssey" -- and then, not much was heard after that. Well, it looks like those tests we...Read More »
In a time where there seems to be a constant focus on set photos and early looks, it's reassuring to see distinguished auteur Paul Thomas Anderson's next untitled offering (aka "The Master") make its way to theaters so quietly.Read More »
As much a film scholar as a filmmaker, there are few directors better-equipped to discuss the convergence of art and technology than Martin Scorsese. But during a recent Q&A about his latest film, the 3D opus "Hugo," Scorsese offered a few observations about the past, present and fut...Read More »
'Drive' Star Also Passed On 'Big,' 'Dead Poets Society' And 'Pretty Woman'Of the many things that have made a comeback in 2011 -- Wilson Phillips, high-waisted pants, the street protest -- perhaps the happiest is the resurgence of Albert Brooks. Mostly absent from screens in the past decade, bar a vocal turn in Pixar's masterpiece "Finding Nemo," and his directorial flop "Looking For Comedy In The Muslim World," Brooks returned with an acclaimed book, "2030: The Real Story of What Happened to America," took to Twitter and instantly became the funniest thing on it, and played, against type, mobster Bernie Rose in Nicolas Winding Refn's "Drive,...Read More »
It's an early sign of talent to watch out for when little-known actors manage to score roles in major, highly anticipated films. Going by that theory then, Ambyr Childers -- best known for her stint as Colby Chandler on "All My Children" -- is definitely someone to keep an eye on after nabbing a par...Read More »
Precariously mixing elements of broad comedy, bittersweet heartache, charming romance and soulful, surprisingly mature dramatic moments, "Crazy, Stupid, Love" is an endearing and winning summer comedic drama. It succeeds in spite of some messy plotting, dicey adulteration of tones, and too convenien...Read More »


text:
The Boy Scout slogan of do a good turn daily is something ingrained into the heart of every Scout from the very start of his journey. The slogan sets kindness and willingness to go above and beyond to help those in need as the foundation for the rest of Scouting. Despite this important mission of lending a hand to those without a voice, the Scouting experience has been denied to those who are openly homosexual.
The time spent in meetings, around campfires, overcoming hardships and helping each other along the Scouting path is what creates the valued qualities scouting instills. It is not merely the memorizing of mottos, oaths and slogans that develops the boys who enter Scouting into the young men who leave it, but through the common experiences they share with each other along the way. The current policy of exclusion not only hurts the boys excluded, but also limits those in Scouting. By limiting who can join, this policy stifles chances for a Scout to learn about someone different than him, to engage someone in a discussion that can change ideas from an unknowing fear into a friendship.
The Boy Scouts of America, in opening a discussion to change the current policy excluding homosexual Scouts and leaders, is taking a step towards expounding on its fundamental principles and molding responsible and active leaders in the community. This is a discussion I hope ends with the inclusion of more young men who can use the values instilled by the Boy Scout experience to grow to become dependable and dedicated leaders in their communities.
What ever happened to Luzerne county's number one veteran advocate?
A few months ago I was sitting at my local VFW complaining about all the crap I was going through trying to get my VA compensation claim completed. As a veteran of Afghanistan and Iraq, I couldn't believe what I was going through; the paperwork, red tape and the misinformation. A buddy of mine suggested I call Rick Wren, former director of Luzerne County Veterans Affairs. He said if anyone could help me, Wren could. He also said Wren helped him with his claim.
I called Mr. Wren on a Sunday and first apologized for bothering him and told him what I was going through. He graciously told me he would help me any way could.
Over the next two hours he took me through the whole VA process. He explained how the system works and what I needed to do. Here is a man who took the time to put me on the right track and received nothing but my thanks.
The wealth of knowledge he demonstrated astounded me and he was able to explain it all into words I could understand.
Yesterday I received a letter from the VA and it appears I will finally be getting my award. I called him and thanked him again and he said to me no thanks were needed. He said it was the least he could do for the sacrifices I endured for him, his family and the country. I had to write this letter to let the veteran community know that Rick Wren is alive and well and still taking care of veterans.
Mr. Wren, thank you and God bless you and your family. I could not have gotten my award without you.
John Dougherty Hazleton Stewart Long Mountain Top


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How to Scan Transparency
There is a chance that while collecting your family negatives or slides, you receive a bunch of negative or slides. You might be more interested to include those slides into your family history or you want to publish those images on the website or you want to make them as digital copies and want to share with your family members.
Do you ever try to scan the slides or negatives on flatbed scanner? If yes, then you will know that they do not work. The output of this scanning is a big black square. In this article, I'm going to explain how to scan transparency and what are the issues that you will face.
The key benefits of the scan transparency:
Here, you will first get to know about the benefits of scan transparency and then about scanning a photo. When you are using scan transparency, you can use total area of negative or slide whereas in photo scanning, it will provide the same version of the photo. In the process of converting slide film into positive, you have to insert the colors into that and remove the orange shade from the negative.
The flatbed scanner is designed for photo scanning and other images scanning. The main problem with this scanner is the light that passes through the image rather than reflected off image. If you want a successful scan transparency then you have to use scan transparency adapter.
For perfection, a scan transparency adapter is not a best solution for this problem because your flatbed scanner contains a piece of glass between the sensor and the negative.
Other than flatbed scanner, DPI or Digits per Inch resolution is around 600 and the interpolated DPI resolution is 1200. Let your flatbed scanner has 600 DPI resolution then it will produce 4x4 inches picture with good quality of resolution that means half of the letter – sized paper. The calculation is like, if you are printing 1 inch at 600 DPI, which means 1x600 pixels, then the total is 600 x 600 pixels. The output resolution is 150 DPI that is 600 / 150 = 4 inches. If you are printing at high output resolution then the final image will be reduced very quickly. For example, if you are printing at 600 DPI and the output resolution is 300 DPI then it will reduce to 2 inches that means 600 / 300 = 2 inches.
Based on this calculation, your flatbed scanner with scan transparency adapter will provide focus on the negative or film. In some scanners, the focus is very less and so the result is less sharp than the original one. If you want sharp scan then remove the slide from the mount.
If you want better scanning with high resolutions and high quality then get film scanner, which is also called as slide scanner used for scanning slides and negatives. In this, you need not have to use extra scan transparency. The DPI resolution of this scanner is 2400 or high. Once you created positive from negative, use photo-editing tools to modify the image.


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Welcome to the first Marvel Press Release of 2013!
As with Marvel's usual cryptic messages we can speculate all we want about what this title will be and who will be involved but I'm gonna put my money on this NOT being an all female team, but simply the transition of either the appropriately self titled "X-Men" or "X-Factor" to the new Marvel Now! logo. Both titles aren't exactly blowing anyone away and they are in serious need of a kick-start. What better time than April? Here is the official press release.
The Future of Marvel NOW! Is XX.
Brian Wood.
Olivier Coipel.
April 2013


text:
Secondary menu
Although a majority of these theories are far-fetched, some make the movie they're about feel more relatable and understandable.... READ ON
A few things you might not know about Marty, Doc, and Doc's pet chimpanzee.... READ ON
Looking for George Lucas and Steven Spielberg's easter eggs.... READ ON
Plenty of the Great White Way's most notable productions have been reincarnated for the silver screen—think Rent, The Producers, and Chicago—but Off-Broadway and Broadway's best have also spawned plenty of big screen versions that most people don't realize started as a live theatrical experience.... READ ON
They're presented in reverse order, so you can quiz yourself as you go along.... READ ON
Hollywood is changing: Paramount Pictures no longer produces films on celluloid and Old Guard directors like Martin Scorsese are shooting their movies with digital cameras. Here's a list of important films that helped propel Tinseltown's controversial shift from film into digital, from niche-market fad to high-profile industry standard.... READ ON
It's often the first thing you know about a film, so when it comes to crafting the next blockbuster, movie makers are eager to get the name just right. That means if the first—or second—suggestion isn't a winner, it's back to the drawing board to find something pithier, pluckier, or otherwise more appealing. Here's a look at some of the movie titles that didn't make the cut.... READ ON
Hollywood pushes out a seemingly endless supply of movies based on novels, comic books, or old TV shows. Every so often, however, studios try their hand at adapting poems for the big screen. Here are 11 examples.... READ ON
It's not easy to become a star, but here are some proven short cuts from the past 100 years of movie stardom.... READ ON
E.B. White of Charlotte's Web fame is the "White" of Strunk and White.


text:
Tallon Green
from £51 /night help Price for guests, Nights approx:
from £51 /night help Price for guests, Nights approx:
Estimated nightly price based on a weekly stay. Contact the advertiser to confirm the total cost.
Tallon Green hidden in a quiet corner of picturesque Litton village in the heart of the Peaks.
Cottage / 2 bedrooms / sleeps 4
Availability Your dates are available
Cottage / 2 bedrooms / sleeps 4
|Key Info
Tallan Green Cottage is an utterly wonderful cottage with spacious accommodation that has been superbly decorated and furnished to create the ultimate holiday home. The luxurious sofas and glowing stove in the fantastic lounge, ultra modern kitchen, superb bedrooms with quality linen and the classy bathroom, just about say it all. Then remember that the village pub is just round the corner.
Litton with its 'real' local shop and famous Red Lion pub is a very picturesque village in the midst of fantastic Peak District scenery. There are walks galore in every direction but best of all are dramatic Millers Dale or Cressbrook Dale running into beautiful Monsal Dale where, if you are lucky, you can share your picnic lunch with the ducks as you marvel at the rainbow in the sunlight over the lovely waterfall.
Surrounded by all this magnificent scenery you will be lost for words if you are walkers but don't forget that Chatsworth, Bakewell and all the other famous sights are close by. Our local favourites include listening to the incredible echo in the Dome in Buxton, reliving the Dambusters on a visit to the Derwent Valley Dams and exploring the amazing Blue John caverns in Castleton.
Cottage facilities.
A superb and spacious beamed lounge with magnificent stone fireplace, glowing coal effect gas stove, luxurious sofas, flat screen T.V, DVD and SKY. A wonderful stone flagged dining room with under floor heating and mellow pine table and chairs. An ultra modern kitchen with quality units, hob, oven, microwave, fridge-freezer, dishwasher and automatic washer. A classy double bedroom. An attractive twin bedroom. A nursery room childs single bed. A spacious 'state of the art' bathroom with bath, separate large corner shower and W.C.
Linen and towels are provided. Central heating, fuel and power are included. A cot and high chair can be requested. There is a little front patio with bench. Parking is on the quiet lanes around the village within 50 yds.of the cottage. Pets and smoking are not permitted. Access for people with disabilities may be difficult.
Nearest pub - Litton 100 yds. Nearest village shop - Tideswell 1.mile. Nearest supermarket - Buxton 5 miles.
Week bookings are normally from Saturday to Saturday. Short breaks are available between November and March (excluding Xmas and New Year) and during the rest of the year if the booking is within 28 days of the holiday. The minimum short break price is for three nights.
|Sleeps up to 4, 2 bedrooms
|Check in time:
|Check out time:
|Will consider
|Long term lets (over 1 month)
|Car advised
|Nearest Amenities
|2.5 km
|Nearest travel links
|Nearest airport: Manchester 40 km, Nearest railway: Buxton 15 km
|Family friendly
|Great for children of all ages
|No pets allowed, No smoking at this property
Features and Facilities
|DVD player
|Central heating, TV, CD player, Satellite TV
|Kettle, Toaster
|Clothes dryer, Dishwasher, Cooker, Microwave, Fridge, Freezer, Washing machine
|2 bedrooms, 1 bathrooms of which 1 Family bathrooms
|Double beds (1), Single beds (2), Cots (1), Dining seats for 6, Lounge seats for 6
|Linen provided, Towels provided, High chair
|Balcony or terrace
The Peak District National Park region
Derbyshire is in the centre of the country and a brilliant place for a countryside holiday with its beautiful landscape that inspired Britain's first National Park. The scenery is fantastic with picturesque villages, sparkling rivers in lovely valleys and high moorland areas where you will feel 'on top of the world'. The views over wave after wave of Peak District hills are just incredible !
Walkers will find lots of lovely walks. The trails out across the 'Peaks' are great for cycling. There is super heritage with numerous stately homes to explore. And numerous holiday attractions can be found in and around the 'Peak' as well as great days out at nearby Alton Towers or Robin Hood's Sherwood Forest .
On less strenuous days you might fancy at bit of shopping. The big shopping centres of Derby, Sheffield, Nottingham and Manchester are easy to get to. Local towns like Ashbourne, Bakewell and Buxton have lots of individual country town shops. Take home a bargain from one of the many factory outlets.
When you are out and about remember that there are many classy restaurants in the county. Country pubs serving super food and real ale abound. You might just fancy a picnic from one of the little deli's - every town has got one. There is also a lot to be said for a super evening takeaway in the cottage. Food and drink to suit every taste and budget !
Finally lets mention a few local secrets. Derbyshire's famous well dressings must be seen. Ashbourne's Royal Shrovetide Football is amazing. Majestic Chatsworth House is the star of all the stately homes. Carsington Water's wild water sports are a must. Bakewell's famous Monday market is well worth a visit. Matlock Bath's 'little Switzerland' gorge and Heights of Abraham cable cars can be heart stopping! Buxton's Opera House and the echo in the Devonshire Dome are incredible.
But the best thing of all about Derbyshire is that it is a fairly small place and everything is very accessible. There is something within easy reach to fill every day of your holiday ! Please give us a call if you are still stuck for ideas. Hope you come back next year to finish the experience !!
There is an incredible variety of things to do and see in Derbyshire and the Peak District to fill every day of your holiday.
People who love heritage will be enthralled by the Derwent Valley World Heritage Corridor and the numerous stately homes such as Chatsworth House, Haddon Hall, Hardwick Hall and the time capsule of Calke Abbey. Walks by the old Cromford and Whaley Bridge Canals are great and a visit to Creswell Crags is a must.
There are lovely walks all over the county but try climbing Mam Torr as the para-gliders float by, explore the Derwent 'Dambuster' Dams, start the Pennine Way in wild Edale, wander down peaceful Dove Dale, Manifold Dale, Monsal Dale or Lathkill Dale. And don't forget the breathtaking views from Calver Edge!
Holiday attractions abound with Carsington Water, the Peak, Midland and Ecclesbourne steam railways, the Heights of Abraham cable cars, Castleton's Blue John Caverns, the National Tramway Museum, the Chestnut Owl and Otter Sanctuary and even nearby Alton Towers and Robin Hood's Sherwood Forest.
For active days out it's got to be the water sports at Carsington, cycling along the High Peak and other trails, climbing the cliffs or canoeing the river in Matlock Bath, fishing the numerous rivers, playing a round of golf at one of the lovely courses, gliding at Gt. Hucklow or even go-carting in Buxton or Chesterfield.
Special events happen throughout the year so there is always something exciting happening. Matlock's Boxing Day raft race is an amazing spectacle. Royal Shrovetide Football in Ashbourne is an incredible event. All the famous village well dressings are beautiful. Buxton's opera season is superb. Chatsworth's Country Fair draws crowds from far and wide. Bonsall's hen racing is unique and Fenny Bentley's toe wrestling is bizarre. The list goes on and on!
If you fancy a night out there are night clubs in Chesterfield and Derby, theatres in Buxton, Chesterfield and Derby, multi-plex cinemas in Chesterfield and Derby and lovely little local cinemas in Belper and WIrksworth. But most people will just like to try one of the numerous restaurants and pubs catering for every taste and budget.
Our favourite restaurants include the Devonshire Arms in Beeley, Stones' little riverside bistro in Matlock, Simply Thai in Buxton, Fellicinis' Italian in Bakewell, Le Mistral in Wirksworth, Rowley's bistro in Baslow, the Plough in Brackenfield and, for a very special occasion, fine dining at Hassop Hall. Don't forget to have a Bakewell Pudding before you leave!!
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Sylvia Stimson (Property Manager Derbyshire Country Cottages)
- 1 Years listed
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Calendar last updated:18 May 2015
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