text:
Rapid identification of bacteria by mass spectrometry
A method for the identification of bacteria using genus, species and strain specific biomarkers. The biomarkers are generated by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis of either cellular protein extracts or whole cells. Biomarkers for a number of pathogenic bacteria are disclosed.
Krishnamurthy, Thaiyalnayaki (Ellicott City, MD)
Ross, Philip L. (Jessup, MD)
The United States of America as represented by the Secretary of the Army (Washington, DC)
435/173.1
250/281, 250/282, 250/287, 313/564, 435/173.4, 436/173, 436/518
H01J49/04; (IPC1-7): C12N13/00
436/518, 436/173, 250/281, 250/282, 250/287, 435/173.1, 435/173.4, 313/564
Krishnamurthy et al: "Detection of Pathogenic and Non-Pathogenic Bacteria by Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry". Rapid Communications in Mass Spectrometry vol. 10, 883-88 (1996).
Krsihnamurthy et al: "Rapid Indentification of Bacteria by Direct Matric-assisted Laser Desorption/Ionization Mass Spectrometric Analysis of Whole Cells", Rapid Communications in Mass Spectrometry vol. 10, 1992-1996 (Dec. 1996).
Krsihnamurthy et al; "Bacterial Detection by Mass Spectrometry". Indian Society for Mass Spectrometry pp. 105-122 (Nov. 1996).
Zaluzec et al; "Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Applications in Peptide and Protein Characterization". Protein Expression and Purification vol. 6, pp. 109-123 (1995).
Holland et al; Rapid Indentification of Intact Whole Bacteria Based on Spectral patterns using Matrix-assisted Laser Desorption/Ionization with Time-of-Flight Mass Spectrometry. Rapid Communications in Mass Spectrometry vol. 10, 1227-1232 (Jul. 1996).
Cain et al; Differentiation of Bacteria Using Protein Profiles from Matrix-assisted Laser Desorption/Ionization with Time-of-Flight Mass Spectrometry. Rapid Communications in Mass Spectrometry vol. 8, 1026-1030 (1994).
Harvey; "Matrix-assisted Laser Desorption/Ionization Mass Spectrometry of Oligosaccharides and glycoconjugates". Journal of Chromatography a 720 (1996) pp. 429-446.
Chevrier et al. Matrix-assisted Laser Desorption/Ionization with Time-of-Flight Mass Spectrometry Based on a 600 ps. 1.2mJ Nitrogen Laser. Rapid Communications in Mass Spectrometry vol. 5, 611-617 (1991).
DRDE, Gwalior; "Proceedings of Seventh National Symposium on Mass Spectrometry". Indian Society for Mass Spectrometry (ISMAS) Mumbai, India (1996).
Claydon et al.; "The Rapid identification of intact microorganisms using mass spectrometry". Nature Biotechnology vol. 14, pp. 1584-1586, Nov. 1996.
Liang et al.; Determination of Bacterial Protein Profiles by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry with High-performance Liquid Chromatography. Rapid Communication in Mass Spectrometry, vol. 10, pp. 1219-1226 (1996).
Graser, Jennifer
Biffoni, Ulysses John
Ranucci, Vincent J.
Parent Case Data:
This application is a continuation of application Ser. No. 08/847,419, filed on Apr. 24, 1997, now abandoned, which in turn is a nonprovisional continuation of provisional application Ser. No. 60/016,438, filed on Apr. 29, 1996.
1. A method for generating biomarkers specific for a known genus, species, or strain of bacteria comprising:
(a) mixing a bacteria sample comprising a suspension of whole cells of known genus, species or strain with a matrix solution to generate a sample mixture;
(b) placing an aliquot of said sample mixture on the probe tip of a time-of-flight mass spectrometer and allowing it to dry;
(c) irradiating the dried aliquot with a nitrogen laser to form ionized biopolymers;
(d) accelerating the ionized biopolymers by an electric field toward a detector through the flight tube of the time-of-flight mass spectrometer to obtain a mass spectra;
(e) averaging the mass spectra resulting from 100 to 200 laser pulses;
(f) repeating steps (a)-(e) with at least one other, nonidentical bacteria sample comprising a suspension of whole cells of the same genus, species or strain;
(g) comparing the averaged mass spectra obtained for each bacteria sample;
(h) identifying at least one ion that is common to each bacteria sample; and
(i) assigning an m/z measurement of the ion as a genus, species, or strain specific biomarker.
2. The method of claim 1, wherein the matrix solution comprises about 10 mg/ml of compounds selected from the group consisting of 4-hydroxy-α-cyano-cinnamic acid and 3,5-dimethoxy-4-hydroxy cinnamic acid in an aqueous solvent solution.
3. The method of claim 2, wherein the aqueous solvent solution for making matrix solutions comprises 0.1% aqueous trifluoroacetic acid (TFA) and acetonitrile in a ratio of 70/30 (v/v).
4. The method of claim 1, wherein the sample mixture comprises 1 μl of the bacteria sample and 9 μl of the matrix solution.
5. The method of claim 1, wherein the time-of-flight mass spectrometer is operated at 26 kV acceleration voltage in linear mode.
6. The method of claim 1, wherein the time-of-flight mass spectrometer is equipped with a low mass ion gate which rejects ions below 1500 Daltons.
7. A method for determining the genus, species and/or strain of an unknown bacteria sample which comprises:
(a) generating an averaged mass spectra according to steps (a)-(e) of claim 1; and
(b) comparing the averaged mass spectra of the unknown bacteria sample to a plurality of genus, species or strain specific biomarkers, said biomarkers being generated according to claim 1.
9. The method of claim 8, wherein the aqueous solvent solution comprises 0.1% aqueous trifluoroacetic acid (TFA) and acetonitrile in a ratio of 70/30 (v/v).
10. The method of claim 7, wherein the sample mixture comprises 1 μl of the bacteria sample and 9 μl of the matrix solution.
11. The method of claim 7, wherein the time-of-flight mass spectrometer is operated at 26 kV acceleration voltage in linear mode.
12. The method of claim 7, wherein the time-of-flight mass spectrometer is equipped with a low mass ion gate which rejects ions below 1500 Daltons.
13. A biomarker library for identifying the genus, species and/or strain of an unknown bacteria sample, the library comprising genus, species or strain specific biomarkers for known bacteria generated by the method of claim 1 using bacteria selected from the group consisting of: Bacillus anthracis, Bacillus thuringiensis, Bacillus cereus, Bacillus subtilis, Yersinia pestis, Francisella tularnsis, and Bucella melitensis.
The present invention relates generally to the rapid identification of bacteria in environmental and biological samples. More specifically, the present invention relates to a method for the chemotaxonomic classification of bacteria with genus, species and strain specific biomarkers generated by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis of either cellular protein extracts or whole cells.
Rapid and accurate microbial identification is critical in diagnosing diseases, predicting on-coming public health hazards, monitoring potential contamination in stored foods and grains, regulating bioprocessing operations and recognizing warfare threats. It is important not only to rapidly distinguish between related organisms but also to unambiguously identify species and strains in complex matrices for risk assessment in field situations.
The classification of micro-organisms has traditionally been based on biochemical and morphological culturing tests. Recently, several instrumental analytical techniques have been developed which enhance the speed and accuracy of identification of bacteria cells. In these techniques, the biochemical components of bacteria cells are examined to determine chemotaxonomic markers which are specific for each bacteria species. The chemotaxonomic markers, or biomarkers, may be any one or a combination of the classes of molecules present in the cells such as lipids, phospholipids, lipopolysaccharides, oligosaccharides, proteins and DNA.
For example, a commercial microbial identification system uses gas chromatographic analysis of fatty acid methyl esters (Microbial Identification, Inc., Newark, Del.). Chemotaxonomic identification of bacteria based on fatty acid or whole-cell pyrolysis mass spectra and fast-atom bombardment mass spectrometric analysis of phospholipids has also been reported. These techniques analyze primarily the lower molecular weight lipids of the cell.
Bacteria may also be differentiated on the basis of cellular protein content. Since the proteins found in bacteria provide indirect genetic information on the organism and are related to bacterial virulence, protein content is specific to individual strains. The most established technique for examining cellular protein content is sodium deodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) which produces characteristic migration patterns for different bacteria species. Identification of a bacteria producing a particular migration pattern is accomplished by computerized comparison with reference gel patterns. However, because SDS-PAGE analysis is slow, labor intensive and requires fairly large amounts of sample material, it is not particularly useful for rapid identification of bacteria, particularly in field situations.
A limited number of recent reports have investigated the applicability of various mass spectrometric techniques for generating bacteria specific protein profiles. These techniques generally employ electrospray ionization (ESI) or matrix-assisted laser desorption ionization (MALDI) of bacteria protein extracts followed by mass spectrometric (MS) or tandem mass spectrometric (MS/MS) analysis. For example, the MALDI technique, combined with time-of-flight mass spectrometry (TOF-MS), has been used to differentiate bacteria using a crude protein extract requiring minimal sample preparation. (See T. C. Cain, D. M. Lubman, and W. J. Weber, Jr., Rapid Commun. Mass Spectrom., Vol. 8, pp. 1026-1030 (1994)). In addition, two different groups have reported the identification of intact bacteria with MALDI-TOF-MS. (See R. D. Holland et al., Rapid Communications in Mass Spectrometry, Vol. 10, pp. 1227-1232 (1996) and M. A. Claydon et al., Nature Biotechnology, Vol. 14, pp. 1584-1586).
The MALDI-MS technique is based on the discovery in the late 1980s that desorption/ionization of large, nonvolatile molecules such as proteins can be effected when a sample of such molecules is irradiated after being codeposited with a large molar excess of an energy-absorbing "matrix" material, even though the molecule does not strongly absorb at the wavelength of the laser radiation. The abrupt energy absorption initiates a phase change in a microvolume of the absorbing sample from a solid to a gas while also inducing ionization of the sample molecules.
Detailed descriptions of the MALDI-TOF-MS technique and its applications may be found in review articles by E. J. Zaluzec et al. (Protein Expression and Purification, Vol. 6, pp. 109-123 (1995)) and D. J. Harvey (Journal of Chromatography A, Vol. 720, pp. 429-4446 (1996)), each of which is incorporated herein by reference. In brief, the matrix and analyte are mixed to produce a solution with a matrix:analyte molar ratio of approximately 10,000:1. A small volume of this solution, typically 0.5-2 μl, is applied to a stainless steel probe tip and allowed to dry. During the drying process the matrix codeposits from solution with the analyte.
Ionization of the analyte is effected by pulsed laser radiation focused onto the probe tip which is located in a short (.about.5 cm) source region containing an electric field. The ions formed at the probe tip are accelerated by the electric field toward a detector through a flight tube, which is a long (.about.1 m) field free drift region. Since all ions receive the same amount of energy, the time required for ions to travel the length of the flight tube is dependent on their mass. Thus, low-mass ions have a shorter time of flight (TOF) than heavier ions. All the ions that reach the detector as the result of a single laser pulse produce a transient TOF signal. Typically, ten to several hundred transient TOF mass spectra are averaged to improve ion counting statistics.
The mass of an unknown analyte is determined by comparing its experimentally determined TOF to TOF signals obtained with ions of known mass. The MALDI-TOF-MS technique is capable of determining the mass of proteins of between 1 and 40 kDa with a typical accuracy of ±0.1%, and a somewhat lower accuracy for proteins of molecular mass above 40 kDa.
It is an object of the present invention to provide an improved mass spectrometric method for identifying bacteria at the genus, species and strain levels.
Another object of the present invention is to provide such a method which may be applied to whole bacteria cells in the laboratory or in field situations.
Yet another object of the present invention is to provide a library of genus, species and strain specific biomarkers for use in bacteria identification by mass spectrometry.
These and other objects are satisfied by the present invention which includes a method for generating unique mass spectral profiles, as well as individual biomarkers, of the proteinaceous material in bacteria extracts or whole bacteria cells by MALDI-TOF-MS, and the specific biomarkers produced by the method. The method has been described in detail elsewhere. See T. Krishnamurthy, et al., Rapid Communications in Mass Spectrometry, Vol 10, pp. 883-888 (1996). T. Krishnamurthy and P. Ross, Rapid Communications in Mass Spectrometry, Vol 10, pp. 1992-96 (1996), and T. Krishnamurthy and P. Ross, Proceedings of Seventh National Symposium on Mass Spectrometry, Indian Society for Mass Spectrometry, pp. 105-122 (November 1996), each of which is incorporated herein by reference.
A sample mixture is prepared by mixing a matrix solution with an unknown bacteria sample which comprises a protein extract or whole cells. A small aliquot 1 μl of the sample mixture is subjected to MS analysis in a MALDI-TOF instrument equipped with a nitrogen laser. Mass spectra are averaged over 100-200 individual laser shots, collected, and compared to MALDI-TOF-MS spectra of protein extracts or whole cells of known bacteria to identify the unknown bacteria sample.
As stated above, the present invention provides a method for generating unique mass spectral profiles for bacteria protein extracts or whole bacteria cells. These profiles contain proteinaceous biomarkers which distinguish between bacteria of different genera, species and strains. Comparable profiles are generated when the method is performed using different MALDI-TOF instruments from different manufactures. The invention will be further explained by reference to the nonlimiting examples set forth below.
Sample Preparation for MS Analysis
A number of pathogenic and nonpathogenic bacteria of different genera, species and strains were analyzed. The pathogenic bacteria strains were supplied as γ-irradiated and lyophilized samples by the U.S. Army Laboratories at Dugway Proving Ground, Utah. Nonpathogenic bacteria cells of different strains were grown in-house by incubating for 24 hrs. at 37° C. on trypticase soy agar or nutrient agar plates, harvested and lyophilized
Cell lysis
Lyophilized bacteria cells (1-2 mg) were suspended in 50 μl of a solution containing 10 mM Tris-HCl (pH 8.0), 0.1% sodium dodecyl sulfate (SDS), and 0.1 mM β-mercaptoethanol (BME). Alternatively, the same quantity of lyophilized bacterial cells were suspended in 50 μl of 10 mM Tris-HCl (pH 8.0), the suspended cells were sonicated using an ultramicro cell disruptor tip for 5 minutes, and then SDS and BME solutions were added to final concentrations of 0.1% and 0.1 mM, respectively.
The resulting cell solution, non-sonicated or sonicated, was incubated at 95° C. for 10 minutes, cooled, diluted with 50 μl of deionized water, and treated with 10 μl of a DNAase 1 solution (1 mg/ml). The reaction mixture was incubated at ambient temperature for 10 minutes and centrifuged. Spectrograde methanol (1 ml) was added to the supernatant liquid and the resulting solution was incubated at room temperature for 10 min. The solution was then centrifuged for 10 min. and the supernatant liquid decanted and discarded. The precipitate was air dried for 5-7 min and resuspended in 50-100 μl of a mixture of equal volumes of 100 mM ammonium bicarbonate and 1 mM calcium chloride. The resulting bacteria protein extracts were either used immediately for mass spectral analysis or were stored at -20° C. prior to use.
MALDI-TOF-MS Analysis
Matrix solutions contained about 10 mg/ml of either 4-hydroxy-α-cyano-cinnamic acid (4 CHCA; 10 mg/ml) or 3,5-dimethoxy-4-hydroxy cinnamic acid (sinapinic acid) in an aqueous solvent solution comprising 0.1% aqueous trifluoroacetic acid (TFA) and acetronitrile in a ratio of 70/30 (v/v).
Sample mixtures were prepared as follows. For analysis of bacterial protein extracts, 1 μl of a protein extract prepared as described in Example 1 was mixed with 9 μl of matrix solution. For analysis of whole cells, a small quantity (0.1-0.2 mg) of intact, whole cells are suspended were added to 20 μl of aqueous buffer, typically 0.1% trifluoroacetic acid, vortexed for 30 seconds, and 1 μl of the resulting suspension was either frozen for later use and thawed and combined with 9 μl of a matrix solution or used immediately.
The sample mixture, containing protein extract or whole cells, was vortexed for about 30 seconds and a 1 μl aliquot was then deposited onto the stainless steel autosampler pins of a Vestec 2000 MALDI-TOF-MS instrument (Vestec Instruments, Houston, Tex.) and allowed to air dry. The dried sample was irradiated at 337 nm from a nitrogen laser with the mass spectrometer operating in linear mode (reflection mode not operational). All experiments were conducted with the instrument operating at 26 kV acceleration voltage in linear mode, which gives an ion flight path of 1.25 meters. The instrument was also equipped with a low mass ion gate which substantially improves the sensitivity of the instrument to higher mass ions by rejecting ions below 1500 Daltons.
Mass spectra obtained during 100-200 individual laser shots, at a laser intensity approximately 10-30% above the threshold for ion appearance, were averaged and collected. The mass spectra were externally calibrated using calibrant mixtures comprising 1 pmol each of cytochrome c (12360.5 Da) or bovine insulin (5733.5 Da). Separate calibration files were generated for each sample carousel and for different days. Comparable data is achieved with internal calibration, when the concentration of the internal standard in the sample does not exceed 0.5 pmole/μl.
The mass spectra of protein extracts from mechanically and chemically lysed bacteria were found to be very similar for a particular bacterial strain. Ions of lower molecular weight were generally present in greater quantities, and thus more easily detected, in the extracts from sonicated bacteria, particularly when the bacterial sample was a gram-positive bacteria.
It was also observed that low ionization occurred when too much bacterial or protein extract was present in the sample. Thus, it is preferable to determine the optimal ratio of matrix to protein extract or whole cells for each bacteria sample. This may be done by serially diluting the protein extract or whole cell suspension in 0.1% TFA and mixing 1 μl of each dilution with 9 μl of matrix solution.
By comparing the mass spectra of samples from a number of bacteria, ions that are specific to the genus, species and/or strain of these bacteria were identified. Repeated experiments demonstrated that the measured mass/charge ratio (m/z) for a particular ion obtained with a protein extract or whole cell suspension varies slightly from experiment to experiment, within a range of 0.1%. This variation is due to the limitations observed in the mass assignments during MALDI-TOF-MS analysis. The average of the m/z ratios for that ion measured in 4-5 assays is the biomarker that was assigned to that bacteria. Any m/z measurement falling within ±0.1% of this biomarker is considered to be a genus, species or strain biomarker for that bacteria.
Mass spectral analysis of protein extracts according to the above described process consistently produced genus, species and strain specific biomarkers as shown in Table 1 below. TABLE 1 Biomarkers for Bacteria Protein Extracts Genus Species Strain Biomarkers Biomarkers Biomarkers Organism (m/z) (m/z) (m/z) Bacillus anthracis, 6680, 6837 2385, 3991, 4505 vollum 4313 Bacillus anthracis, 6680, 6837 2385, 3991, 2789 sterne 4313 Bacillus anthracis, 6680, 6837 2385, 3991, 2850 zimbabwe 4313 Bacillus thuringiensis, 6680, 6837 3932 5916 4A1 Bacillus thuringiensis, 6680, 6837 3932 4871, 7845 4A2 Bacillus thuringiensis, 6680, 6837 3932 2864, 4074, 4L2 4548, 5781 Bacillus cereus, 6680, 6837 5269, 5537, N/A 6E1 7365, 9533 Bacillus subtilis, 6680a 3757, 3871, N/A 3A1 6304, 9146 Brucella melitensis, 7379, 8031, 9060 N/A N/A melitensis wild Brucella melitensis, 7354, 8030, 9071 N/A N/A REV-1 vaccine Brucella melitensis, 7378, 8020, 9080 N/A 5844 Suis wild Brucella melitensis, 7379, 8020, 9088 N/A N/A abortus wild Yersinia pestis, 2460, 3223, 3297, N/A 7335 India 4361, 4437 Yersinia pestis, 3223, 3297, 4361, N/A N/A La Paz 4437 Yersinia pestis, 2453, 3223, 3297, N/A 5196, 5638, Nairobi 4361, 4437 7599, 9637 Francisella tularensis, 2622, 2770, 3205, N/A N/A type B 3274, 4120, 4747 a Doubly charged ion, corrected mass/charge ratio N/A - Data not available
The data in Table 1 include biomarkers which distinguish between four species of Bacillus. B. cereus and B. thuringiensis have distinct species specific biomarkers at m/z 9533 and 9652, respectively. Similarly, the ion observed at m/z 4313 appears to be specific for B. anthracis.
Different strains of a bacteria species can also be distinguished by MALDI-TOF-MS analysis of protein extracts according to the invention. As shown in Table 1, biomarkers specific for representative strains of B. anthracis and B. thuringiensis were identified.
Similarly, mass spectral analysis of intact, whole cells by the above procedure also produced biomarkers which distinguished between bacteria at the genus, species and strain levels. Data for several bacteria strains are listed below in Table 2: TABLE 2 Biomarkers for Whole Bacteria Cells Genus Species Strain Biomarkers Biomarkers Biomarkers Organism (m/z) (m/z) (m/z) Bacillus anthracis, 4339, 5178, 6436, 5422, 7780, 2880 zimbabwe 6686, 7377, 9640 9982 Bacillus thuringiensis, 4335, 5174, 6440, 3253, 5353 4D5 7378, 9640 Bacillus cereus, 4334, 5165, 6437, 3015, 4816 6E1 6680, 9640 Bacillus subtilis, 4340 6913, 9130 YB 886 Brucella melitensis, 7403, 9087, 2700, 10287 REV-1 vaccine 16100 Brucella melitensis, 7402, 9083, 10280 4512, 7664 Abortus 2308 16082 Yersinia pestis, 5946, 7288, 9167, 195P India 10881 Francisella tularensis, 4725, 6725, 9447, type B 10280 Blank boxes mean Not tested.
The above data demonstrate that MALDI-TOF-MS analysis of whole cells is capable of distinguishing between virulent and nonvirulent strains of bacteria. For example, REV-1 strain of Brucella melitensis is a non-pathogenic strain which is commonly used as a vaccine for humans and livestock, whereas the Abortus strain of B. melitensis is very pathogenic. The similar ionic profiles observed during analysis of whole cells from these two strains are easily distinguished by the presence of ions at m/z 4512 and 7664 in the Abortus strain which are lacking in the REV-1 strain.
Comparison of the biomarkers generated from protein extracts and whole cells showed a number of biomarkers in common and additional, unique biomarkers for the intact cells. The data for a number of representative bacteria strains are shown in Table 3 below: TABLE 3 Comparison of Biomarkers for Protein Extracts and Whole Cells Common Unique whole cell Biomarkers biomarkers Organism (m/z) (m/z) Bacillus anthracis, Sterne 4340, 6684, 6832 2580, 5171, 6431, 7374, 7772, 9640, 9981 Bacillus anthracis, 4342, 6685, 6828 2880, 5178, 6436, 7377, Zimbabwe 7780, 9988 Bacillus anthracis, Vollum 4339, 6683, 6837 2580, 5172, 6428, 7379, 7780, 9988 Bacillus cereus, 6E1 2470, 6682, 6827 3015, 4816, 5174, 6437, 9634 Bacillus thuringiensis, 4D5 4335, 6685 5174, 6440, 7378, 9640 Bacillus subtilis, 3898, 9130 4340, 6913 Brucella melitensis, REV-1 6650, 7044, 7341, 2700, 7403, 9087, 9071 10287, 16100 Yersinia pestis, 195P India N/F 5946, 7288, 9167, 10881 Francisella tularensis, N/F 4725, 6725, 9447, 10280 N/F - none found
Effect of Growth Conditions on Biomarkers Generated by MALDI-TOF-MS
The potential use of pathogenic bacteria by terrorists or by a country at war is a major international concern. Thus, it would be very useful if the source of any release could be identified. Intelligence information indicates that various countries, terrorist organizations, etc. may culture pathogenic bacteria under particular growing conditions. The present invention is capable of distinguishing between the same strain of bacteria grown under different conditions.
For example, mass spectra of whole Francisella tularensis cells grown in four different media (ISOVitalex Agar, BCYE Agar, Mueller-Hinton Broth, and Mueller-Hinton Agar) were very similar to each other and the identification of the strain could be determined using previously assigned biomarkers (data not shown). However, comparison of the spectra showed the presence of distinct ions (see Fig 10 of T. Krishnamurthy and P. Ross, Proceedings of Seventh National Symposium on Mass Spectrometry, Indian Society for Mass Spectrometry, pp. 105-122 (November 1996) for each growth medium.
Similarly, MALDI-TOF-MS analysis of whole cells according to the invention is capable of identifying a bacterium regardless of whether it is in a vegetative or sporulated form. These form-specific biomarkers can be applied for determining the growth conditions which may help identify the source of a bacteria sample collected from a release site.
For practical utility, mass spectral analysis of bacteria samples should be robust, i.e., not affected by various contaminants that may be present in the sample. This need is met by the present invention.
For example, the biomarkers generated by MALDI-TOF-MS of whole cells according to the present invention are not significantly affected by the pH 3-9 of the buffer used for bacteria suspensions.
In addition, unknown bacteria samples mixed with soil have been identified using previously assigned biomarkers.
Analysis of a Pathogenic Bacteria Mixture
Whole bacteria cells of four pathogenic bacteria, Bacillus anthracis, Yersinia pestis, Francisella tularensis, and Brucella melitensis, were mixed in various possible combinations and subjected to MALDI-MS-TOF analysis according to the method followed in Example 2. The mass spectra obtained with each mixture (data not shown) were compared with predetermined biomarkers for these organisms. The results demonstrated that each pathogen could be detected and identified in the presence of any of the either three pathogens. Moreover, analysis of a mixture of all four pathogens showed that the each of the four bacteria in the mixture could be detected and distinguished based on the observed biomarkers in the MALDI-MS spectrum of the mixture.
While the foregoing invention has been described in detail by way of illustration and example for purposes of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.
<- Previous Patent (Process for the manu...) | Next Patent (Acetyl-CoA carboxyla...) ->


Greetings everybody! Hope all is well out there. As I write this column, we find ourselves in the third week of some wonderfully warm and spring-like weather. It's been great. It's hard to believe that we're already having to put a "3" in the date section of our checks, isn't it? And as March comes, so does St. Patrick's Day—one of my favorite celebrations.
The old saying is that everybody's Irish on St. Patrick's Day. I'm actually part Irish so I have somewhat of a vested interest in this celebration. As far as I've been able to gather, my great, great, great, great-grandfather, James McCart, was the one who traveled from the Mother Country to the States. It was his son and my 3rd great-grandfather, William Marion McCart Sr., who brought his clan to the Covington area in the mid 1800's by way of Abbeville , SC and Lavonia , GA. But that is another tale for another time; one that I plan to write about in a future column.
So, what is St. Patrick's Day exactly? It's the celebration of St. Patrick, a 5th century A.D. Christian missionary who spread the good word and "cast out the serpents" from Ireland. The serpent bit is not literal or factual as it is believed that post-ice age Ireland never had any snakes. The serpents are more likely a symbol of the pre-Christian paganism and heathenism that gripped Ireland prior to Patrick's arrival. More specifically, the Druids were the primary religious force in Celtic Ireland during this time. The Druids also acted as judges, teachers, and historians. They were also, unfortunately, the ones who carried out the sacrifices.
Patrick, by all accounts, was a great man and almost single-handedly turned a pagan society into a Christian one. The celebration enjoys much interest in the cities and areas of our country that traditionally have had a lot of Irish immigrants. Boston , New York , Chicago , and other cities have large parades, green rivers, and lots of other festivities. Good ole Savannah , GA is known as one the best destination locations for those wanting to celebrate St. Patrick in the South. And as a quick aside, it's not St. Patty's Day—it's St. Patrick's Day!
But perhaps the best part of this holiday, for me personally, is that it is a perfect opportunity to fix one of my all-time favorite meals—Corned Beef, Potatoes, & Cabbage. I've been told by a few folks that I make the best they've ever had, so I thought I'd share my recipe with y'all.
Buy yourself a corned beef brisket at the store. It will come with some seasonings. Get yourself a bag of new potatoes, 2 medium onions, and a head of cabbage. Some say you have to use carrots to make it a truly Irish dish, but I disagree. I love carrots in a pot roast, but not for this.
I like to do mine in a Crock-Pot. Put it on high and fill it a little under halfway with water and add 6 oz. of Harp Lager and 2 shots of Irish Whiskey and throw in your brisket with the seasonings it came with. Then either quarter, third, or halve your potatoes (depending on how big they are) and throw them in. I'll usually use 10-12 smaller ones or 6-8 larger ones. Add some of salt and pepper and let it cook for about an hour. Then cut up your onion and put that in with a touch more whiskey and beer (about a shot of the former & approx. 4 oz of the latter. Turn it to low and let it cook during the day while you're at work. Then about an hour before you're ready to eat, cut up your cabbage and put it in and add a touch more whiskey, beer, salt, and pepper. An hour later, and you're ready to eat. You can also do it in the oven at 350. Do the brisket and potatoes for an hour; then the onions for an hour; and then the cabbage for an hour.
When you're ready to serve, remember to cut the meat against the grain. If you're industrious, you can Google the recipe for Irish Soda Bread to go with the meal. I just use loaf bread. In terms of drink, I like to accompany my meal with Bushmills, either neat or on the rocks, and a nice "Half & Half."—never to be confused with its more popular cousin, the "Black & Tan", a ½ & ½ is created by pouring a half glass of Harp Lager and topping it with an equal amount of Guinness Stout.
Happy St. Patrick's Day everybody! And remember to wear something green or you just might get pinched by a rogue Leprechaun!


A two-day South Asia Communication Conference (SACC), organized by the School of Mass Communication where delegates from India, Bangladesh and Nepal participated, was held at the Conference Hall-8 of KIIT Deemed to be University during December 21 and 22, 2018. The theme of the conference was 'Media and the Marginalised Communities'.
The inaugural function of the first edition of SACC was graced by Prof Biswajit Das, President, All India Communication and Media Association (AICMA), Prof Hrushikesha Mohanty, Vice-Chancellor, KIIT Deemed to be University, Dr Sasmita Samanta, Registrar, KIIT Deemed to be University, Prof Vibodh Parathsarthi, Associate Professor, Centre for Culture Media & Governance, Jamia Millia Islamia, Prof Himansu Sekhar Khatua, CEO of School of Mass Communication, School of Fashion Technology and School of Film and Media Sciences, Dr Bidu Bhusan Dash, Course Coordinator, KIIT School of Mass Communication and Convener of the conference. Conference organisers Ms Ruby Nanda, Faculty Associate, KIIT School of Mass Communication and Mr Sudarshan Sahoo, Faculty Associate, KIIT School of Mass Communication were also present.
Prof Biswajit Das delivered the inaugural address. He appreciated the commitment of Dr Achyuta Samanta, honourable founder of KIIT and KISS for providing quality education. Prof Das urged in his inaugural address, "There is a need for documentation and materials related to other countries in South Asia. There is a lack of organized platform for understanding South Asia beyond political and defence strategies". Congratulating the School of Mass Communication for the initiative, he expressed his hope that KIIT Deemed to be University will take a lead in this area.
In the same line, Prof Hrushikesha Mohanty, Vice-Chancellor of KIIT Deemed to be University said that there is a need for a repository of knowledge of South Asia. Congratulating the School of Mass Communication for its effort in organizing the conference, he said that this will also be a great learning experience for the young faculty members of the school. He called for the South Asia Communication Conference to be an annual event of the school.
Extending a warm welcome to the delegates from across the South Asian countries, Dr Sasmita Samanta, Registrar, KIIT Deemed to be University, talked about her own journey in the university. Speaking about the spectacular heights that KIIT has touched in its journey during the short span of 20 years, she said, "Everyday in KIIT, we come here thinking that history will be created today." Besides, Dr Samanta also conveyed that KIIT Deemed to be University is in race for the 'Institutions of Eminence' tag by the Central Government.
Eminent media academician Prof Vibodh Parathsarthi, Associate Professor, Centre for Culture Media & Governance, Jamia Millia Islamia, New Delhi, the keynote speaker of the conference, talked about how India played a dominant role in the knowledge platform in South Asia. He spoke on 'Anxiety amidst Abundance: A Plea for Media Diversity' in his key note address. He stressed on the need for further study on media diversity and made several deliberations to support his views.
Delegates from India, Bangladesh and Nepal were present in the Conference. As many as 21 researchers from the three countries presented their research findings and discussions in six thematic sessions spanning over two days.
On the valedictory session, Prof Biswajit Das suggested that KIIT should take the lead and start a South Asian Communication Centre. He went on describing that the centre should give emphasis on six parameters like networking, archiving, interdisciplinary dialogue, new methodology, fellowship and publication plans.
Subsequently, Prof H S Khatua, CEO of the school also accepted the proposal and assured to incorporate the suggestions as soon as possible. "KIIT School of Mass Communication will set up a Centre for South Asia Communication and will open a website within one year", announced Prof H S Khatua. He also shared the plan of publishing a research journal from the conference proceedings and sought suggestions from the delegates.
The guest of honour, Prof. Mohammad Sahid Ullah, Professor, University of Chittagong, Bangladesh, quoting Dr Achyuta Samanta, said that such a humble and great man is an inspiration for all.
Meanwhile Prof Mrinal Chatterjee, Centre Director, IIMC Dhenkanal mentioned the proposal of South Asia Communication Centre a welcome step.
AICMA was the knowledge partner of the Conference, while Kalinga TV was the media partner. The Conference was supported by NALCO.
Previously, the School of Mass Communication had organized the National Conference on Communication for Social Change, Development and Empowerment in Rural India on 4-5 December 2017 jointly by the KIIT School of Rural Management (KSRM) and School of Mass Communication, KIIT University, Bhubaneswar and supported by UNICEF India.


A Macy's in Century City, Calif.
You aren't imaging things if you think a ton of department stores have shuttered in the last three years.
According to a research report RBC Capital Markets released on Wednesday, Kohl's, Macy's (which also owns the upscale Bloomingdale's chain), J.C. Penney, Dillard's, and Sears (which also owns Kmart) have closed a total of 700 stores since 2013. That's almost 20% of those chains' store count.
The closings are meant to shed poor locations; compete with Amazon; and help companies manage the migration of their own business online.
Macy's, which is grappling with its worst slump since the Great Recession, announced last month it would close 100 more stores after the holiday season to make way for incoming CEO Jeff Gennette. When he takes the reins in February, Gennette will try to repair the iconic, diminished chain. The company, which is the result of the amalgamation of many regional chains in 2005, has struggled with its fair share of decrepit stores—but it chose to close locations in one fell swoop as opposed to just a few stores at a time.
In Kohl's case, the drop has been modest: the retailer closed 18 of its 1,150+ plus stores. But with its comparable sales on the decline after a brief turnaround, it wouldn't be surprising to see the company shutter more locations if it doesn't enjoy a decent holiday season.
Sears and Kmart have been closing dozens of stores as sales continue to plummet despite CEO (and top shareholder) Eddie Lampert's promises to reinvent the retailers.
It's not just the drop in mall traffic: the most recent earnings season showed top department stores, staples of American shopping for more than a century, are having a hard time holding their own against the likes of Amazon.com (amzn). And analysts expect those problems to persist.
In April, leading real estate analysis firm Green Street Advisors said the top department stores would need to close some 800 stores for sales-per-square-foot to get back to 2006 levels.
Still, wholesale closings present their own risks. As Ellison told Fortune this winter, stores play crucial roles in his company's e-commerce growth, allowing not them not only to compete with Amazon but also to serve as additional distribution centers and pick-up spots for online orders. Penney has no plans for any large-scale shutterings because the vast majority of its locations are profitable anyway, he added.
Yet it's pointless to operate stores no one goes to anymore or that are such dumps they hurt the overall brand for the retailer. If department stores struggle yet again this holiday season, expect a spate of store closing announcements the first week of January.


SINGAPORE - Eight decades of history at Sungei Road flea market will come to an end this July (2017).
In a multi-agency statement on Tuesday, the Government said that the hawking zone's last day of operation will be July 10. The flea market will make way for future residential developments..
The Straits Times understands that roads in the area could be re-aligned as well.
The statement was issued by the National Environment Agency (NEA), Ministry of National Development, Ministry of Social and Family Development, Workforce Singapore, National Heritage Board (NHB) and Singapore Police Force. It noted that the 31 rag-and-bone men from the Sungei Road site were not included in the Government's street hawker resettlement programme to purpose-built markets and hawker centres back in the 1970s and 1980s "because of their chosen trade".
They were given permits and allowed to continue at Sungei Road. Only 11 permit holders still operate there today.
The NEA will be offering these remaining permit holders the option of operating at lock-up stalls at selected hawker centres at subsidised rental rates following the closure of the flea market.
Meanwhile, vendors who are registered with the police under the Secondhand Goods Dealers Act will need to provide a new business address if they wish to continue to ply their secondhand goods trade elsewhere.
The authorities said that Social Service Offices will facilitate financial assistance and Workforce Singapore will provide employment services under existing schemes to eligible vendors who may require such help.
They added that while the site has had a long history, and holds special memories for many Singaporeans, "over time, the nature of the site has changed, as reflected in both the profile of vendors and buyers, and type of goods sold".
Previous articles have reported that "opportunistic traders" are attracted to the site, also known as Jalan Besar market, because of its rent-free and city location. The authorities have had to conduct checks on the sale of prohibited goods regularly.
Residents have also complained about the market located at the junction of Jalan Besar and Rochor Canal Road. One grouse is about traders who store goods in their void decks.
The Government said that such street trades should be allowed only to continue in designated venues such as trade fairs and flea markets, rather than on a permanent basis.
It noted that the NHB has conducted research and documentation efforts on the market and its vendors to preserve memories of the site.
Mr Koh Ah Koon, 76, the president of the Association for the Recycling of Second Hand Goods representing about 70 of about 200 stalls at the market, said he is disappointed by the news.
He told ST that the closure of the site could push some peddlers back to the ways of the past when they squatted illegally. He added that his association had proposed four alternative sites but these had been rejected by the authorities.
He said: "At least 80 per cent of us are elderly folk in our 60s, 70s and 80s who depend on our stalls for income. We hope we will be able to keep this traditional trade and way of displaying our wares alive."
Singapore Heritage Society president Chua Ai Lin said that it is a pity that alternative solutions were not found. She said the market is a good example of a functioning community and economic system and a facet of Singapore's rich and diverse urban life.
Dr Chua said: "This is the only free hawking zone in Singapore. We will be losing the sense of an organically formed flea market. A whole community will be dispersed and can no longer congregate as second-hand sellers.
"The Sungei Road flea market is an important part of our landscape that makes Singapore more than just shopping malls."
The flea market began as a small trading spot that sprouted along the river during the mid 1930s.
In 2011, the Sungei Road site was reduced to half its size in order to make way for the construction of the new Sungei Road MRT station.
The space allotted for each peddler is limited to a metre-by-metre and offered on a first-come-first-serve basis. Vendors are permitted to sell only second-hand or used merchandise.
The market is open from 1pm to 7pm daily and is popular with tourists, foreign workers and locals flock there on weekends to buy bits and bobs such as old coins, stamps, cassettes, jewellery, clothes and electronic gear.


America's top 20 inbound tourism markets includes a surprising mix, but as expected, overall rapid growth from Asia and Latin America points to where U.S. tourism efforts should be focused for the at least the next decade.
The number of Europeans visiting the U.S. continued to drop in 2012, while the number of Asian and Latin Americans quickly increased.
Data released by the U.S. Department of Commerce today details America's top inbound tourism markets from 2012.
Total visitor numbers increased 7 percent to a new record of 67 million international visitors. Arrivals were led by Canada and Mexico, which brought in 22.7 million and 14.5 million travelers a piece.
Of the top 20 inbound markets, all six that dropped in visitation were in Europe: UK (-1.9%), France (-3.2%), Italy (-6.8%), Spain (-13.3%), the Netherlands (-1.5%), and Ireland (-4.3%).
All other markets increased visitation with the greatest growth coming from Asia and Latin America. China posted a remarkable 35.5 percent increase in visitation followed by growth from Colombia (+21.2%), Venezuela (+20.3%), Argentina (+20%), and Brazil (+18.8%).


text:
MUS 113
2 in 1945 cherokee became ko ko recorded for savoy
School Binghamton University
Course Title MUS 113
Uploaded By horsecrazy7yay
this piece that amazed everyone who heard it. 2. In 1945, "Cherokee" became "Ko-Ko," recorded for Savoy Records. The owner of the label would not tolerate a copyrighted melody, so they left the "Cherokee" melody out. The pianist was supposed to be Bud Powel but he didn't show, so for this piece it is probably Gillespie on piano except when he is playing trumpet. The other pianist was Argonne Thornton (later Sadik Hakim). 10. "Embraceable You" 1. This Gershwin piece was recorded in 1948, but Parker avoids the melody. Instead he plays a popular 1939 melody, "A Table in the Corner," recorded by Artie Shaw. After Parker's impressive solo, a young Miles Davis takes the next solo. 11. "Now's the Time" 1. Parker considered bebop to be a collision of New York progressive music with Midwest blues. Blues during the 1940s had many faces, including swing band blues and Mississippi delta blues electrified by the likes of Muddy Waters. Parker's contribution was to add the chromatic harmonies of modern jazz and a fluid sense of rhythm to the vocal nuances of blues. 2. This piece is a Parker composition built on one riff. It was used later for a rock and roll hit called "The Hucklebuck" and was covered by many pop musicians. Parker didn't earn a cent from the royalties because the owner of Savoy Records owned the copyright. 3. This recording was made for a major label (Verve) so the sound is much better than in other Parker recordings. Nuances are more easily heard. 12. Bird's Last Flight 1. The Parker-Gillespie partnership ended in 1946 when the band went to Los Angeles and met with an indifferent response. Gillespie took the band back to New York, but Bird cashed in his ticket to get money for his heroin habit. He stayed in California for a year, taking drugs, and when the heroin ran out he turned to alcohol and barbiturates. 2. In 1947 Parker made some recordings for Dial, which showed his playing at its worst. Later that night he was found in his hotel lobby wearing only his socks. He was arrested and committed to the state hospital for six months.
3. Free from drugs, he returned to New York only to return to his habit. In the view of bandmate Miles Davis, Parker became "some kind of . . . clown." 4. With the help of Norman Granz, Parker found some commercial success with Mercury Records, where he recorded with strings. Just Friends from this session became his most successful record. But his drug addiction made him unreliable and wore him down. In 1955, he died. He was thirty-four; the coroner estimated his age as fifty-three. 13. The Elder Statesman 1. Gillespie disdained drugs and showed how bebop could act as a foundation for the professional jazz musician. 2. Upon returning from California in 1946, he started a big band using bebop arrangements. When not playing trumpet, he took his cue from his former boss, Cab Calloway, who balanced art with wit and silliness-a mix that could broaden the audience for bop.
PROFESSOR MarcusLalli
TAGS Music, On the Road
Learn more about On the Road with Course Hero's FREE study guides and infographics!
Most Popular Documents for MUS 113
3 Benny Carter recommended her to Chick Webb He became her legal guardian and
MUS 113 - Fall 2015
chapter 9 notes
1877 born in New Orleans 1895 started working in parades and other functions
Tin Pan Alley This refers to the popular music written between the 1890s and
2 He was born into a wealthy New York family He was attracted to the black
2 He usually played second trumpet there were a few exceptions such as
Ragtime Like jazz ragtime embodied the mix of African American and white art
Interested in Chapter 11
Bookmark Chapter 11.
He continued to explore music and became an elder statesman of jazz He died in
MUS 113 - Spring 2014
Chapter 11 Notes (Jazz in American Music)
5 Although he could play well while on heroin his constant movement from band
Chapter 11.docx
Parker stayed in New York for a year or so before going back to Kansas City for
MUH 3016 - Summer 2012
MUH 3016 Session 5-6 Lecture/Notes
In 1947 Parker made some recordings for Dial which showed his playing at its
MUSIC 3UU3 - Spring 2011
Jazz_PPT_ch11 (1)
o d J J Johnson on trombone Serge Chaloff and Leo Parker on baritone saxophone
MUSC 1236 - Spring 2013
Chapter 11 Notes.docx
solos and contributed new tunes to the group Joe Zawinul made an accordion when
Jazz History Final Study Guide

redpajama_set_name: RedPajamaCommonCrawl


What are your beliefs? Have you really thought about where they come from? I have always said if you believe something to be true then it is. If you believe you are going to have a "crappy" day, guess what…it will be! If you believe you will have an amazing time, chances are you will. What we believe we project.
As I work on my "vision" board of what I want to visualize for this new year, the more I think about it the more I see things that I want to bring to my reality. There really is no "secret" to success, it is really about manifesting what you want for your life.
As you start this new year, what would you like for you and for your family? Start right now by writing down what you would like. Then picture yourself enjoying what it is you would like to have. See yourself there.
This really and truly works, last year I was truly blessed with so many wonderful things that I imagined and received.
Let's bring this back to writing as that is what I teach. Would you love to write your story down this year? Do you believe you can? Most of our blocks in writing comes because of our belief that we are too busy to spend the time writing. This year let's take down those blocks of negativity and focus on what you can do.


CanadaVet.com Blog: Take Action Early To Avoid Flea Plagues!
As the temperature begins to slowly warm back up it is important for all pet owners to make sure that they are using flea control. Pet owners who don't take action will find they are simply setting themselves up for a flea plague.
Fleas and their eggs can survive indoors throughout the winter especially in a heated home as well as in the carpets, under furniture and in the pet's bedding, etc. It is these fleas that are laying the eggs in the environment. As soon as the weather warms up, these eggs hatch all at the same time, causing a flea problem in plague proportions.
To avoid this happening to your pets, act early and get one step ahead of any flea problem that may arise. You will be rewarded by happier and healthier pets and not have the need to battle a massive flea plague all summer!


Customer service is our top priority!. Now in its second edition, Lippincott Illustrated Reviews: Cell and Molecular Biology continues to provide a highly visual presentation of essential cell and molecular biology, focusing on topics related to human health and disease. Connecting readers with great books since 1972. May contain highlighting, handwriting or underlining through out the book. The book can be used as a review text for a stand-alone cell biology course in medical, health professions, and upper-level undergraduate programs, or in conjunction with Lippincott's Illustrated Reviews: Biochemistry for integrated courses. About this Item: Condition: Acceptable. Look for other fine titles in the Lippincott Illustrated Reviews series! A copy that has been read, but remains in clean condition.
About this Item: Condition: Good. The spine may show signs of wear. Connecting readers with great books since 1972. Look for other fine titles in the Lippincott® Illustrated Reviews series! Book is in Used-Good condition. About this Title Efficiently master essential cell and molecular biology information! This new addition to the internationally best-selling Lippincott's Illustrated Reviews Series includes all the popular features of the series: an abundance of full-color annotated illustrations, expanded outline format, chapter summaries, review questions, and case studies that link basic science to real-life clinical situations.
Access codes may or may not work. At ThriftBooks, our motto is: Read More, Spend Less. Customer service is our top priority. We may ship the books from Asian regions for inventory purpose. Territorial restrictions may be printed on the book.
Customer service is our top priority!. Sequential images detail how drugs work in dynamic detail, and enhanced review questions with answers equip you for success on your exams and beyond. Now in its second edition, Lippincott Illustrated Reviews: Cell and Molecular Biology continues to provide a highly visual presentation of essential cell and molecular biology, focusing on topics related to human health and disease. Ships same day or next business day! Now in its second edition, Lippincott Illustrated Reviews: Cell and Molecular Biology continues to provide a highly visual presentation of essential cell and molecular biology, focusing on topics related to human health and disease. We may ship the books from Asian regions for inventory purpose. This concise resource features clear, effective writing and hundreds of illustrations that break down complex information for rapid review.
This new addition to the internationally best-selling Lippincott's Illustrated Reviews Series includes all the popular features of the series: an abundance of full-color annotated illustrations, expanded outline format, chapter summaries, review questions, and case studies that link basic science to real-life clinical situations. Pages and cover are clean and intact. . This is a paper back book: This item is fairly worn but is functional. End Chapter Exercises may differ. Kindly provide day time phone number in order to ensure smooth delivery.
About the e-Book Cell and Molecular Biology Lippincott Illustrated Reviews Series pdf Lippincott's Illustrated Reviews: Cell and Molecular Biology offers a highly visual presentation of essential cell and molecular biology, focusing on topics related to human health and disease. May include supplemental or companion materials if applicable. Lippincott's Illustrated Reviews: Cell and Molecular Biology offers a highly visual presentation of essential cell and molecular biology, focusing on topics related to human health and disease. Lippincott® Illustrated Reviews: Pharmacology, Seventh Edition presents current drug information in an accessible format ideal for a fast, effective refresher on the essentials of medical pharmacology. All pages are intact, and the cover is intact. All pages are intact, and the cover is intact. It offers all the most popular features of the bestselling Lippincott Illustrated Reviews series, including abundant full-color, annotated illustrations, chapter overviews, an expanded outline format, chapter summaries, and review questions that link basic science to real-life clinical situations.
This book has many years of enjoyment left in the pages. At ThriftBooks, our motto is: Read More, Spend Less. Efficiently master essential cell and molecular biology information! The spine may show signs of wear. In case of orders from Europe, custom charges may comply by the relevant government authority and we are not liable for it. May show signs of minor shelf wear and contain limited notes and highlighting. A companion Website features the fully searchable online text, an interactive Question Bank for students, and an Image Bank for instructors to create PowerPoint® presentations. A companion Website features the fully searchable online text, an interactive Question Bank for students, and an Image Bank for instructors to create PowerPoint R presentations.
We ship Monday-Saturday and respond to inquries within 24 hours. Pages can include limited notes and highlighting, and the copy can include previous owner inscriptions. Connecting readers with great books since 1972. Legal to use despite any disclaimer. This new addition to the internationally best-selling Lippincott's Illustrated Reviews Series includes all the popular features of the series: an abundance of full-color annotated illustrations, expanded outline format, chapter summaries, review questions, and case studies that link basic science to real-life clinical situations.


For more info go to www.kibbletowerpetfeeder.com Kibble Tower pet feeder, cat & dog feeder. Challenge your pet with an interactive feeder. Slow down your pet's eating habits and have him work a little for their food. No need for batteries. Whether you free feed or not there is no change to your current habits. Both cats and dogs will easily pick it up. Crafted out of pine and finished with an all natural varnish it is sleek, aesthetic and fits in any room. Please leave a comment.
For more info go to www.kibbletowerpetfeeder.com Kibble Tower pet feeder, cat & dog feeder. Challenge your pet with an interactive feeder. Slow down your pet's eating habits and have him work a little for their food. No need for batteries. Whether you free feed or not there is no change to your current habits. Both cats and dogs will easily pick it up. Crafted out of pine and finished with an all natural varnish. It is sleek, aesthetic and fits in any room. Please leave your comments.


TeenHelp Mind and Body Depression and Suicide Triggering (Grieving): Need it to go Away!
This thread has been labeled as triggering, particularly on the subject of death or grieving, by the original poster or by a Moderator. The contents of this thread might therefore not be suitable for certain sensitive users. Please take this into consideration before continuing to read.
I don't know how much more I can hold onto for! Having a family member and a pet pass away both in two weeks. I hurt so much. I'm trying to be okay and happy and nothing is helping me to be okay and happy. I feel so depressed and hurt and part of me just wants to give up. What else can happen to us? I can't make this hurt and pain go away. What's the point now? Horrible things are always going to happen what's the point now. I can't think, this pain is not leaving me. What's wrong with me?
When we lose someone or a pet we go through grieve and many emotions. Sometimes we need to talk to a therapist about how we are grieving and how it's affecting us. Is this something you could ask for? To see if you can talk to someone about how you are feeling?
When we lose someone and a pet it can be hard to cope with or understand. Sometimes knowing why it happened can help us grieve or it may not help.
They ar both in your hearts. You have memories of them. Hold onto those memories. Choose memories that are warm to your heart and feel how they make you feel. You may feel upset because you miss them but also laugh at the moments you had with them that are good.
It will take time to grieve and it does hurt but it does get better over time.
If you need anything, feel free to reach out.
Nothing is wrong with you. The fact is that you are grieving and it can be quite a painful thing to deal with. I know things seem impossible right now but as time goes on you might find that the pain decreases. It can take a while for that to happen but please keep hanging in there.
I am curious if you've been able to talk to your parents or a counselor about how you are feeling? I know you might not have a strong desire to open up about these feelings but it could prove to be beneficial. If you opened up to your parents it's possible that they would want to try and support you through this.
There is nothing wrong with you at all. You are grieving. I am so sorry for your losses. I know how much this can hurt. I've watched my mom and grandma grieve over my great grandma it was really tough for them.
I know what you mean by bad stuff is always going to happen so you just want to give up now instead of deal with the bad stuff when it comes. I have felt that way so much. My suggestion is try not to worry about when bad stuff is going to happen and just focus on the here and now.
The people and pet that you have lost will always be in your heart, and I believe they are looking down at you and your family.
I don't know if you are up to doing this but it might be a good idea. You could look into grief therapy groups for teens and if you can find one by you, you might want to go and see what it is like. If it helps you then you can always go back.
If you ever need to talk I am always here.
Thank you so much for you're replies. I will try to find someone to talk to about this.
I'm here too and I know what it's like to lose someone dear.. I want you to know that you deserve happiness and you deserve to do whatever it takes to regain your happiness..


text:
Charles Massin
Les Carnets du Patrimoine - Haute-Provence et Vaucluse, 2000
Heritage of the French regions of Haute-Provence and Vaucluse.
Les Carnets du Patrimoine - Provence Côte d'Azur, 2000
Heritage of the French region of Provence Côte d'Azur .
Les Carnets du Patrimoine - Touraine et Sologne, 2000
Heritage of the French regions of Touraine and Sologne.
Les Carnets du Patrimoine  Anjou, 1999
Heritage of the French region of Anjou.
Copyright © 2002-2010 Urban-Resources All rights reserved Powered by PocketTV ©


text:
The Best Way To Say Goodbye To Your Year: My 2016 Anthems
By Chrissy Stockton, December 31st 2016
http://tcat.tc/2imt2sk
The best way to organize your thoughts about a year is to pick a song every month that represents its vibe in some way. Maybe the lyrics speak to a situation you were working on overcoming, maybe it played during a particular high or low point, or maybe it was just your most played song of that month. At the end of the year you have 12 songs, each one particularly powerful in transporting you to a particular moment and evoking the overall feelings that add up to your year.
I've been doing this for 10 years now and when I play my 2006 playlist, I can still remember why I felt like partying in September 2006 and why I broke up with a guy in February of that year and how the year felt as a whole.
To better understand how to make an annual vibes playlist, here's my 2016:
January — Only Girl In The World (Rihanna)
Want you to make me feel
Like I'm the only girl in the world
Like I'm the only one that you'll ever love
Like I'm the only one who knows your heart
Only girl in the world
Like I'm the only one that's in command
'Cause I'm the only one who understands
How to make you feel like a man
My friend made fun of me when we traded Spotify year in review playlists and he saw this old Rihanna song charted so high on mine. But it's a banger! And more importantly it clearly articulates the standards I have for my life. All I want is for all the people I respect and care about to make me feel like I'm the only girl in the world. It's really not that complicated.
I play this song whenever I do this thing I call instilling the low key fear of god in someone. Love is really important to me and I care a lot about making sure the people I care about in all forms of romantic/platonic/work relationships know how special they are. But this makes it easy for people to get too comfortable and so every once in awhile I have to remind someone that as sensitive and emotional as I am, I'm a rationality-driven person and if the hedonic calculus doesn't add up for me anymore, I'm cool making difficult decisions in order to make myself happier.
So I revisit the gold standard of having standards: make me feel like the only girl in the world. That's all.
February — Don't Fucking Tell Me What To Do (Robyn)
Your nagging is killing me
Ease up, you're killing me
Let go, you're killing me
Calm down, you're killing me
My god, you're killing me
If you cut open my heart, there's a Robyn song playing in there. She's so powerful and feminine and non-conventional. This song is about being a creator and a lover and working yourself to the bone for all the things you care about. And then, somehow, there's still outside pressure there when you're fully capable of putting enough pressure on yourself.
This was a month of taking space from other people's annoying opinions about what I'm supposed to be doing for them.
March — Wide Awake (Katy Perry)
I'm wide awake
Yeah, I was in the dark
I was falling hard
With an open heart
How did I read the stars so wrong
The beginning of the year was clearly full of angst for me. This is a song about letting people know you're not falling for what they are telling you. I don't really care about calling people out on their shit, but I do start to care when they think I don't realize what they are doing.
It also has a lot of philosophical connotations, which I like.
April — Heartbeats (Royal Teeth)
One night to be confused
One night to speed up truth
We had a promise made
Four hands and then away
Both under influence
We had divine scent
To know what to say
Mind is a razor blade
This is a cover of one of my favorite songs of all time (Heartbeats — The Knife, its v v v incredible). It's about sleeping someone when you're still in love with someone else and also wondering if you believe in god.
This is an especially beautiful cover I discovered this month that helped me fall in love with the song all over again.
I was seeing someone and battling the feelings of how good and deep our physical chemistry could feel while also being just totally insanely removed from any kind of thinking or love or real value. I wasn't used to that. It was the embodiment of that Tolstoy quote about how delusional we are as a human race that beauty is related to goodness.
(NEVER @ ME ABOUT THE JOSE GONZALEZ COVER OF HEARTBEATS IT SUCKS AND IS BLASPHEMY EXCEPT FOR THIS COOL SONY BRAVIA COMMERCIAL MY FRIENDS DID).
May — Southern Cross (Crosby, Stills & Nash)
When you see the Southern Cross for the first time
You understand now why you came this way
'Cause the truth you might be runnin' from is so small
But it's as big as the promise, the promise of a comin' day
I was getting ready for a summer of travel and I was captivated by the idea of how this constellation of stars could seem like magic the first time you saw it.
June — Record Year (Eric Church)
Slowly plannin' my survival
In a three foot stack of vinyl
I had made the decision to move out of my apartment and to stay in Minnesota and to live in my own apartment for the first time ever. It was the beginning of summer which should be full of Britney jams but I was already nostalgic about how the year would feel because I was making such big decisions. I especially related to it as I packed up my stuff and sat in literal piles of Chrissy.
A close second here would be Wiz Khalifa's No Sleep because I remember being at a giant MTV beach house with all my friends in Santa Cruz and someone put this song on the Sonos and everyone died and devolved into dancing and talking about how this is probably the most underrated song of all time and it was a MOMENT.
July — YOUTH (Troye Sivan)
What if, what if we run away
What if, what if we left today
What if, we say goodbye to safe and sound
What if, what if we're hard to find
What if, what if we lost our minds
What if, we looked and fall behind and then never found
In July I fell in love with Troye Sivan and HOW COULD YOU NOT ???? The only thing I listened to the entire month of June is Troye Sivan. Watch the video and get with the program.
There was a day when my friend and I drove to a giant luxury pool in the suburbs to get served mojitos by poolboys all day and played Troye Sivan the entire there way and back.
August — Work From Home (Fifth Harmony)
And I don't need no explanation
'Cause baby, you're the boss at home
This is one of the best songs of our generation. And it's also perfect for me because I have a very good work ethic while also being totally into the concept of a guy who's 'the boss at home.' It's also impossible to listen to without a good visceral hair flip or two. I remembered it being on the radio every time I turned my rental car on in California and then in August every time I needed to turn up my focus and #grind I'd play this one hour loop of the song and get in the zone.
Also Real Chad Johnson is obsessed with Work From Home which is just the telos of all my guilty pleasures.
September — Acid Tongue (Jenny Lewis)
I went to a cobbler
to fix a hole in my shoe
he took one look at my face and said
"I can fix that hole in you."
The first time I heard this song I was obsessed with it and listened to it hundreds of times in a row. I instantly and viscerally related to the line about having a 'hole' in you. I drove through Minnesota and Iowa and Nebraska and Colorado this month. I spent a lot of time in my car and with my radio and with my heart and my mind.
I tried to get ready for this big trip by thinking about all the things I was going to think about on my drive and all the things I was going to leave behind in Minnesota. It didn't work. I didn't get to leave all the things and all the people I wanted to leave behind, but it was still cathartic in all the ways I wanted it to be cathartic.
October — Vindicated (Dashboard Confessional)
I am selfish
I am wrong
I am right, I swear I'm right
Swear I knew it all along and
I am flawed
But I am cleaning up so well
I am seeing in me now the things you swore you saw yourself
A holdover from my Colorado trip. I spent my long, straight drive through Nebraska playing A Mark, A Mission, A Brand, A Scar straight through, and then all my other Dashboard favorites. It was such a fun experience because I knew all of it so viscerally. These were lyrics I had screamed before. Suddenly they were relevant again and the giant catharsis was coming at the end of the road, at the end of the album, at the end of the trip. It was medicinal.
November — Lucky Now (Ryan Adams)
I don't remember were we wild and young
All that faded into memory
I feel like somebody I don't know
Are we really who we used to be
Am I really who I was
In November I was thinking about getting older and being at peace with it. I feel safe enough that I've accomplished enough to be 31. Ryan's one of my favorite artists, ever, and he's a decade older than me but this song really resonated with me this year. I used to be a very cool internet teen and then a 20-something with a viral tumblr who went to cool raves all the time and I'm so not cool anymore. Which I am happy with but is also a loss I am grieving. There is only forward.
December — Not The Doctor (Alanis Morisette)
I don't want to be the filler if the void is solely yours
I don't want to be your glass of single malt whiskey
Hidden in the bottom drawer
I don't want to be a bandage if the wound is not mine
Lend me some fresh air
I don't want to be adored for what I merely represent to you
I don't want to be your babysitter
You're a very big boy now
I don't want to be your mother
I didn't carry you in my womb for nine months
I'm ending the year like I began it, which is kind of a bad thing because the vibe is having to remind people that all relationships are an equation and if you don't get out of them what you put into them you need to let go and let them sink.
This reminds me that in 2017 I need to try harder to have a more positive beginning of the year because I think that influences the entire rest of your year. I don't want to start and end by having to prove myself when that's always been so self-obvious in the past. That's not something that should be complicated and I've learned this year that people who make it complicated are too bad at thinking and too much work to be worth it. People are worth it or they aren't, that's the easy part.
Do you see that this is the way to remember a year? Leave your Spotify playlist link in the comments.
2016 2017 Memories Music nostalgia Playlists Spotify
Scrapbooks Are Not Chic: How To Remember The Important Things About Each Year Through Music
The Best Sad Coffee Songs
The Prettiest Songs Of All Time
21 Terrible 90s Songs That Everyone Secretly Loves
A Ranking Of Every Song Off Of 'Reputation' By Taylor Swift
The 10 Worst Rap Lyrics Of All Time
A 160+ Cover Song Playlist


text:
Unidentified, Flask, glass, Smithsonian American Art Museum, Gift of John Gellatly, 1929.8.147.30
Most of the glass vessels in this case date from the first century BC to the fourth century AD. Early glass vessels were made in the Middle East and Egypt using the core-forming technique, in which molten glass was poured over a clay core and decorated with threads of colored glass. During the first century AD, Rome became the center of glassmaking, and the invention of blown glass led to new methods, including free-blown glass, which could be decorated by pinching, rolling, or dragging the surface, and mold-blown glass, in which the molten glass was blown into a terra-cotta mold. The Romans also developed stratified glass, in which different colored canes were fused together and blown [see 1929.8.147.1, 1929.8.147.2], and millefiori glass (Italian for "one-thousand flowers"), in which colored strips of glass were joined together into a rod, cut into slices, and fused into bowls and cups [see 1929.8.147.13, 1929.8.157.9]. Many glass vessels were buried in the tombs of wealthy Romans and this contact with damp soil over hundreds of years caused the surface of the glass to deteriorate and become iridescent [see 1929.8.147.37, 1929.8.157.22].
Gift of John Gellatly
#B6A37B
#FFFFFA
Plate with the Contest between the Muses and the Pierides
Orazio Fontana
Apauly-Tustennuggee
Charles Bird King
enamel and gilded metal
Unidentified (Roman)
Tutti Frutti Goblet (purple top; rose base)
Robert Dane


text:
Maurice Maréchal, né à Dijon le  et mort à Paris  le , est un violoncelliste français.

Biographie

Jeunesse et formation

Maurice Maréchal est né à Dijon le . Il est le fils de Jules Jacques Maréchal employé aux Postes et Télégraphes et de Marthe Justine Morier son épouse.

Après avoir étudié au conservatoire de sa ville natale, il entre en 1905 au Conservatoire de Paris où il remporte son premier prix de violoncelle en 1911.

Première Guerre mondiale

Trois ans plus tard, la France entre en guerre ; Maurice Maréchal était depuis  incorporé au 74e régiment d'infanterie de Rouen dans le cadre de la Loi des Trois ans. Âgé de 22 ans, il est mobilisé (matricule 4684, classe 12, soldat de , brancardier et agent de liaison cycliste) au 274e régiment d'infanterie. Il raconte son quotidien d'août 1914 à février 1919 dans ses carnets intimes. Quelques-unes de ses lettres sont édités dans le livre Paroles de Poilus.

Deux camarades de régiment, Antoine Neyen (1877-1915) et Albert Plicque (1885-1915), lui confectionnent entre mai et juin 1915 un violoncelle de fortune avec du bois de récupération. Maurice Maréchal le surnommera le « Poilu » et le jouera lors d'offices religieux ou pour des officiers entre juin 1915 et octobre 1916. Au sein de la  d'infanterie de Mangin, mélomane qui encourage la musique, il fait la connaissance d'autres musiciens comme Gustave Cloëz ou le pianiste Henri Magne et forme avec Lucien Durosoir au premier violon, Henri Lemoine au second violon et André Caplet à l'alto, un quatuor qui se produit devant l'état-major.

Il a obtenu la Croix de Guerre en 1916.

Carrière musicale

Après la guerre, Maurice Maréchal intègre en 1919 les Concerts Lamoureux pour un an. On le retrouve en 1926 dans l'Orchestre de New York. Il entame alors une carrière de soliste qui le mène sur tous les continents. Son ami Émile Poillot l'accompagne au piano lors de plusieurs tournées en Espagne (1925 et 1926), en France (1928), à Singapour (lundi ) et aux Indes néerlandaises (août et ).

En 1942, il est nommé professeur au Conservatoire de Paris, poste qu'il quitte un an avant sa mort qui survient en 1964, alors qu'il est âgé de 72 ans. Parmi ses élèves figurent Christine Walevska, Alain Lambert, Jean Deplace et Alain Meunier.

Il a créé notamment la Sonate pour violon et violoncelle de Maurice Ravel, l'Épiphanie d'André Caplet, les concertos d'Arthur Honegger et de Darius Milhaud.

Ses neuf carnets intimes, rassemblés avec des lettres de Lucien Durosoir, sont parus en 2005 dans un ouvrage intitulé Deux musiciens dans la Grande Guerre. Son élève Alain Lambert lui a consacré un livre, Maurice Maréchal, la voix du violoncelle.

Maurice Maréchal est décédé le  à son domicile 6, rue Freycinet à Paris  des suites d'une opération du rein. Ses obsèques ont eu lieu en la cathédrale Saint-Bénigne de Dijon le  ; il est inhumé au cimetière des Péjoces à Dijon.

Décorations
Maurice Maréchal était officier de la Légion d'Honneur et titulaire de la Croix de Guerre.

Bibliographie
Son épouse, Loïs Perkins-Maréchal, évoque longuement leur vie commune et l'activité artistique de son époux dans L'Amérique avant les gratte-ciel, paru aux éditions France-Empire en 1979.

Notes et références

Liens externes

 Base Léonore
 Site de l'association Musiciens entre Guerre et Paix : biographies et photographies de Maréchal, Caplet, Durosoir
 Photographies de Coulibœuf, Cloëz, Durosoir, Magne, et Maréchal avec « le Poilu »
 Maurice Maréchal, portraits et documents sur Gallica

Violoncelliste français
Violoncelliste classique
Élève du conservatoire à rayonnement régional de Dijon
Officier de la Légion d'honneur
Personnalité liée à la musique classique décorée de la Légion d'honneur
Titulaire de la croix de guerre 1914-1918
Naissance à Dijon
Naissance en octobre 1892
Décès dans le 16e arrondissement de Paris
Décès en avril 1964
Décès à 71 ans

redpajama_set_name: RedPajamaWikipedia


Two critical aspects of proving (discriminatory) workplace retaliation for filing a workers compensation claim or engaging in other protected activity are: (1) proving causal relationship between the filing of the the workers compensation claim (or engaging in other protected activity) and the discriminatory actions by the employer; and (2) proving tat the employee was qualified and able to perform his or her job at the time when discrimination took place.
Careful investigation of the facts and circumstances of the employee's employment and wrongful termination is essential. Obviously, with respect to establishing the causal relationship, the closer in proximity of time the discriminatory conduct is to the employer's learning of the employee's filing a workers compensation claim, the more likely the causal link will seem to the judge, jury, or an arbitrator.
The first very important component of proving retaliation is in disproving a possible legitimate reason for termination, the most common of which is poor performance. Thus, in a case of alleged retaliation, like in most other discrimination claims, an attorney should seek evidence such as the employee's productivity and performance records, including all evaluations, commendations, promotions, raises, and any disciplinary measures.
The second component of demonstrating unlawful retaliation is the evidence specific to the workers' compensation system. Information to obtain of this type includes the number and disposition of all workers' compensation claims that have been filed against the employer, and information about the whereabouts and subsequent histories of employees who had filed claims.
The third piece of evidence essential to proving retaliation claim is the aggrieved employee's hospital and medical records, with special attention to the timing of visits to health care providers. The employer may have likely retaliated for an employee taking time off to see a doctor or get treatment, which in many cases is also considered an unlawful retaliation and disability discrimination.
It is also important to obtain documents evidencing the employer's position with respect to the underlying workers' compensation claim. i.e. Was your workers compensation claim contested by the employer? On what grounds? How vigorously? What was the outcome?
It is hard to overestimate the importance of documentary evidence in proving retaliation and disability discrimination claims in California. As in most individual discrimination claims, the testimony of the employee and employer generally ends up in flat contradiction regarding key elements of the claim. Proper documentation is likely to be crucial in corroborate the employee's testimony and disprove the testimony of the employer, casting doubt on the truthfulness of the employer's stated motives for discriminating and/or terminating your employment.


text:
Florida Whistleblower Lawyers > Blog > Healthcare Billing Fraud > The Stark Law And Whistleblower Claims
The Stark Law And Whistleblower Claims
Healthcare billing fraud is a significant problem in the United States. It occurs when providers in the medical industry—hospitals, physicians, clinics, pharmacies, etc—improperly bill private insurance companies or government programs. For health providers that accept government dollars (Medicare, Medicaid, etc), there are especially strict billing regulations in place.
The Stark Law refers to a set of federal regulations that prohibit self-referrals (kickbacks) in public health billing. Stark Law violations carry stiff penalties—and the government often relies on whistleblowers to uncover improper billing practices. Here, our Florida healthcare billing fraud lawyers highlight the key things you should know about the Stark Law and whistleblower claims.
Stark Law: Restrictions on Self-Referrals
A significant portion of all healthcare costs in the United States are covered by Medicare or Medicaid. The Stark Law bars doctors, hospitals, and other medical providers from billing Medicare or Medicaid based on a "self-referral." In effect, it is unlawful for a doctor (or other entity) to refer a patient to another provider with which they or their immediate family member has a direct financial interest.
Whistleblower Rights: The Stark Law and the False Claims Act
The federal government is responsible for enforcing the Stark Law. However, as with many other types of financial fraud, Stark Law violations are often difficult for authorities to identify. For this reason, the government often relies on whistleblower disclosures to help stop illegal self-referrals, medical industry kickbacks, and other types of healthcare billing fraud.
Through the False Claims Act, a whistleblower can take action to stop a Stark Law violation. A False Claims Act lawsuit can be filed directly by an individual whistleblower against a doctor, hospital, or any other party in violation of the Stark Law. A successful qui tam lawsuit allows a whistleblower to obtain an award constituting a portion of the compensation recovered by the government.
Stark Law and Whistleblower Example: A $115 Million Settlement
The cost of medical services is notoriously expensive. For this reason, many Stark Law violations can be significant. As an example, in 2015, the Department of Justice (DOJ) reached a $115 settlement in a Stark Law whistleblower case. The False Claims Act qui tam lawsuit was filed against Adventist Health System, over alleged Stark Law violations. The company settled the matter after facing accusations that it employed physicians who received direct financial bonuses for making certain referrals that resulted in a prohibited financial benefit for the company.
Notably, the enforcement action by the DOJ occurred after three whistleblowers came forward and filed two separate False Claims Act lawsuits against the company. The federal government, multiple state governments, and the whistleblowers all received an undisclosed share of the $115 million settlement.
Contact Our Florida False Claims Act Attorneys for Immediate Help
At Guttman, Freidin & Celler, our Florida healthcare billing fraud attorneys are strong, effective advocates for whistleblowers. If you have any questions about the Stark Law and whistleblower claims, we can help. Contact us now for a strictly confidential consultation. From our office in Miami, we provide whistleblower representation in Miami-Dade County, Florida, and nationwide.
justice.gov/opa/pr/adventist-health-system-agrees-pay-115-million-settle-false-claims-act-allegations
By Guttman, Freidin & Celler | Posted on December 27, 2021
« Southwest Florida Fire Chief Files Whistleblower Complaint, Alleges COVID-19 Fraud
An Overview Of COVID-19 Fraud And Whistleblower Claims »
More on Healthcare Billing Fraud
Florida Medical Device Company Will Pay $16 Million To Resolve False Claims Act Whistleblower Claim
COVID 19 Fraud
IRS Whistleblower
Florida Nursing Home Quality of Care Violations
We Have Successfully Recovered Over $5 Billion For Our Clients
$1.6 billion ABBOTT LABS U.S. ex rel. McCoyd v. Abbott Laboratories Western District of Virginia
$1.04 billion GlaxoSmithKline U.S. ex rel. Graydon v. GlaxoSmithKline District of Massachusetts
$2.3 billion Pfizer U.S. ex rel. Doghramji, et al. v. Community Health Systems Inc., et al. Eastern District of Pennsylvania
$97 million Community Health Systems U.S. ex rel. Doghramji, et al. v. Community Health Systems Inc., et al. Middle District of Tennessee
Address: 2 South Biscayne Boulevard, Suite 3100, Miami, FL 33131


text:
Late night socials during the Quiksilver Pro, Gold Coast
Your Stab correspondent has just returned from the Gold Coast of Australia, where the Quiksilver Pro, with pauses, took the the entirety of it's waiting period to complete. Due to the size of Australia's largest surf contest, late February every year sees the Coolangatta area injected with energy. Flags bearing the contest's logo hang from telegraph poles, large […]
style // Feb 22, 2016
Your Stab correspondent has just returned from the Gold Coast of Australia, where the Quiksilver Pro, with pauses, took the the entirety of it's waiting period to complete. Due to the size of Australia's largest surf contest, late February every year sees the Coolangatta area injected with energy. Flags bearing the contest's logo hang from telegraph poles, large billboards inform that the Quiksilver Pro is the "Best" of everything and every decent eatery in the area hosts who's-who lunch and dinner meets daily. The waves were, for a week straight, miniature. While swell on the points was non-existent during the lay-days, Duranbah, though barely pushing two foot on set waves, was overrun with male and female pros, as well as juniors, team managers, and free-surfers. And photogs. And filmers. It seemed as though the whole surf industry was in Coolangatta – even sponsored surfers who weren't in the contest were at every party and premiere, every night. At night-time during the Quiksilver Pro, the general consensus is to meet at Komune Beach Apartments (as nearly every night an event or party is held there), before moving on to something else. Here's the brief rundown of a coupla events that Stab attended:
– Josh Kerr's premiere for his new biopic, Kerrazy Kronicles, held at Pointbreak bar and restaurant, Burleigh Heads. The windows of the venue were mostly closed to fend off the powering wind, which turned the room into a heat-box. Dean Morrison wiped sweat from his brow, put his hand back on the crutches that support his injured leg, and used his other hand to lift a vodka, lime and soda to his mouth. A sickly-looking Mickey Avalon, dressed in patterned boardshots and a very badly-matched button-down shirt, drank wine and flicked greasy curls out of his face. Jordy Smith, taller than everyone else in the room, replied "Just water, bru", when his bud 'Laces' asked him what he wanted to drink. The film was typical Josh Kerr – modest, fun, with huge airs and incredible waves. Some of the waves were the more jealousy-inspiring seen in recent surf film vintage. The salt and pepper squid with Ponzu sauce was a hit. The room was made up of an 80/20 male/female ratio and many of the males mentioned the fact. Adam Bennetts used an Akai MPC sample pad and Ableton midi-controller to mix music. Kerr's infallible smile all evening gave no indicator of the furious manner in which he'd stomp through the media area of the contest a week later, having lost to Ace Buchan, and understandably mutter when asked for comment: "Nah, I'm pretty much fucking over it right now, ay."
– The Innersection premiere, held at Komune Beach Club at Greenmount. A runway stage sat across the water of the hotel's pool, and film conceptualists Taylor Steele and Nathan Myers stood on it with a microphone. Matt Meola and Craig Anderson were both asked onstage to stand in-between Taylor and Nathan, while the co-creators discussed the difficulty in choosing which of the two parts had earned the $100k prize-purse. Matt and Craig were visibly nervous, though as we've already said, Matt's presence was almost enough to confirm who the winner'd be before it was announced – kid didn't fly over from Maui for nothing. The tension in the moments leading up to the announcement was palpable. It was then, while his "Hero" section played in the background, that Matt Meola was awarded a $100k cheque to an eruption of applause. People with blue wristbands drank free Corona, everyone spoke about how Craig Anderson nearly won and whether they should, following the premiere, go to Dion Agius' premiere at Neverland or go to the Roxy party. Stab chose to go to Dion Agius' premiere. Surprised? Didn't think so.
– The premiere for Dion Agius' film, Thanks For The Dinner And The Sex, a low-budget, self-directed and self-edited piece by Dion, held at Neverland nightclub. The upstairs area of the club was packed with people. It was dark, sweaty, naughty and fun. Jordy Smith, again, didn't drink. This is worth noting, due to Jords' usual party-boi rep. Has the kid grown into a mature man, ready to sacrifice a few good times for a shot at the title? Time will tell. Kelly Slater and girlfriend Kalani turned up late, sat in a corner for a while, and inflicted the most intense presence imaginable on the entire room. Kelly sat with his back to the wall, staring with piercing blue eyes through the club, while every other person in the room did a bad job of pretending not to look at him. The movie: Indie gold (home-movie vibe), unselfish (first section – no Dion), surprising (for the haters – a full section of Dion "fly-boy" Agius doing only turns – and good ones), and aurally-pleasing (killer soundtrack, as per the man's taste.)
– Oh, then there was the party at Komune that we ourselves threw with our pals at Nike 6.0, titled A Warm-Hearted Hustle. A 10 metre-high image of Julian Wilson grabbing method and throwing a peace sign greeted attendees, and Laura Enever signature cocktails were served. In the corner sat a messy bed, acting as a kinky photo-booth where people 'posed'. As is the tendency at these things, the extensive bar tab had been demolished an hour before the party was due to finish. For photos of the night, jump over here.
– The Quiksilver Pro, New York launch party, held at Neverland Nightclub. The premise of this event was, if nothing else, confusing. The NY event ain't till September but Quik threw a launch-party in March. Hey, any excuse, right? On the way there I was told that a black wristband was required for upstairs entry, so I called Quik-rider Craig Anderson to ask if he had any spares. As he was leaving early, he had the driver of the car he was in pull-over to meet me and give me his torn-apart plastic bracelet. Admittedly I spent most of the night downstairs watching Dion Agius DJ – he was asked to play some music by the club owner, and obliged with the Rolling Stones, the Beatles and a bunch of other stuff that had the crowd more excited than the 'club music' played by the following DJ. It's interesting how strangely the crowd patterns in Neverland move. One minute, it's shoulder-to-shoulder and ordering a drink becomes a rib-crushing, voice-cracking exercise. The next moment, there's no more than 20 people in the downstairs bar, as everyone's decided to relocate upstairs. The migration cycle re-occurs every 15 minutes. Such is pack mentality.


text:
Features Khon Kaen International Marathon
01 April 2008, 7am
Distance Running 2008 April–June > Features > Marathon flowering in Thailand
The strong community involvement evident along the course is an attractive feature of the race
Marathon flowering in Thailand
by Jacky Knowles
Khon Kaen International Marathon
Sun 27 January 2008
Khon Kaen is a town of 150,000 people located 400km north-east of Bangkok, and has long been famous for silk weaving, dinosaur remains, sticky rice and spicy papaya salad, but it is now cultivating a new reputation as the "Marathon City" of Thailand.
The Khon Kaen International Marathon has been held at the end of January for the past five years and is increasing in size and status with each edition.
Race morning this year turned out to be around 4ºC warmer than for the previous year's event, yet it was still reasonably cool for this part of Asia. There was also a brief early morning sprinkle of rain for the 588 marathon runners as they prepared for their 04.30 start at the front of Khon Kaen University. The marathon route made a circuit of the city, with dawn arriving shortly after competitors passed the impressive nine-storey temple by the lake to the south of the city. Since the roads were all closed to traffic until the last runner had gone through, the run north through the city centre and along the ring road was a safe one.
After passing beside paddy fields full of bright green rice seedlings, the course headed slightly up and back towards the university. The last 5km was on a mainly downwards slope through the treelined roads of the university. Although there are no really prominent hills in the area, many runners noted after the race that few parts of the course are actually flat. Runners were buoyed along by students manning the frequent water stations, cheering, playing drums and waving flags to represent and welcome the twenty plus different nationalities of the runners taking part.
A full range of events from 4km fun run to the Marathon, each offering trophies and prize money for many different categories of finisher, attract participants from across the world. The strong community involvement evident along the course is another attractive feature of the race. Thousands of local residents and school children from neighbouring provinces ran the mini marathon (11.55 km), many of them doing so barefoot.
The half marathon start time and course were designed so that runners heading out to the half way point could be inspired by the sight of the lead group of mainly Kenyan and Ethiopian marathon runners heading for home in the opposite direction in a parallel lane. This same section of road boasted a stage with local singers, especially for the benefit of the runners. Local families also got involved by setting up tables with slices of watermelon at different points along the route.
The mini marathon runners made a circuit around and through the university, finishing in the same place as the other two longer events and thus benefiting from the same access to food and the much appreciated massage services. Finishers of all the three main events received the now-traditional weighty medal to commemorate and celebrate their efforts.
The Marathon was well contested at the front, with Kenya's Luka Kipkemoi Chelimo finishing less than 50 metres ahead of his compatriot, Geoffrey Kidrotich Birgen. Although many Kenyan entrants had not been able to make the journey due to the troubles in their country at that time, they were still well-represented and took nine out of the first 10 places. The first Thai runner to cross the line was Kordair Senmood in 13th place with a time of 2:35:15.
Thai women made an exceptionally strong showing, sweeping up eight of the top 10 prizes, including first place. Two hours into the race Norah Cheptoo from Kenya was leading the group but, following a strained hamstring, she dropped back to achieve an eventual fourth place. Sunisa Saylomyen, who had never been far behind, came through to win ahead of last year's winner Wioletta Uryga from Poland. Sunisa was building on her win at the South East Asia (SEA) Games just six weeks previously (and she went on to run 2:48 in Hong Kong on 17 Feb).
The Kenyan and Ethiopian runners dominated again in the men's half marathon, taking the first six places. An exciting last few hundred metres resulted in only three seconds separating Robert Kosgei of Kenya and Mekonnen Emiru from Ethiopia. The previous Thai marathon champion, Boonchu Chantorndesa crossed the line with his fellow countryman Amnuay Tongmitr in the same time (1:11:06) but Amnuay was awarded seventh place and Boonchu eighth, based on a 0.01 second difference in their times.
Tenth place went to a new competitor, Hem Bunting, from Cambodia. Mostly self-motivated and self-trained, he now lives in the office of the Khmer Amateur Athletics Federation (KAAF) along with his running mate, Cheng Chandara, who also ran in Khon Kaen and placed in the top 50 in the half marathon. Hem Bunting had recently competed in the SEA Games marathon, bringing home a silver medal for his national record breaking time of 2:26. Khon Kaen International Marathon provides a great opportunity for Cambodian and Lao athletes, for whom one of the biggest challenges is a lack of domestic, or otherwise affordable, race opportunities.
In the women's half marathon event, the first three runners reached the half way point shoulder to shoulder before Saifon Bunchaeng of Thailand pulled away to win.
The ever-popular 4km fun run was held on a separate route, going 2km up and back on the airport road from the south gate of the university. As in 2007, over 30,000 people took part, making for an impressive sight, especially since most of the participants wore the official yellow race T-shirt which represents the provincial flower of Khon Kaen. Some people raced the whole way, others strolled the course along with their pets, while most did something in between, in an effort to win one of the 20,000 finishers' medals.
By mid morning the post-race entertainment, consisting of different local and Chinese dances and the now customary cobra show, had finished and the awards and cash prizes had been presented. Organisers were left to enjoy a well-deserved rest while contemplating how to build on this successful event for next year (25 January 2009).
See the latest news from Khon Kaen International Marathon