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Susceptibility
Echinocandin Susceptibility Testing of Candida Species: Comparison of EUCAST EDef 7.1, CLSI M27-A3, Etest, Disk Diffusion, and Agar Dilution Methods with RPMI and IsoSensitest Media
Maiken Cavling Arendrup, Guillermo Garcia-Effron, Cornelia Lass-Flörl, Alicia Gomez Lopez, Juan-Luis Rodriguez-Tudela, Manuel Cuenca-Estrella, David S. Perlin
Maiken Cavling Arendrup
Unit of Mycology and Parasitology, Statens Serum Institut, Copenhagen, Denmark
For correspondence: mad@ssi.dk
Guillermo Garcia-Effron
Public Health Research Institute, UMDNJ-New Jersey Medical School, Newark, New Jersey
Cornelia Lass-Flörl
Department of Hygiene and Medical Microbiology, Innsbruck Medical University, Innsbruck, Austria
Alicia Gomez Lopez
Servicio de Micología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Spain
Juan-Luis Rodriguez-Tudela
Manuel Cuenca-Estrella
David S. Perlin
DOI: 10.1128/AAC.01256-09
This study compared nine susceptibility testing methods and 12 endpoints for anidulafungin, caspofungin, and micafungin with the same collection of blinded FKS hot spot mutant (n = 29) and wild-type isolates (n = 94). The susceptibility tests included EUCAST Edef 7.1, agar dilution, Etest, and disk diffusion with RPMI-1640 plus 2% glucose (2G) and IsoSensitest-2G media and CLSI M27A-3. Microdilution plates were read after 24 and 48 h. The following test parameters were evaluated: fks hot spot mutants overlapping the wild-type distribution, distance between the two populations, number of very major errors (VMEs; fks mutants misclassified as susceptible), and major errors (MEs; wild-type isolates classified as resistant) using a wild-type-upper-limit value (WT-UL) (two twofold-dilutions higher than the MIC50) as the susceptibility breakpoint. The methods with the lowest number of errors (given as VMEs/MEs) across the three echinocandins were CLSI (12%/1%), agar dilution with RPMI-2G medium (14%/0%), and Etest with RPMI-2G medium (8%/3%). The fewest errors overall were observed for anidulafungin (4%/1% for EUCAST, 4%/3% for CLSI, and 3%/9% for Etest with RPMI-2G). For micafungin, VME rates of 10 to 71% were observed. For caspofungin, agar dilution with either medium was superior (VMEs/MEs of 0%/1%), while CLSI, EUCAST with IsoSensitest-2G medium, and Etest were less optimal (VMEs of 7%, 10%, and 10%, respectively). Applying the CLSI breakpoint (S ≤ 2 μg/ml) for CLSI results, 89.2% fks hot spot mutants were classified as anidulafungin susceptible, 60.7% as caspofungin susceptible, and 92.9% as micafungin susceptible. In conclusion, no test was perfect, but anidulafungin susceptibility testing using the WT-UL to define susceptibility reliably identified fks hot spot mutants.
Three echinocandin class drugs, anidulafungin, caspofungin, and micafungin, are licensed for the treatment of invasive candidiasis. They are among the preferred agents for invasive candidiasis, as a number of recent fungemia surveys have reported a considerable proportion of cases involving species with reduced susceptibility to fluconazole (3, 4, 24, 28, 31, 37, 44). Additionally, anidulafungin has been associated with an improved success rate, even in cases involving fluconazole-susceptible species (39). Following increased use, sporadic cases of failures associated with elevated MICs have been reported. In the majority of cases, these failures have been associated with mutations in two hot spot regions of FKS genes, which encode the target and major subunit of the 1,3-ß-d-glucan synthase complex (5, 7, 22, 25, 26, 33, 34). Consequently, close monitoring and robust susceptibility testing methods have become increasingly important.
EUCAST and CLSI have developed standard methods based on broth dilution for the susceptibility testing of yeasts (9, 41). Methodological differences include glucose concentration, inoculum size, shape of microtiter wells (flat or round), and end-point reading (visual or spectrophotometric), but the methods are more alike than different and in general generate similar results (11, 42). Recently, CLSI proposed an S value of ≤2 μg/ml as a tentative susceptibility breakpoint for caspofungin, micafungin, and anidulafungin for Candida spp., taking into account analysis of mechanisms of resistance, an epidemiological MIC population distribution, parameters associated with success in pharmacodynamic models, and results of clinical efficacy studies (9, 38). As no significant differences in clinical response were noted among the various species, results for all species were merged, and a susceptibility breakpoint of 2 μg/ml was found to encompass the vast majority of isolates, while not bisecting the population of Candida parapsilosis. The crucial issue is whether current susceptibility testing methods and breakpoints clearly and reliably identify isolates with resistance mechanisms associated with treatment failures (5, 7, 8, 13, 14, 16, 18, 22, 25, 26, 33, 40). Not only have cases involving isolates classified as susceptible using the reference methods been shown to contain resistance mutations (5, 7, 13, 14, 22, 25), but also recent studies suggest that a breakpoint of an S value of ≤2 μg/ml may be too high for anidulafungin and micafungin, considering the 1,3-ß-d-glucan synthase kinetic inhibition data of wild-type and mutant enzymes from resistant strains (17, 18). Finally, we recently reported a resistant Candida albicans isolate that failed to be identified as resistant when the reference methodologies were used, while Etest, agar dilution, and disk diffusion methods correctly identified it (5).
We therefore undertook a comparative study of the two references methods, a modified EUCAST microdilution method using IsoSensitest medium, agar dilution, and disk and Etest diffusion using RPMI-1640 as well as IsoSensitest medium to evaluate their ability to reliably discriminate between a well-characterized panel of wild-type and fks hot spot mutant Candida isolates. The semisynthetic IsoSensitest medium was chosen as an alternative medium due to this medium having previously been shown to be appropriate for amphotericin B MIC testing (10).
Isolates.Totals of 94 clinical isolates and two reference strains (C. parapsilosis ATCC 22019 and C. krusei ATCC 6258) were used in this study, including 10 FKS wild-type and 10 fks hot spot mutant C. albicans isolates, 9 FKS wild-type and 11 fks hot spot mutant C. glabrata isolates, 1 FKS wild-type and 1 fks hot spot mutant C. dubliniensis isolate, 13 FKS hot spot wild-type and 3 fks hot spot mutant C. krusei isolates, 19 FKS wild-type C. parapsilosis isolates, and 15 FKS hot spot wild-type and 4 fks hot spot mutant C. tropicalis isolates. Three isolates were found to harbor mutations outside the resistance hot spots and were regarded as wild type concerning echinocandin susceptibility because of their normal kinetic inhibition properties. Thus, a total of 29 isolates with characteristic echinocandin resistance mutations in the hot spot regions were included (Table 1). Species identification was based on colony color and morphology on CHROMagar (CHROMagar Co., Paris, France), microscopic morphology on cornmeal agar (SSI Diagnostica, Hilleroed, Denmark), ability to grow at 37°C and 42°C, and use of a commercial system (ATB ID32C; bioMérieux, Marcy l'Etoile, France). All isolates were coded, and tests performed blinded for susceptibility patterns. Each susceptibility test method was performed in one of the four participating laboratories.
Amino acid substitutions within the Fks proteins from the Candida isolates studieda
Compounds.A single lot of pure substance for each of the three echinocandins was used throughout the study. Stock solutions were prepared in dimethyl sulfoxide (DMSO; Sigma) for anidulafungin (provided by Pfizer), in water for caspofungin (provided by Merck), and in water or saline for micafungin (provided by Astellas), taking into account the potencies of the powders. MICs obtained by the CLSI M27-A3 method fell within the recommended ranges for both control strains, thus confirming the activity of the powders.
Agar dilution.Twelve-well multidish plates (1.9 cm2/well) containing a twofold-dilution series of each of the three echinocandins (concentration range of 0.015 to 4 μg/ml, 1-ml volumes) and a drug-free well were prepared using RPMI 1640 2% glucose buffered with MOPS [morpholinepropanesulfonic acid] (RPMI-2G) and IsoSensitest 2% glucose (IsoSensitest-2G), respectively (Balis Laboratorium, Boven Leeuwen, The Netherlands). A 1 to 5 × 106-CFU/ml suspension of each Candida isolate was prepared and diluted 1:6,000, and 20 μl (containing 3 to 16 CFU) were plated on each agar surface. Plates were incubated at 37°C for 48 h. Growth was compared to that in the drug-free control well; wells with tiny microcolonies were ignored.
EUCAST microdilution.EUCAST microdilution was performed strictly according to the standard (EDef 7.1 [41]) and additionally using IsoSensitest-2G medium (Oxoid S.A., Madrid Spain). Plates were prepared in one batch, sealed in aluminum foil, and stored at −80°C for no longer than 2 months before use. Microtiter plates were read spectrophotometrically at 530 nm after 24 h as well as after 48 h, and the MIC was determined using 50 as well as 90% growth inhibition. C. krusei ATCC 6258 and C. parapsilosis ATCC 22019 were used as quality control strains through all experiments.
CLSI microdilution.CLSI microdilution was performed in triplicate strictly according to the CLSI M27-A3 standard (9) using three separate batches of plates prepared using the single lots of antifungal compounds. Plates were stored at −86°C for a maximum of 15 days before use. Microtiter plates were read visually, and the MIC was determined using prominent inhibition (corresponding to 50%) as an end point. An additional reading was performed after 48 h. Geometric mean values were determined, and the values between the log2 scale were adjusted to the upper closest log2 value for comparison. C. krusei ATCC 6258 and C. parapsilosis ATCC 22019 were used as quality control strains through all experiments.
Etest and disk diffusion testing.Susceptibilities of the various isolates to anidulafungin, micafungin, and caspofungin were determined by Etest (AB Biodisk, Solna, Sweden) using RPMI-2G agar (Sigma, Vienna, Austria) according to the manufacturer's recommendations and using IsoSensitest-2G (Difco, Vienna, Austria). For disk diffusion testing, 90-mm plates containing RPMI-2G agar and IsoSensitest-2G (Difco, Vienna, Austria), respectively, were inoculated by swabbing the agar with a swab soaked in a yeast suspension of 1 × 106 to 2.5 × 106 cells/ml. Disks containing 5 μg of drug were prepared by placing 20 μl of the appropriate drug concentration on sterile 6-mm-diameter paper disks and subsequently placing the disks on the inoculated plates. Stock solutions were prepared by dimethyl sulfoxide and sterile water (Sigma, Vienna, Austria) as described above. Etests and disk diffusion were read at 24 h. The Etest MICs were rounded up to the next even log2 concentration for comparison. For disk diffusion, the zone diameters were evaluated in mm.
FKS gene sequence analysis.Candida sp. genomic DNA was extracted from yeast cells grown overnight in YPD (yeast extract, 2%; BactoPeptone, 4%; dextrose, 4%) broth medium with a Q-Biogene (Irvine, CA) FastDNA kit. PCR and sequencing primers were designed based on the C. albicans FKS1 gene (GenBank accession no. XM_716336), C. glabrata FKS1 and FKS2 (GenBank accession no. XM_446406 and XM_448401, respectively), C. tropicalis FKS1 (GenBank accession no. EU676168), C. krusei FKS1 (GenBank accession no. EF426563), C. dubliniensis FKS1 (GenBank accession no. GQ342611), and C. parapsilosis FKS1 (GenBank accession no. EU221325). The hot spot regions of the FKS genes (35) of all the strains included in this study were sequenced with a CEQ dye terminator cycle sequencing quick start kit (Beckman Coulter, Fullerton, CA) according to the manufacturer's recommendations. Sequence analysis was performed with CEQ 8000 genetic analysis system software (Beckman Coulter, Fullerton, CA) and BioEdit sequence alignment editor (Ibis Therapeutics, Carlsbad, CA). The specific mutations are shown in Table 1.
Evaluation of test performance.For each of the drug-bug combinations, the following parameters were used to evaluate and compare the test performances: distance between end-point ranges (MIC or zone diameter) for fks hot spot mutant isolates and wild-type isolates calculated as number of twofold-dilution steps (for MIC results) and number of millimeters separating the zones (for disk diffusion) (negative values indicate the degree of overlap expressed as the number of dilution steps or millimeters involved in the overlap); overlap, which is defined as the number of end points for the fks hot spot mutant isolates that overlapped with the end-point range of the wild-type populations; wild-type-upper-limit values (WT-UL), which for MIC tests is defined as two twofold-dilution steps higher than the MIC50 (if the population was truncated with all MICs at or below the lowest concentration tested, the WT-UL was defined as two times the lowest dilution tested); very major errors (VME), which is the number of fks hot spot mutant isolates with a MIC lower or equal to the WT-UL; and major errors (ME), which is the number of wild-type isolates with a MIC above the WT-UL.
For each susceptibility test, 94 isolates were tested against the three candins (282 test results per test). Twenty-nine isolates had FKS mutations in the hot spot regions, leading to 87 test results for such isolates per test.
Reference methods.Overall wild-type and fks hot spot mutant populations were separated by −1 (the negative value representing an overlap) to 6 twofold-dilution steps using EUCAST (Table 2). The MIC ranges of wild-type and fks hot spot mutant isolates overlapped for 2 of 15 drug-Candida sp. combinations, for anidulafungin and C. krusei and for micafungin and C. glabrata, involving 2/84 test results for fks hot spot mutant isolates (one isolate failed to grow) (Tables 2 and 3). CLSI overlaps between wild-type and fks hot spot mutant populations were seen for four drug-Candida sp. combinations, anidulafungin and C. tropicalis, caspofungin and C. tropicalis, micafungin and C. glabrata, and micafungin and C. tropicalis, involving 5/84 test results for fks hot spot mutant isolates (Tables 3 and 4). Overall, wild-type and fks hot spot mutant isolates were separated by −2 to 5 twofold-dilution steps. Applying the CLSI breakpoint for susceptibility of a MIC of ≤2 μg/ml, 89.2% (25/28) isolates with FKS hot spot mutations were classified as anidulafungin susceptible, 60.7% (17/28) as caspofungin susceptible, and 92.9% (26/28) as micafungin susceptible by the CLSI reference method.
MICs obtained using the EUCAST EDef 7.1 (RPMI-2G and 50% inhibition)a
Comparison of the following test performance parameters: the proportion of MIC ranges for the 29 fks hot spot mutant isolates overlapping the MIC range for wild-type isolates, the proportion of VMEs in the test results for fks hot spot mutant isolates; and the proportion of MEs in the test results for wild-type isolates when applying the WT-ULs as species-specific susceptibility breakpointsa
MICs obtained using CLSI M27-A3 (RPMI-2G and 50% inhibition)a
Modified reference methods.Microdilution testing was also performed according to EUCAST Edef 7.1 but using IsoSensitest-2G as a test medium (Table 5). Overall wild-type and fks hot spot mutant populations were comparably separated by −1 to 5 dilution steps. The MIC ranges of wild-type and fks hot spot mutant isolates overlapped for two drug-Candida sp. combinations, caspofungin and C. krusei and micafungin and C. glabrata, thus involving 4/87 test results for fks hot spot mutant isolates (Table 3 and 5). Furthermore, EUCAST and CLSI microdilution trays were evaluated after 48 h. However, this led to poorer discrimination between wild-type and fks hot spot mutant isolates (Table 3).
MICs obtained using a modified EUCAST methodology (IsoSensitest-2 and 50% inhibition)
Agar diffusion methods.Susceptibility testing was performed using Etest and 5-μg disks with RPMI-2G medium, as well as IsoSensitest-2G medium as a test agar. By Etest using RPMI-2G (as recommended by the manufacturer), the distances between wild-type and fks hot spot mutant isolates were −4 to 8 dilution steps, and overlapping populations were seen for three drug-Candida sp. combinations, anidulafungin and C. krusei, caspofungin and C. krusei, and micafungin and C. glabrata, involving 4/87 test results for fks hot spot mutant isolates (Tables 3 and 6). When the CLSI breakpoint was applied, 82.8% (24/29) of the isolates with FKS mutations were classified as anidulafungin susceptible, 44.8% (13/29) as caspofungin susceptible, and 96.6% (28/29) as micafungin susceptible. Replacing the medium with IsoSensitest-2G did not improve the performance; thus, overlapping populations were seen for seven drug-Candida sp. combinations involving 14/87 test results for fks hot spot mutant isolates (Tables 3 and 7). By disk test using RPMI-2G agar, overlaps between wild-type and fks hot spot mutant isolates were seen for 9/15 drug-Candida sp. combinations, involving 18/87 test results for fks hot spot mutant isolates (Tables 3 and 8). Using disks and IsoSensitest-2G agar, overlaps between wild-type and fks hot spot mutant isolates were seen for 9/15 drug-Candida sp. combinations, involving 14/87 test results for fks hot spot mutant isolates and 1/195 results for wild-type isolates (Tables 3 and 9).
MICs obtained using Etest and RPMI-2G agar
MICs obtained using Etest and IsoSensitest-2 agar
Zone diameters obtained using 5-μg disks and RPMI-2G agara
Zone diameters obtained using 5 μg disks and IsoSensitest-2% glucose agara
Agar dilution.Agar dilution performed using RPMI-2G resulted in a distance between the wild-type and fks hot spot mutant populations of −1 to ≥5 (Table 10), and overlapping populations were seen for three drug-Candida sp. combinations, anidulafungin and C. tropicalis, caspofungin and C. tropicalis, and micafungin and C. tropicalis, involving 2/87 test results for fks hot spot mutant isolates and 1/195 test results for wild-type isolates (Tables 3 and 10). Performing agar dilution with IsoSensitest-2G did not improve discrimination between wild-type and fks hot spot mutant isolates. Thus, the distances between wild-type and fks hot spot mutant isolates was −1 to >3 dilution steps, and overlaps between wild-type and fks hot spot mutant isolates were seen for five drug-Candida spp. combinations, involving 7/87 test results for fks hot spot mutant isolates (Tables 3 and 11).
TABLE 10.
MICs obtained by agar dilution using RPMI-2G agara
MICs obtained by agar dilution using IsoSensitest-2 agar
WT-UL.Defining susceptibility as a MIC ≤ the WT-UL value, the proportions of fsk hot spot mutants being misclassified as susceptible (VMEs) and MEs (susceptible isolates being classified as resistant) were calculated and presented together with the WT-ULs for each test, compound, and species in Tables 2, 4 to 7, 10, and 11. An overview of VMEs and MEs per test and Candida species is presented in Table 3. The lowest numbers of VMEs and MEs across the three echinocandins were seen for susceptibility testing using CLSI M27A-3 (12% and 0%, respectively), agar dilution with RPMI-2G medium (14% and 0%), and Etest with RPMI-2G medium (8% and 3%). The fewest VMEs and MEs were observed overall for anidulafungin with EUCAST (4% and 1%), CLSI (4% and 3%), and Etest with RPMI-2G (3% and 9%). For micafungin, VME rates of 10 to 71% were observed, and notably both reference tests resulted in a quarter to a third of the fks mutants being misclassified as susceptible (Table 3). For caspofungin, agar dilution with either medium was notably superior, leading to no VMEs and a 1% ME rate, while CLSI, EUCAST with IsoSensitest-2G medium, and Etest were less optimal, leading to VME rates of 7%, 10%, and 10%, respectively, and no MEs.
C. parapsilosis.The wild-type isolates of C. parapsilosis separated clearly from the other species for anidulafungin and micafungin using microdilution tests and agar dilution, with an epidemiological cut-off value of 2 to 8 μg/ml (Tables 2, 4, 5, 10, and 11). By Etest and disk diffusion, the ranges of MICs/zone diameters were very wide, suggesting low reproducibility and variability of the results (Tables 6 to 9).
This study compared the major susceptibility testing methods by evaluating a significant number of fks hot spot mutant and wild-type isolates with nine different tests and 12 different end points on the same collection of blinded isolates. Direct comparison of MICs obtained by the different methods is problematic, as the tests were performed in different laboratories. However, for each test it is possible to evaluate the potential for discriminating fks hot spot mutants from wild-type isolates and then to compare the performances of the individual tests with respect to this crucial parameter, expressed as distance between the populations and number of overlaps, VMEs, and MEs.
When the tests were ranked according to the number of fks hot spot mutant isolates overlapping the susceptible population, agar dilutions using RPMI-2G and EUCAST EDef 7.1 were found to be best at discriminating wild-type isolates from isolates with mutations in the FKS hot spots. EUCAST with IsoSensitest-2G medium, CLSI M27-A3, and Etest using RPMI-2G performed slightly less well, and the remaining tests suffered from a high number of overlaps between isolates with resistance mutations and wild-type isolates. For the agar dilution tests, each wild-type distribution typically spanned 3 log2 dilution steps, in agreement with what we have previously published using one batch of trays in a single lab (6). When results from a large number of isolates, several technicians and batches of media, many laboratories, and other necessarily allowed variations in testing conditions are taken into account, the base of the perceived wild-type distribution broadens to a maximum of five dilution steps, with the MIC50 in the middle and the epidemiological cut-off value (ECOFF) two twofold-dilution steps higher (6). We similarly established species- and drug-specific WT-ULs, applying this principle to the limited number of wild-type isolates included in this study, as a comparison of the numbers of fks hot spot mutants with MICs lower than or equal to the WT-UL value is likely a better predictor of the test's ability to pick up mutant isolates when performed in different laboratories, using different batches of trays, etc.
The lowest numbers of VMEs and MEs across the three echinocandins seen seen for susceptibility testing using CLSI M27A-3, agar dilution, or Etest with RPMI-2G medium, but still ∼10% of the fks hot spot mutants were classified as susceptible. The consequence of this is that even with the best performing assays and a susceptibility breakpoint set as low as the WT-UL, 10% of the fks mutants are misclassified as susceptible. If, on the other hand, the susceptibility breakpoint is defined even lower in order to ensure that fks mutants are classified as nonsusceptible, a number of susceptible isolates will be classified as potentially resistant. The first scenario may not currently affect a great number of cases, as fks mutants are still only sporadically reported, but for individual patients, the consequence may be inappropriate treatment and failure (5, 7, 8, 22, 25, 26, 33, 45). The other scenario will affect a greater number of cases, for which further analyses are needed in order to determine if the isolate is truly resistant or not.
Considerable differences in test performances were seen for the different echinocandins. Caspofungin agar dilution with either medium was superior, in agreement with previous observations (5). For micafungin, in general all micro- or agar dilution tests led to a misclassification of at least a quarter of the fks hot spot mutant isolates. This was due mainly to C. glabrata isolates with resistance mutants not separating very well from the wild-type population (17, 45). Etest with RPMI-2G performed best for micafungin, but still a quarter of the fks mutant C. glabrata isolates were misclassified as susceptible. This raises the question of whether a number of resistant isolates may go unnoticed when current reference methodologies for susceptibility testing are used.
Anidulafungin was the compound for which the fewest VMEs were observed overall. This may suggest that susceptibility testing for this agent could be used as a screening procedure for echinocandin susceptibility. In this study, anidulafungin susceptibility testing by any of the current reference methodologies or Etest with RPMI-2G resulted in misclassifications of 3 to 4% of fks hot spot mutant isolates when the species-specific WT-ULs indicated in Tables 2, 4, and 6 were used as susceptibility breakpoints. These WT-UL values are considerably lower than the currently recommended CLSI breakpoint for susceptibility of an S value of ≤2 μg/ml (9, 38). However, the vast majority of isolates with FKS hot spot mutations were classified as susceptible by CLSI and Etest using the CLSI breakpoint, in agreement with the increasing concern that the breakpoint may be too high (5, 7, 13, 14, 18, 22, 25). This was particularly true for anidulafungin and micafungin, in agreement with the fact that the MICs of these compounds are in general lower than that of caspofungin and much lower than the breakpoint (12, 29, 30). Further studies are needed to establish ECOFFs and breakpoints, reducing the risk of misclassification of resistant isolates.
The MICs of all three candins were considerably higher for C. parapsilosis than for the other species, as previously described (12, 29, 36, 38). C. parapsilosis has a naturally occurring mutation at hot spot 1 in FKS1, accounting for the MIC level being in the same range as those of hot spot mutant isolates of the other Candida species (16). The overall clinical response for invasive C. parapsilosis infections treated with echinocandins is comparable to that for the other species (23, 27, 32, 38), which may in part be due to C. parapsilosis being less virulent (1, 2, 19-21, 46). However, C. parapsilosis was the only species for which the response rate observed with patients with invasive candidiasis receiving anidulafungin was numerically but not statistically significantly poorer than that for fluconazole (response rates of 64% versus 88%) (39), and breakthrough infections with C. parapsilosis in hematological patients have been associated significantly with prior caspofungin use, suggesting that other antifungal drug classes may be more efficacious, although this fungus is not clinically highly resistant (15, 43). These observations suggest that species-specific breakpoints and lower breakpoints than the current CLSI ones are necessary to avoid classifying C. albicans, C. dubliniensis, C. glabrata, and C. tropicalis isolates with resistance mutations as susceptible while not classifying all wild-type C. parapsilosis isolates as resistant.
When the MICs obtained for the three candins were compared, it was noted that the MICs obtained by EUCAST EDef 7.1 were in general higher for caspofungin than those obtained using the CLSI M27-A3 method. And for both tests the MIC50s were higher than those normally obtained (one or two dilutions for CLSI and two dilution steps for EUCAST) (36). The reason for this is not clear, but it indicates a variability associated with caspofungin susceptibility testing, despite performing testing in reference laboratories using a single batch of pure substances and correcting for potency, as we have summarized previously (5). The implication of this for the current study is that WT-UL values and MIC ranges might be lower if repeated with a different batch of pure substance, but it is not likely that it affects the evaluation of the performances of the tests and their ability to differentiate fks hot spot mutants from wild-type isolates. Further studies of caspofungin testing are needed to resolve this issue.
In conclusion, no test was perfect, in the sense that MIC ranges for fks mutants and wild-type isolates were separated by at least three twofold-dilution steps and no overlap existed between wild-type and fks hot spot mutant populations for all three candins and the Candida species included. Until the challenges concerning caspofungin testing reproducibility and micafungin susceptibility testing of C. glabrata are solved, an attractive approach might be to use anidulafungin MICs greater than the species-specific ECOFFs as a marker for possible echinocandin resistance. For this purpose, EUCAST with RPMI-2G or IsoSensitest-2G, CLSI, and Etest are all appropriate methods, while disk testing performed poorly. Possibly resistant isolates should be confirmed and the underlying mechanism investigated by sequence analysis of the FKS hot spot regions. Lack of mutations may represent a falsely elevated MIC during initial testing or resistance due to yet uncharacterized mechanisms.
We thank Birgit Brandt for excellent technical assistance. We thank Francoise Dromer, Unité de Mycologie Moléculaire, Institut Pasteur, Paris, France, for providing one of the candin fks hot spot mutant C. krusei isolates (13). We thank Astellas for providing micafungin pure substance, Merck for providing caspofungin pure substance, and Pfizer for providing anidulafungin pure substance.
The study was financially supported in part by research grants from the Investigator-Initiated Study programs of Astellas Pharma, Merck & Co. Inc., and Pfizer Aps. Additionally, it was supported by NIH grant AI069397 to D.S.P.
The opinions expressed in this paper are those of the authors and do not necessarily represent those of the pharmaceutical companies.
Potential conflicts of interest are that M.C.A. has been a consultant for Astellas, Merck, Pfizer, and SpePharm and an invited speaker for Astellas, Cephalon, Merck Sharp & Dohme, Pfizer, Schering-Plough, and Swedish Orphan and has received research funding for this particular study from Astellas, Merck, and Pfizer.
C.L.-F. has been a consultant for Pfizer and Schering Plough and an invited speaker for Pfizer, Gilead, Schering Plough, and Merck Sharp & Dohme and has received research funding from Pfizer, Gilead, Merch Sharp & Dohme, and Schering Plough.
D.S.P. is a shareholder in Merck; has acted as a consultant for Merck, Pfizer, and Astellas; is an advisory board member for Merck, Pfizer, Astellas, and Myconostica; has a patent for assays for resistance to echinocandin-class drugs (application no. 07763-O69WO1); has received research funding (although not for this particular study) from Merck, Pfizer, Astellas, and Myconostica; and has been an invited speaker for Merck, Pfizer, Astellas, and Myconostica.
G.G.-E. has no conflicts of interest.
A.G.L. has no conflicts of interest.
In the past 5 years, J.L.R.T. has received grant support from Astellas Pharma, Gilead Sciences, Merck Sharp & Dohme, Pfizer, Schering Plough, Soria Melguizo SA, the European Union, the Spanish Agency for International Cooperation, the Spanish Ministry of Culture and Education, the Spanish Health Research Fund, the Instituto de Salud Carlos III, the Ramon Areces Foundation, and the Mutua Madrileña Foundation. He has been an advisor/consultant to the Panamerican Health Organization, Gilead Sciences, Merck Sharp & Dohme, Myconostica, Pfizer, and Schering Plough. He has been paid for talks on behalf of Gilead Sciences, Merck Sharp & Dohme, Pfizer, and Schering Plough.
In the past 5 years, M.C.E. has received grant support from Astellas Pharma, BioMerieux, Gilead Sciences, Merck Sharp & Dohme, Pfizer, Schering Plough, Soria Melguizo SA, the European Union, the ALBAN program, the Spanish Agency for International Cooperation, the Spanish Ministry of Culture and Education, the Spanish Health Research Fund, the Instituto de Salud Carlos III, the Ramon Areces Foundation, and the Mutua Madrileña Foundation. He has been an advisor/consultant to the Panamerican Health Organization, Gilead Sciences, Merck Sharp & Dohme, Pfizer, and Schering Plough. He has been paid for talks on behalf of Gilead Sciences, Merck Sharp & Dohme, Pfizer, and Schering Plough.
Received 4 September 2009.
Returned for modification 10 October 2009.
↵▿ Published ahead of print on 2 November 2009.
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Arendrup, M. C., K. Fuursted, B. Gahrn-Hansen, I. M. Jensen, J. D. Knudsen, B. Lundgren, H. C. Schonheyder, and M. Tvede. 2005. Seminational surveillance of fungemia in Denmark: notably high rates of fungemia and numbers of isolates with reduced azole susceptibility. J. Clin. Microbiol. 43:4434-4440.
Arendrup, M. C., K. Fuursted, B. Gahrn-Hansen, H. C. Schonheyder, J. D. Knudsen, I. M. Jensen, B. Bruun, J. J. Christensen, and H. K. Johansen. 2008. Semi-national surveillance of fungaemia in Denmark 2004-2006: increasing incidence of fungaemia and numbers of isolates with reduced azole susceptibility. Clin. Microbiol. Infect. 14:487-494.
Arendrup, M. C., G. Garcia-Effron, W. Buzina, K. L. Mortensen, N. Reiter, C. Lundin, H. E. Jensen, C. Lass-Florl, D. S. Perlin, and B. Bruun. 2009. Breakthrough Aspergillus fumigatus and Candida albicans double infection during caspofungin treatment: laboratory characteristics and implication for susceptibility testing. Antimicrob. Agents Chemother. 53:1185-1193. doi:0.1128/AAC.01292-08.
Arendrup, M. C., G. Kahlmeter, J. L. Rodriguez-Tudela, and J. P. Donnelly. 2009. Breakpoints for susceptibility testing should not divide wild-type distributions of important target species. Antimicrob. Agents Chemother. 53:1628-1629. doi:10.1128/AAC.01624-08.
Baixench, M. T., N. Aoun, M. Desnos-Ollivier, D. Garcia-Hermoso, S. Bretagne, S. Ramires, C. Piketty, and E. Dannaoui. 2007. Acquired resistance to echinocandins in Candida albicans: case report and review. J. Antimicrob. Chemother. 59:1076-1083.
Cleary, J. D., G. Garcia-Effron, S. W. Chapman, and D. S. Perlin. 2008. Reduced Candida glabrata susceptibility secondary to an FKS1 mutation developed during candidemia treatment. Antimicrob. Agents Chemother. 52:2263-2265.
Clinical and Laboratory Standards Institute (CLSI). 2008. Reference method for broth dilution antifungal susceptibility testing of yeasts, 3rd ed. Approved standard. CLSI M27-A3(28). Clinical and Laboratory Standards Institute, Wayne, PA.
Cuenca-Estrella, M., T. M. Diaz-Guerra, E. Mellado, and J. L. Rodriguez-Tudela. 2001. Detection of resistance to amphotericin B in Candida isolates by using Iso-Sensitest broth. Antimicrob. Agents Chemother. 45:2070-2074. doi:10.1128/AAC.45.7.2070-2074.2001.
Cuenca-Estrella, M., W. Lee-Yang, M. A. Ciblak, B. A. Arthington-Skaggs, E. Mellado, D. W. Warnock, and J. L. Rodriguez-Tudela. 2002. Comparative evaluation of NCCLS M27-A and EUCAST broth microdilution procedures for antifungal susceptibility testing of Candida species. Antimicrob. Agents Chemother. 46:3644-3647.
Dannaoui, E., O. Lortholary, D. Raoux, M. E. Bougnoux, G. Galeazzi, C. Lawrence, D. Moissenet, I. Poilane, D. Hoinard, and F. Dromer. 2008. Comparative in vitro activities of caspofungin and micafungin, determined using the method of the European Committee on Antimicrobial Susceptibility Testing, against yeast isolates obtained in France in 2005-2006. Antimicrob. Agents Chemother. 52:778-781. doi:10.1128/AAC.01140-07.
Desnos-Ollivier, M., S. Bretagne, D. Raoux, D. Hoinard, F. Dromer, and E. Dannaoui. 2008. Mutations in the fks1 gene in Candida albicans, C. tropicalis, and C. krusei correlate with elevated caspofungin MICs uncovered in AM3 medium using the EUCAST method. Antimicrob. Agents Chemother. 52:3092-3098.
Desnos-Ollivier, M., F. Dromer, and E. Dannaoui. 2008. Detection of caspofungin resistance in Candida spp. by Etest. J. Clin. Microbiol. 46:2389-2392.
Forrest, G. N., E. Weekes, and J. K. Johnson. 2008. Increasing incidence of Candida parapsilosis candidemia with caspofungin usage. J. Infect. 56:126-129. doi:10.1016/j.jinf.2007.10.014.
Garcia-Effron, G., S. K. Katiyar, S. Park, T. D. Edlind, and D. S. Perlin. 2008. A naturally occurring proline-to-alanine amino acid change in Fks1p in Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis accounts for reduced echinocandin susceptibility. Antimicrob. Agents Chemother. 52:2305-2312. doi:10.1128/AAC.00262-08.
Garcia-Effron, G., S. Lee, S. Park, J. D. Cleary, and D. S. Perlin. 22 June 2009. Effect of Candida glabrata FKS1 and FKS2 mutations on echinocandin sensitivity and kinetics of 1,3-{beta}-d-glucan synthase: implication for the existing susceptibility breakpoint. Antimicrob. Agents Chemother. doi:10.1128/AAC.00443-09.
Garcia-Effron, G., S. Park, and D. S. Perlin. 2009. Correlating echinocandin MIC and kinetic inhibition of fks1 mutant glucan synthases for Candida albicans: implications for interpretive breakpoints. Antimicrob. Agents Chemother. 53:112-122. doi:10.1128/AAC.01162-08.
Garey, K. W., M. Rege, M. P. Pai, D. E. Mingo, K. J. Suda, R. S. Turpin, and D. T. Bearden. 2006. Time to initiation of fluconazole therapy impacts mortality in patients with candidemia: a multi-institutional study. Clin. Infect. Dis. 43:25-31.
Hajjeh, R. A., A. N. Sofair, L. H. Harrison, G. M. Lyon, B. A. Arthington-Skaggs, S. A. Mirza, M. Phelan, J. Morgan, W. Lee-Yang, M. A. Ciblak, L. E. Benjamin, L. T. Sanza, S. Huie, S. F. Yeo, M. E. Brandt, and D. W. Warnock. 2004. Incidence of bloodstream infections due to Candida species and in vitro susceptibilities of isolates collected from 1998 to 2000 in a population-based active surveillance program. J. Clin. Microbiol. 42:1519-1527.
Horn, D. L., D. Neofytos, E. J. Anaissie, J. A. Fishman, W. J. Steinbach, A. J. Olyaei, K. A. Marr, M. A. Pfaller, C. H. Chang, and K. M. Webster. 2009. Epidemiology and outcomes of candidemia in 2019 patients: data from the prospective antifungal therapy alliance registry. Clin. Infect. Dis. 48:1695-1703. doi:10.1086/599039.
Katiyar, S., M. Pfaller, and T. Edlind. 2006. Candida albicans and Candida glabrata clinical isolates exhibiting reduced echinocandin susceptibility. Antimicrob. Agents Chemother. 50:2892-2894.
Kuse, E. R., P. Chetchotisakd, C. A. da Cunha, M. Ruhnke, C. Barrios, D. Raghunadharao, J. S. Sekhon, A. Freire, V. Ramasubramanian, I. Demeyer, M. Nucci, A. Leelarasamee, F. Jacobs, J. Decruyenaere, D. Pittet, A. J. Ullmann, L. Ostrosky-Zeichner, O. Lortholary, S. Koblinger, H. Diekmann-Berndt, and O. A. Cornely. 2007. Micafungin versus liposomal amphotericin B for candidaemia and invasive candidosis: a phase III randomised double-blind trial. Lancet 369:1519-1527. doi:10.1016/S0140-6736(07)60605-9.
Laupland, K. B., D. B. Gregson, D. L. Church, T. Ross, and S. Elsayed. 2005. Invasive Candida species infections: a 5 year population-based assessment. J. Antimicrob. Chemother. 56:532-537.
Laverdiere, M., R. G. Lalonde, J. G. Baril, D. C. Sheppard, S. Park, and D. S. Perlin. 2006. Progressive loss of echinocandin activity following prolonged use for treatment of Candida albicans oesophagitis. J. Antimicrob. Chemother. 57:705-708.
Miller, C. D., B. W. Lomaestro, S. Park, and D. S. Perlin. 2006. Progressive esophagitis caused by Candida albicans with reduced susceptibility to caspofungin. Pharmacotherapy 26:877-880.
Mora-Duarte, J., R. Betts, C. Rotstein, A. L. Colombo, L. Thompson-Moya, J. Smietana, R. Lupinacci, C. Sable, N. Kartsonis, and J. Perfect. 2002. Comparison of caspofungin and amphotericin B for invasive candidiasis. N. Engl. J. Med. 347:2020-2029.
Odds, F. C., M. F. Hanson, A. D. Davidson, M. D. Jacobsen, P. Wright, J. A. Whyte, N. A. Gow, and B. L. Jones. 2007. One year prospective survey of Candida bloodstream infections in Scotland. J. Med. Microbiol. 56:1066-1075.
Ostrosky-Zeichner, L., J. H. Rex, P. G. Pappas, R. J. Hamill, R. A. Larsen, H. W. Horowitz, W. G. Powderly, N. Hyslop, C. A. Kauffman, J. Cleary, J. E. Mangino, and J. Lee. 2003. Antifungal susceptibility survey of 2,000 bloodstream Candida isolates in the United States. Antimicrob. Agents Chemother. 47:3149-3154.
Paderu, P., G. Garcia-Effron, S. Balashov, G. Delmas, S. Park, and D. S. Perlin. 2007. Serum differentially alters the antifungal properties of echinocandin drugs. Antimicrob. Agents Chemother. 51:2253-2256.
Pappas, P. G., C. A. Kauffman, D. Andes, D. K. Benjamin, Jr., T. F. Calandra, J. E. Edwards, Jr., S. G. Filler, J. F. Fisher, B. J. Kullberg, L. Ostrosky-Zeichner, A. C. Reboli, J. H. Rex, T. J. Walsh, and J. D. Sobel. 2009. Clinical practice guidelines for the management of candidiasis: 2009 update by the Infectious Diseases Society of America. Clin. Infect. Dis. 48:503-535. doi:10.1086/596757.
Pappas, P. G., C. M. Rotstein, R. F. Betts, M. Nucci, D. Talwar, J. J. De Waele, J. A. Vazquez, B. F. Dupont, D. L. Horn, L. Ostrosky-Zeichner, A. C. Reboli, B. Suh, R. Digumarti, C. Wu, L. L. Kovanda, L. J. Arnold, and D. N. Buell. 2007. Micafungin versus caspofungin for treatment of candidemia and other forms of invasive candidiasis. Clin. Infect. Dis. 45:883-893. doi:10.1086/520980.
Park, S., R. Kelly, J. N. Kahn, J. Robles, M. J. Hsu, E. Register, W. Li, V. Vyas, H. Fan, G. Abruzzo, A. Flattery, C. Gill, G. Chrebet, S. A. Parent, M. Kurtz, H. Teppler, C. M. Douglas, and D. S. Perlin. 2005. Specific substitutions in the echinocandin target Fks1p account for reduced susceptibility of rare laboratory and clinical Candida sp. isolates. Antimicrob. Agents Chemother. 49:3264-3273.
Pasquale, T., J. R. Tomada, M. Ghannoun, J. Dipersio, and H. Bonilla. 2008. Emergence of Candida tropicalis resistant to caspofungin. J. Antimicrob. Chemother. 61:219.
Perlin, D. S. 2007. Resistance to echinocandin-class antifungal drugs. Drug Resist. Updat. 10:121-130.
Pfaller, M. A., L. Boyken, R. J. Hollis, J. Kroeger, S. A. Messer, S. Tendolkar, and D. J. Diekema. 2008. In vitro susceptibility of invasive isolates of Candida spp. to anidulafungin, caspofungin, and micafungin: six years of global surveillance. J. Clin. Microbiol. 46:150-156.
Pfaller, M. A., and D. J. Diekema. 2007. Epidemiology of invasive candidiasis: a persistent public health problem. Clin. Microbiol. Rev. 20:133-163.
Pfaller, M. A., D. J. Diekema, L. Ostrosky-Zeichner, J. H. Rex, B. D. Alexander, D. Andes, S. D. Brown, V. Chaturvedi, M. A. Ghannoum, C. C. Knapp, D. J. Sheehan, and T. J. Walsh. 2008. Correlation of MIC with outcome for Candida species tested against caspofungin, anidulafungin, and micafungin: analysis and proposal for interpretive MIC breakpoints. J. Clin. Microbiol. 46:2620-2629. doi:10.1128/JCM.00566-08.
Reboli, A. C., C. Rotstein, P. G. Pappas, S. W. Chapman, D. H. Kett, D. Kumar, R. Betts, M. Wible, B. P. Goldstein, J. Schranz, D. S. Krause, and T. J. Walsh. 2007. Anidulafungin versus fluconazole for invasive candidiasis. N. Engl. J. Med. 356:2472-2482.
Rocha, E. M., G. Garcia-Effron, S. Park, and D. S. Perlin. 2007. A Ser678Pro substitution in Fks1p confers resistance to echinocandin drugs in Aspergillus fumigatus. Antimicrob. Agents Chemother. 51:4174-4176.
Rodriguez-Tudela, J. L., M. C. Arendrup, F. Barchiesi, J. Bille, E. Chryssanthou, M. Cuenca-Estrella, E. Dannaoui, D. W. Denning, J. P. Donnelly, F. Dromer, W. Fegeler, C. Lass-Florl, C. B. Moore, M. Richardson, P. Sandven, A. Velegraki, and P. E. Verweij. 2008. EUCAST definitive document EDef 7.1: method for the determination of broth dilution MICs of antifungal agents for fermentative yeasts. Clin. Microbiol. Infect. 14:398-405.
Rodriguez-Tudela, J. L., J. P. Donnelly, M. A. Pfaller, E. Chryssantou, P. Warn, D. W. Denning, A. Espinel-Ingroff, F. Barchiesi, and M. Cuenca-Estrella. 2007. Statistical analyses of correlation between fluconazole MICs for Candida spp. assessed by standard methods set forth by the European Committee on Antimicrobial Susceptibility Testing (E.Dis. 7.1) and CLSI (M27-A2). J. Clin. Microbiol. 45:109-111. doi:10.1128/JCM.01969-06.
Sipsas, N. V., R. E. Lewis, J. Tarrand, R. Hachem, K. V. Rolston, I. I. Raad, and D. P. Kontoyiannis. 2009. Candidemia in patients with hematologic malignancies in the era of new antifungal agents (2001-2007): stable incidence but changing epidemiology of a still frequently lethal infection. Cancer 115:4745-4752. doi:10.1002/cncr.24507.
Swinne, D., M. Watelle, C. Suetens, K. Mertens, P. A. Fonteyne, and N. Nolard. 2004. A one-year survey of candidemia in Belgium in 2002. Epidemiol. Infect. 132:1175-1180.
Thompson, G. R., III, N. P. Wiederhold, A. C. Vallor, N. C. Villareal, J. S. Lewis, and T. F. Patterson. 2008. Development of caspofungin resistance following prolonged therapy for invasive candidiasis secondary to Candida glabrata infection. Antimicrob. Agents Chemother. 52:3783-3785. doi:10.1128/AAC.00473-08.
Tortorano, A. M., J. Peman, H. Bernhardt, L. Klingspor, C. C. Kibbler, O. Faure, E. Biraghi, E. Canton, K. Zimmermann, S. Seaton, and R. Grillot. 2004. Epidemiology of candidaemia in Europe: results of 28-month European Confederation of Medical Mycology (ECMM) hospital-based surveillance study. Eur. J. Clin. Microbiol. Infect. Dis. 23:317-322.
Antimicrobial Agents and Chemotherapy Dec 2009, 54 (1) 426-439; DOI: 10.1128/AAC.01256-09
You are going to email the following Echinocandin Susceptibility Testing of Candida Species: Comparison of EUCAST EDef 7.1, CLSI M27-A3, Etest, Disk Diffusion, and Agar Dilution Methods with RPMI and IsoSensitest Media


Market Research Future (MRFR) projects the growth of the Global Industrial Alcohol Market to be steady and reach a significant valuation over the forecast period of 2017-2023. Industrial Alcohol experiences high demand from diverse industries such as automotive, chemical, pharmaceutical, and others since it has multiple advantages. Industrial Alcohol can be used as an antimicrobial agent, solvent and as a biofuel which supports the growth of the Industrial Alcohol Market. Industrial Alcohol is used for a broad spectrum of uses. Stringent emission norms and regulations mandating the use of ethyl alcohol as biofuel blend with petroleum fuels has spurred the growth of the Global Industrial Alcohol Market. Owing to raising awareness regarding greenhouse gas emissions, ethyl alcohol is gaining prominence as a biofuel as they are cleaner-burning fuels than diesel and pure gasoline. Industrial Alcohols are also used in the food & beverage industries too. In the pharmaceutical industry, Industrial Alcohol is used in antiseptics, drugs, lotions, and ointments while in the cosmetics industry it is used in perfumes, deodorants, and shampoos. Initiatives to develop low-cost biofuel is a trend which is likely to boost the growth of the market. Industrial Alcohol such as isopropyl alcohol is being widely used as a solvent in various industries. On the downside, fluctuations in the price of raw materials and complex approval process from regulatory bodies remain hurdles to the growth of the Industrial Alcohol Market.
The prominent players operating in the Global Industrial Alcohol Market include Cristalco (France), Exxon Mobil Corporation (U.S.), MGP (U.S.), Cargill, Incorporated (U.S.), Eastman Chemical Company (U.S.), BASF SE (Germany), Greenfield Global, Inc. (Canada), Raizen (Brazil), Royal Dutch Shell plc (Netherlands), and Green Plains Inc. (U.S.).
The Global Industrial Alcohol Market has been segmented based on Source, Type, and Application.
By Source, the market has been segmented into sugar & molasses, corn, grains, fossil fuels, and others.
By Type, the market has been segmented into ethyl alcohol, methyl alcohol, isopropyl alcohol, isobutyl alcohol, benzyl alcohol, and others. The ethyl alcohol segment is leading the market owing to its broad application in the gasoline blending process. Moreover, due to its antimicrobial properties, it is widely used across the cosmetic and pharmaceutical industry while the methyl alcohol segment is the second largest.
By Application, the market has been segmented into fuel, pharmaceuticals, personal care products, food ingredients, chemical intermediate & solvent, and others.
North America, Asia Pacific, Europe, Latin America, and the Middle East & Africa are the key markets for Industrial Alcohol. North America accounts for a substantial share of the Global Industrial Alcohol Market. The availability of raw materials makes the US the largest producer of ethanol as well as the largest consumer. The region has a huge market for pharmaceutical, and automotive industry and experiences inflated demand for industrial alcohol from these industries. Also, favorable schemes such as ethanol expansion in the US through the agricultural sector has contributed majorly to the North America Industrial Alcohol Market.
The Europe market is spurred by towering demand for Industrial Alcohol from the automotive, pharmaceutical, chemical, and other industries in the region.
Rapid industrialization in the Asia Pacific region has paved the way for the expansion of industrial alcohol market.
Latin America and the Middle East & Africa market are projected to exhibit considerable growth over the forecast period. The untapped markets in the region store ample opportunities for market expansion.


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Gelest promotes Jonathan Goff to VP and names Youlin Pan as Senior Research Fellow
Dr Jonathan Goff has been promoted to Vice President of Research & Development at Gelest, and Dr Youlin Pan has been named Senior Research Fellow, adding to his current title of Senior R&D Manager.
Dr Goff will broaden his role as a strategic partner in facilitating customer relationships, research initiatives and operational efficiencies. He will continue to manage the Polymer Development and Technical Services groups, and to help lead the Silanes & Metal-Organics and Research Engineering groups. An 8-year veteran of Gelest, Dr Goff has been an integral part of the Research & Development team's growth.
Dr Pan is Senior Research & Development Manager of Silanes & Metal Organics, and Senior Research Fellow. In addition to continuing to manage his research & development group, Dr Pan will broaden his role in developing and expanding the capabilities of Gelest Research & Development and overall operations. He has been with Gelest for over 21 years, and is a trusted resource for silicon and metal-organic chemistry expertise.
Related Materials Science news
Gelest Inc. Names Dr. Jonathan Goff as President
Perstorp further strengthens its specialty position within resins and coatings through acquisition of Di-Methylolpropionic Acid business from GEO
Indorama Ventures to expand packaging business into Vietnam strengthening market position in Asia-Pacific
The Heubach Group and SK Capital Partners Close Acquisition of Global Colorants Business of Clariant and Announce Appointment of Stefan Doboczky as Chief Executive Officer

redpajama_set_name: RedPajamaCommonCrawl


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Home » Organizing » Musicians Organize to Create Fair Streaming Agreements
Musicians Organize to Create Fair Streaming Agreements
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by Marc Sazer, President Recording Musicians Association and Member of Locals 47 (Los Angeles, CA) and 802 (New York City)
Even as our national government is mired in anti-labor policies and politics, the entertainment industry has experienced pioneering organizing over the past few years. Actors, writers, even video game workers, are organizing, acting, and gaining improvements. For musicians, our need to participate in the new streaming economy is an existential imperative. The last few years have seen our union negotiate key improvements from major media companies, especially in regards to our US pension fund, Music Performance Trust Fund, and Sound Recording Special Payments Fund.
But the rapid move across all media to streaming platforms threatens to leave musicians behind. This would be devastating, not just to those of us who work directly in recording studios, but for all of us. Our funds and our place in pattern bargaining would be damaged irrevocably, if we can't achieve fair compensation in streaming media. But perhaps the greatest harm would befall the next generation, and the next after them—aspirations dashed, a decent future denied.
We all have a stake in this, not just the house band players on the late night shows and the performers on The Voice and Dancing with the Stars. This includes musicians who prepare and perform the music on films, television, and streaming shows, from little indie and student projects to Star Wars, The Marvelous Mrs. Maisel, and The Orville; the players on records that are licensed into these media; as well as sideline musicians all over North America who bring these projects to life.
And we can do it! Take a look at some recent news headlines, as people working all across the entertainment industry seek real-life improvements through organizing.
"As Video Games Make Billions, the Workers Behind Them Say It's Time to Unionize"
Organizers with Game Workers Unite, a group that has sprung up in the last year to push for wall-to-wall unionization in the $43-billion game industry, kicked off each session with an icebreaker: "Damn the man." (Sam Dean, April 12, 2019 LATimes)
"Telemundo Actors Vote Overwhelmingly to Join SAG-AFTRA"
Actors at the Spanish-language TV network Telemundo have overwhelmingly voted to unionize with SAG-AFTRA, bringing to a close a protracted dispute between Hollywood's largest union and NBCUniversal, which owns the network. (David Ng, March 8, 2017 LATimes)
"SBS's La Ley Staffers Vote to Join SAG-AFTRA"
The on-air talent at Spanish Broadcasting System's Chicago radio station La Ley 107.9 (WLEY-FM) voted overwhelmingly to recognize SAG-AFTRA as their union.
(Veronica Villafañe, October 1, 2018, Mediamoves.com)
"Aaron Sorkin and David Chase Among the Writers Who Support WGA in Fight with Agents"
Ratcheting up the pressure in its fight with Hollywood talent agencies, the Writers Guild of America has released a statement of support from hundreds of its members who are saying that they intend to vote in favor this week of a new code of conduct that would limit agency practices, including the packaging of productions. (David Ng, March 25, 2019, LATimes)
"Actors' Equity Reaches Development Deal with Broadway Producers; Strike Over"
Actors' Equity Association and Broadway producers have reached what Equity is calling a "historic" agreement on a profit-sharing contract model for union members participating in early stage production development. The new lab agreement includes profit sharing, higher wages, and additional stage manager contracts, according to Equity. (Greg Evans, February 8, 2019, Deadline.com)
"Musicians Seek Streaming Residuals as Contract Talks Launch with Studios"
The AFM held a press conference Wednesday prior to the start of negotiations at the headquarters of the Alliance of Motion Picture and Television Producers in Sherman Oaks, Calif. Musicians are working under the terms of an extended contract that was signed in 2015 as a three-year deal. The AMPTP had no comment.
The AFM noted that SAG-AFTRA, the Writers Guild of America, and the Directors Guild of America have been able to negotiate residuals for films made for streaming, but musicians have been excluded. The union noted that musicians currently receive residual payments for secondary-market uses of theatrical and TV films, but not for films made for the Internet.
"As streaming consumption grows, the absence of streaming residuals will prevent musicians from being able to afford a home and feed their families, and threatens to erode the major contributions our members make to our local communities," said Ray Hair, AFM International President.
"AFM members must take on the changes in technology by ensuring that we maintain good jobs and a rightful place in the future of the industry," he added. "We are seeking a productive dialogue with AMPTP as we work to reach a fair resolution of these negotiations."
SAG-AFTRA President Gabrielle Carteris said in a letter, "Working people in the entertainment industry must face the changes in our business together. For generations, we have fought for quality jobs and won. Now, as the industry moves toward new media, we believe it is time to stand together again. Our members recognize the tremendous value that musicians bring to our films and television shows, and we support their demand for a fair contract for streaming." (Dave McNary, March 13, 2019, Variety)
Update on OCSM's 44th Annual Conference
OCSM's 44th Annual Conference will be held August 12-16 in Hamilton, OH, which will provide an overview of OCSM's part in the industry.
Solidarity Is the Heart and Soul of a Strong Union
The financial woes of our AFM Employer Pension Fund have created a serious challenge to our union solidarity.


This past week was full of lasts. Last theme training, last beach evangelism, last social, last Bible study training, last day of work, and last day at church. This morning, at the final Life Training, Mike Polley talked about staying awake to the gospel and spiritual realities when we go home.
The month of August isn't always easy for students. It lies right between intense community at Project and at school, and without that community, it can be easy to sleep all day or forget about getting time in the Word.
Mike said this is something to be aware of, and he went through four areas of caution: pleasure, comfort, personal performance, and emotions. Many of these require us to look inward and determine whether or not what happens at Project is because of God or because of the community. Performing for others comes from forgetting the gospel, thinking we can add things to the table.
"It's so dangerous because it just slips in," Mike said.
He also stressed how falling into this still just brings us back to the gospel – how it's these times of personal performance that we can fall back on the cross and know that we're forgiven.
Talking about emotions, Mike said that they should influence our decisions in light of truth. In the moments of lacking or negative emotions, we get to cling to Christ, because he is what's true.
Going in home in August doesn't mean we have a month to take a spiritual break. We should still do things out of a love and desire for God. But, if it's hard, that doesn't mean our faith has taken a huge step backward.
"You faith is perfected in Him and not what you do," Mike said.
After Life Training we spent time meditating on these things. Then, after church, many people went to Garden City Baptist Church for a church potluck, and many others went to Rioz, a Brazilian steakhouse. Tonight is our final D-group, and this week will be filled with more lasts as we continue to prepare for home.
Last evening, we held our Jobs Social. About 100 employees and employers came to enjoy fellowship with the students and hear a gospel message.
Everyone enjoyed light snacks and lemonade in the beginning. Then, students performed skits relating to their job responsibilities. For example, Wal-Mart student employees performed a skit in which they broke into dance and imitated their daily individial job duties. After the skits, Greg Morse, a student from University of St. Thomas, presented a gospel message.
By holding this social, we hope to minister to the local employees and employers that we work with. Students have been able to build relationships with their employers throughout the summer. Many have shared the Gospel with fellow employees. Our goal is that students would learn how to share their faith in the workplace. We also pray that fellow employees would come to know Jesus Christ as Lord.
C.S. Lewis said, "The very nature of joy makes nonsense of our common distinction between having and wanting." This is definitely a truth the Czech Republic team felt Sunday afternoon after spending five hours at Street Reach – a local homeless shelter and rehabilitation center.
This blog post is a little more subjective than what I've been doing. I just really want to tell you about the experience I had, which was similar to the other 20-some students who had the opportunity to volunteer.
Last week's prayer focus was the local community, and going off that, Hannah Mihalko was able to find a place we could serve this community.
Once we arrived at Street Reach, we spent no more than an hour completing the various cleaning tasks they had for us. But then, we ended up staying four more hours to talk, worship, and pray with people staying at and visiting the shelter.
Each person had a different story. Some were mentally challenged, some lived in the woods, some couldn't escape substance addictions, and some had families that seem beyond repair. I want to tell you about three people I talked to, and I would love for you to pray for them, even just once right now.
He said he has three kids from his ex-wife and one on the way from a different woman. He said he is a Christian, and he was able to articulate the gospel.
Pray that this would be true, and that Jesus would be so sweet to him. Pray that his brokenness would lead him straight to the cross.
In a way, he stuck out in the room. He's 23 years old (much younger than most there), and he appeared to be educated and friendly. Still, there is a lot of brokenness. He has a six-month old baby who's living with his ex-wife, who's in a homosexual relationship.
He did not seem to know Christ, so pray that God would work in his heart.
He was completely content. He said he'd been there for four or five days.
He said he's been able to share the gospel countless times, and he knows God has him there for a reason.
Pray that God would continue to do amazing things through Paul. Pray for his time at the shelter. I'm not sure if he'll be there for just a couple more days or a couple months. But God is good, so pray his strong and steady joy in Christ would not falter even a little.
I was so encouraged by all three of these conversations, as well as all the conversations everyone else was able to have. I've been amazed what doors God opens when my identity is completely in him and what he's done for me.
I – and the other students who went – were reminded about what matters. I think it's important to have perspective on that sometimes. Comfort, fashion, education – all these things rarely ever crossed the minds of the people at Street Reach. All people, from all places and all situations, have one thing in common: a desperate need for the gospel.
Last night, Justin Richardson led optional theology. He started the evening by making our purpose clear. "The point of this whole time is not vain discussions about theology. The aim is love."
The group started with about 10 or 15 people and dwindled down as the night progressed. Here are the questions asked with a brief summary of the responses. There are quite a few, so feel free to pick and choose what to read.
Dalen Miller: How are the festivals and the new moons and the Sabboth a shadow of the things to come?
The Old Testament law was very strict on these things. All of them are fulfilled in Jesus Christ. For example, the Sabboth is a day of rest, which is a shadow of the ultimate rest, which is found in Jesus.
Lucy Liu: How did Jesus, as one human, die as the sins for everyone?
Perfection is a key factor in how Jesus can cover our sins. In the Old Testament law, there were specific sacrifices for specific sins. In the New Testament, the only sacrifice is the death of one perfect human.
Rom. 5:12-18 talks about this.
Lucy Liu: Why didn't he go to hell, if he's taking our punishment?
He is not defeated by death; he was able to overcome death.
Ben Shepherd: How does the death of Christ cover people in the Old Testament? How were they saved, as they didn't believe in Jesus?
People in the Old Testament believed in God and trusted in his plan. God had a method for allowing people to believe in him.
Heb. 11:13-16 and 39-40 talk about people in the Old Testament being justified by faith.
People are not made perfect in the Old Testament. They are given yearly times with a priest and momentary sacrifices to atone for their sins. And throughout the Old Testament, we can see that all this is temporary.
The key was belief in God. How was that belief flushed out? Through their works – but ultimately, through faith.
Lisa Fitton: Is it possible that God exists outside of time and that the gospel did exist for them?
His death on the cross does extend to them in some way. For us, the basis for our salvation is based on belief in Jesus, so it's not exactly the same.
Leah Miller: What does that mean to delight yourself in the Lord and you will get the desire of your heart?
The verse is not talking about how we will get things like a new car if we just delight in Christ. It's talking about how when we delight in him, our desires change – improve – to be the desires God wants them to be. And he gives us those desires.
Anaite Castaneda: If everyone is created for a relationship Christ, why doesn't everyone have that?
Romans 9 could be a good place to go. While all human beings are created with hearts for knowing God, for some reason, God has chosen to harden some hearts. An example of this is Pharaoh.
AnnaLiese Durling: If, for people in the Old Testament, salvation was about belief in God, can people now be saved by just believing in God (and not the gospel)?
Rom. 1:18 talks about how people who don't know Jesus are without excuse. It's only through Jesus by which we can be saved. And all who God created to be saved will be saved.
Emma DeJong: In Jeremiah God tells Jeremiah not to pray for people. What's that about?
God poured out his wrath in a different way then than he does now, as it has all been taken by Jesus. Also, Jeremiah's motives in his prayers were more concerned for the people, rather than for God's glory.
AnnaLiese Durling: Did the Trinity exist before Christ?
In Gen. 1:2, we can see that in the very beginning of all creation, the Spirit was hovering over the face of the waters. The trinity always existed. The Son and the Spirit came into a manifestation of their role with the cross.
AnnaLiese Durling: I have a friend who's very moral. How do I talk about morality with him?
Christianity is not about becoming a better person. It's about saying, 'I'm OK.' James Lovallo offered that advice to remember that Christianity is not about works; it's about Christ. That's what he needs to hear the most.
Rachel Carlson: What does it mean to pray in faith?
God will always answer your prayers. There are three answers he gives: yes, no and maybe.
Rest in what God has done, and pray about that. Expect God to do big things. Also, pray what people prayed in the Bible.
AnnaLiese Durling: In Scripture memorization, how do you choose what to memorize?
It depends on where you are in the moment and where you are in Scripture memory. One of the purposes of memorizing Scripture is to know more about who God is.
At this point, the night became much more of an informal conversation. After a while Justin prayed for everyone in closure.
Now that the staff is gone, the team leaders are giving the messages. Justin Richardson led Bible study training yesterday morning. He discussed why we need to read the Bible and how our thoughts and emotions are involved.
"It is God's will for us to read His Word," Justin said.
But what about when we go through spiritual dry periods? Many people don't read their Bible unless they want to because they don't want to be legalistic. Justin addressed this, saying it is a misconception that obedience is primarily a result of emotion.
Emotions play a large role in the Christian life, but they are not a prerequisite for obedience. If somebody doesn't want to read their Bible, not reading would probably be their worst decision.
Justin also addressed the misconception that we are less pleasing to God when we are emotionally dry.
"Christian hedonism … is at the same time thrilling and crushing," he said.
We get glimpses of joy in Christ, but it can be crushing when we don't. However, we find that joy when we read the Bible – when we learn more about Christ. Justin said that "the Bible is a source for our emotions." He reminded everyone that the basis for God's pleasure in us is not because of us, but because of what Christ has done for us on the cross.
He continued to explain why we should read the Bible.
Proverbs 2:1-5: My son, if you receive my words and treasure up my commandments with you, making your ear attentive to wisdom and inclining your heard to understanding; yes, if you call out for insight and raise your voice for understanding, if you seek it like silver and search for it as for hidden treasures, then you will understand the fear of the Lord and find the knowledge of God.
The time has come for the project staff to return to their lives in Minnesota, Michigan and South Dakota, leaving Project to be completely student-run for the second month. Today and tomorrow all of the staff are packing and either driving or flying out.
Last night was the farewell party. To start things out, Jon Saunders, Megan Cogan and Elliot Stokes downed a box of ice cream – not faster than the team leaders, of course.
Then, the students from each campus prayed for their staff, and it's comforting to know that they are going to continue praying for us at Project.
If you want to pray for any of the 14 staff members from South Project, many of them will be raising support, so prayers would definitely be appreciated for that. Also, pray that the gospel would be greater and sweeter for them as they return, and that it would spread to their families, friends and students.
It's been a busy week since the last blog post.
We had a typical work day, with team time at 6 p.m. Each team (Goof Troop, Vector, and The Czech Republic) meets once a week to hang out and get to know each other better.
At 8 p.m., everyone returned to their rooms for D-groups – where each group of four or five students learn to love one another more through conversations, studying scripture, and praying.
Wednesday mornings student leaders and staff meet at Garden City Baptist Church for Leaders Training. Paul Poteat talked about the deep brokenness of everyone there, as well as everyone not there. He said that we need to remember that there are things deeper in the hearts of students than the things they do.
Then, the guys and girls from each team meets for their D-group. This is a refreshing time for room leaders and their team leader to catch up and offer one another encouragement and advice.
At 1:30 p.m. the whole project meets for Evangelism Training, followed by evangelism on the beach. This week, Jon Saunders talked about our fear of evangelism, and how our hearts show what we believe about the Father. He gave some practical reasons why we should not be scared of beach evangelism, but he also shared the real reason: God is a good god, and that he wants to give us good things.
So, we all went out in pairs and talked to people on the beach, asking them questions and sharing the gospel. Afterward, teams met to share stories about how it went, and then to pray about those specific people.
Wednesday night, many people went out on group dates (where a guys' room asks a girls' room on a date). At 8 p.m., Team Czech Republic provided s'mores for the week's encouragement. Overall, it was a relaxing evening.
Again, Thursday is a work day. But Thursday nights, we have our project social. Last night was the social of all socials.
enjoyed a feast of shrimp, potatoes, corn, and sausage – all piled on tables. Forks were unnecessary.
The night also included sing along with Paul. This was when Jordan Zurek's inner Taylor Swift came out, as well as when Ben Shepherd's Irish identity was confirmed.
Bobbing for limes was also a hit, but fun fact: Limes are cheaper than apples, but they sink. The night was finished with about three or four hours of dancing, even through down-pouring rain.
Then, some people relaxed and some went off to work.
To start our day, Mike Polley talked about money and possessions at Life Training. He exposed our selfishness when it comes to these things, and how we are prideful about how much or how little we have. Hearing the gospel was very freeing.
At 11 a.m., everyone went to church, and then afterward rested in a variety of ways. Later that night was an optional women's time, and the CCP group gave a presentation about India, explaining what they will be doing there and how we can pray for them.
This is the day the Lord has made! Pray that we will rejoice and be glad in it.
For most of his message, Matt talked about how good the Good News is.
Hebrews 10:14: For by a single offering he has perfected for all time those who are being sanctified.
But how can we hear this and so often feel nothing? How can there be such numbness to the gospel in our hearts?
Matt brought up three sedatives that put us to sleep when it comes to the gospel.
One is a shallow understanding of sin. This blog post could never fully describe the weight that was felt in the fellowship hall of Garden City Baptist last night. With full conviction, holding nothing back, Matt preached about sin that was in every heart in the room.
We must know we are broken to understand the gospel.
Another sedative is the distraction of personal performance. This one hit home for many people (probably everyone). He talked about how we perform for people, and we perform for God. He stressed how there is nothing we can do for God.
Luke 18:9-14 is a perfect passage for what we have a right to say before the Lord. "God, be merciful to me, a sinner!" We can't pay God back with obedience or gratitude, and there is nothing we can do to improve him or improve ourselves.
But, this is such good news!
1 Cor. 3: 21-23: So let no one boast in men. For all things are yours, whether Paul or Apollos or Cephas or the world or life or death or the present or the future—all are yours, and you are Christ's, and Christ is God's.
All work has been done. It is finished. Now, we can rejoice that all is our's because we are Christ's.
The third and final sedative is the forgotten reality of eternity. We are now free to say that God's wrath has been taken away from me forever, and I will never see it.
…It's off to work we go.
A majority of the students on Project have been blessed with full-time jobs in the community. Over 50 work at Walmart. Also, there are some at Chic-Filet, Krystal Burger, McDonalds, Sonic, Buy Low, KFC, Painters, and a few various internships.
For many students, Saturdays mean longer hours, as there are no scheduled Project events.
Tonight, people are relaxing from a long day flipping burgers and stocking shelves. Some people are playing pool ball, some are throwing a frisbee, some are playing cards, and some are plucking the ukulele. As Rachel Sandager, Caleb Nordquist, Lucy Lou, and Max Cruz play Egyptian Rat Race (a card game), they reflect on their day.
Awake to the Scriptures: What is the Bible?
This morning everyone walked across the parking lot for Bible Study Training at Garden City Chapel.
Elliot Stokes went through what the Bible is and why we read it.
"Here are the most important things about the universe … and you're being walked through those things (when you read the Bible)," he said.
Elliot started by explaining that all of the Scriptures are about Jesus.
He stressed how the Bible is God's spoken word – that God literally speaks things into existence. He creates and defines truth by his words. He referenced 2 Timothy 3:15-16 and 2 Peter 3:15-16 to explain this.
Elliot called everyone to open the Bible realizing that they are hearing from God.
He also talked about how "the Bible tells you who God is, and people don't like that." People like to make up things about God and choose what they like or don't like to believe.
"The real God is way, way, way better than anything you would make up," Elliot said.
After the message, some of the students said this was convicting, reflecting on how it can be easy to seek forgiveness by getting in the Word rather than falling before the cross.
Bible Study Training is a weekly event. Every Friday morning, a different message is presented, and then we practice the inductive method of reading the Scriptures. This morning, we read today's verse from Colossians: Col. 1:24.
Now I rejoice in my sufferings for your sake, and in my flesh I am filling up what is lacking in Christ's afflictions for the sake of his body, that is, the church.
This week at project we are focusing on the topic of relationships. What does it mean to have a God-honoring and Christ-centered relationship? This was a central question that was highly focused on.
Andrew and Lauren Holbrook gave the relationships seminar. Andrew Holbrook is the regional director of Campus Outreach Charlotte. In their talks, they highlighted the fact that relationships are to point us more to Jesus Christ, not merely another person. In addition, they walked through how biblical marriages symbolize Christ's love for His church. They also answered many misconceptions our culture has towards relationships.
Many students were encouraged by these talks. There was much conversation by the students evaluating their relationships and setting goals and guidelines for healthy relationship. Remya Thomas, an international recent graduate from St. Stephens in Delhi, India, was quoted saying, "It's not all about marriage, and It's not about the intimacy we share with a man or a woman, but marriage is a glimpse of the intimacy we will share with the Lord." This thought was what stuck out to her the most from relatiosnhips week.
outside and prayed together for the people of Dhobi, India. They prayed that these people would hear the gospel of Christ. Remya was able to provide us with insight into the people.
Project was incredibly blessed by the Holbrooks coming to speak about relationships. Students are excited to grow in godly relationships with one another and in dating relationships. We are also delighted that Remya is with us this summer. Her being here is a continual reminder to pray for the nations that have not yet been reached with the gospel.
Matt Reagan said that a couple times last night as a part of his response to the questions at Theology Time.
Every Sunday night, a couple people on staff answer any questions students may have about God, Christianity, and anything else. Last week, Matt Reagan and Jon Saunders and about 20 students huddled under the tent during a massive thunderstorm to have this discussion.
Three nights ago, Paul Poteat and Matt Reagan answered a variety of questions, from heaven and hell to angels and demons to sin and salvation. The format of the discussion is question-and-answer. Students ask questions, and Paul and Matt point to places all over the Bible for answers.
They are not the layers found in Dante's Inferno, but the idea of levels in heaven is Biblical. 1 Cor. 3: 9-17 talks about this.
Matt said that Not extra possessions in heaven.
In 1 Thes. 2, the apostle Paul talks about how the people he is writing to are his "glory and joy" at the coming of Jesus. Matt stressed that this language of boasting and reward ultimately gives Jesus all the glory.
There will be rewards in heaven. Matt said, "You ought not worry too much about your heavenly rewards." He stressed that there will be no envy. The way we will think about our own rewards and other rewards will be in a completely sinless way.
Paul added that rewards don't come when we think intentionally about the rewards. It's the gospel that produces good fruit. "Focus on who Jesus is for you and not who you are for Jesus," Matt said.
They also pointed out that Luke 12:47-48 talks about levels of Hell.
Matt and Paul brought up Hebrews 7-9, suggesting that Ben look there to understand how the sacrificial system is insufficient.
At the end of the discussion, Paul added a reference to Rom. 8:5-7, explaining how the flesh is hostile to God.
Paul said that one of his past professors told him that of all the ethical things we deal with, God is number one and angels are number two. Paul also brought up 2 Kings 6:16, where Elisha had faith in angels during battle. He added that people in America are deluded that supernatural forces are not actually real.
Matt said that there is a whole other realm of cosmic powers that we just don't see – that angels and demons are at war. He mentioned Daniel 10 as an example of this.
Paul brought up how in 1 Cor. 11:10, the language shows how the people of that time knew about angels – more than we do now.
• What does it mean when we say that Jesus was begotton?
• What does the Bible say about levels of heaven?
• Is Hell God turning away from you in rejection or to you in anger?
• Is it possible for people to fall from Heaven (like Satan)?
• In Colossians 1, what is the kingdom of his Beloved Son?
• Would it have been just for God not to save anyone?
• If someone were to ask, "Why can we not atone for our own sin," where do I point them?
• What do Jews believe about when the Messiah comes?
• How did Adam and Eve populate the world?
• What exactly are angels? How do they interact with humans?
• How do demons fit into us being accountable for our sins?
• Where is the name Lucifer in the Bible?
• What's the difference between a new earth and heaven?
• My roommate thinks she can become God by being in nature more. What can I say to her?
*Including the responses to each one of these questions would make this post longer than a daily blog post should be. This might also be a good time to ask for forgiveness for any poor note taking on my part. If I include exact words from anyone, I will put them in quotations. Otherwise, assume that I am paraphrasing.
Five-cent coffee is just around the corner. But at the 2011 Summer Training Project, caffeine has nothing to do with staying awake.
Awake is the theme for the summer, and Elliot Stokes gave the first theme training June 2. He said there is an awake way to live and a sleepy way to live. We want to be awake to all spiritual truth, and we want to see the world through a Biblical lens. This is what we will be focusing on for the two months in Garden City, S.C.
So what's been happing at Project for the past two weeks?
The staff and student leaders arrived late-afternoon May 30 and were instantly hard at work. In a matter of a couple days, 37 rooms that hadn't been unlocked since last summer were made livable.
On June 2, the rest of Project arrived. As each new car unloaded, Larry Martini's energy level kept rising. After the 22-hour car ride, the disciples were greeted with lots of cheering, Jordan Zurek sprawled on top of their car, and a popsicle.
It hasn't taken long for the room leaders and disciples to get to know one another. Each room has one leader and three or four disciples. In each relatively small space, friendships have been formed, meals have been made, sin has been exposed, and dance parties have happened.
Students have also grown through learning about evangelism. Jon Saunders challenged people by saying that we are natural evangelists for what we love most. Each Wednesday, everyone at Project goes out on the beach and shares the gospel with people. On June 8, people paired off and evangelized for a little over an hour. Then, everyone sat around the pool and shared how their discussions went, praying about specific individuals.
Mondays, Tuesdays, Thursdays, Fridays, and Saturdays are work days. Students work at a variety of places, including Walmart, McDonalds, Krystal Burger, Sonic, and Buy Low. The Lord was so gracious to provide every person a job, and we are working to be thankful as well as joyful about work.
Every Thursday night, we have a project social. We have had two so far. For the first, everyone went to see Kung Fu Panda II, and all the disciples dressed their room leaders up as a jungle animal. It was pretty interesting. For the second social, we had a "boat social." To sum it up: there were three spectacular performances by the pool – one with pilgrims, one with pirates, and one with The Little Mermaid. Each team had to build a boat that could take people across the pool, all the while acting out the scene.
On Sunday mornings, each team attends a different church that is close to Project. Sunday nights, there is optional worship and optional theology time. For many people, Sundays are a day to rest.
Hello! I'm very excited to be blogging this summer about STP. This first post is an extremely brief summary of what has happened so far. Somebody could probably right a novel about each day, but unfortunately, our schedule is pretty packed.
I'm hoping to post at least once every day. As of right now, I'm planning on posting each morning about something that happened the previous day. So basically, stay tuned and be ready to hear all about Project!
Students, bring your laptops to the prep party tomorrow to register for jobs. This is an email reminder that you are having a MANDATORY Informational/prep party for the Summer Training Project on Friday, May 6 at 6pm at Bethlehem Baptist Church North Campus. If you need a ride, contact a staff member or an older student that will be attending the party.
Bank Accounts: •	Every summer we strongly encourage students to sign up for a local bank account in Myrtle Beach, SC •	We can't force you to sign up for an account, but we dont want you to have to pay $10 fee every time you need to cash a check •	It generally works very well to have an account in Myrtle Beach before you start your job at project •	If you have a wells fargo account, that now works with Wachovia We are providing you the resource tomorrow at the Prep Party to get your account setup before we get to project so you can have your debit/check cards established when you arrive.


Wow! Looks like you are into gas forges big time. If you haven't done so, I recommend you swing over to www.iforgeiron.com blacksmithing forum and check out the info on gas forges. There is a ton of info there and lots of guys who are very knowledgeable about gassers. Sign up and join the group, you won't regret it. Be sure to put your location, of some kind, in the signup or the boo-birds will hound you...LOL. There is a section devoted strictly to gas forges, so spend some time reading through it as well.
Great post Mike, thanks for taking the time. I'll be trying to make one of these before long, and your info should be real helpful.
Another good site for info is the Blacksmithing for Beginners page on Facebook. The discussion is kind of painful, but there's some good info in the archive pages.


The winner of the John Parfitt Trophy for the club member completing the most miles in Sportives goes to Heather Mackay for the second year running. Heather completed a magnificent 5880 miles in Audax and sportives despite missing part of the early part of the season while recovering from a foot operation. Well done Heather.
In second place was David Dyer who completed 1053 miles and the bronze medal went to Sarah Davies who completed 877 miles. Well done to both of you. Well deserved with all the contribution you both make to our Club activities and particularly the Clock Tower club run.


text:
Mike Aviles' patience paves way for win
By Bill Ballou TELEGRAM & GAZETTE STAFF
Jun 27, 2012 at 12:01 AM Jun 27, 2012 at 1:38 PM
It was not quite a five-run homer, or a 26-pitch perfect game, but what Mike Aviles did in the seventh inning last night ranks as up there with other Tales of the Unexpected.
He walked.
When Aviles walked, he loaded the bases for Dustin Pedroia, who followed with a two-run single that provided Boston with the go-ahead runs in a 5-1 victory over the Blue Jays.
The Red Sox are 10-3 in their last 13 games.
"That was a big play," manager Bobby Valentine said of the base on balls. "Those are not his forte, if you hadn't noticed."
In 48 games since May 4, Aviles has as many home runs as walks — four.
"That was pretty fun," he said. "I was looking for a pitch I could drive to the outfield and took two terrible swings."
David Pauley, the former Red Sox pitcher, was on the mound for Toronto and being a sinker-slider guy, Aviles knew that if he stayed disciplined and didn't swing at anything belt high or lower, the balls would be out of the strike zone.
He was right, and Pedroia shortly thereafter delivered the winning runs.
For a long time, it looked like Boston would waste the best effort of the year by Daisuke Matsuzaka, who gave up a run in the first, then pitched shutout baseball. The problem was that Blue Jays starter Aaron Laffey didn't give up any runs in the first and pitched shutout baseball, so it was a 1-0 game going into the bottom of the seventh.
With Jason Frasor in to relieve Laffey, Jarrod Saltalamacchia came up with two out and hit another clutch homer, this one into the Monster seats. That tied it up and the rally continued, with Pedroia finishing it off.
It was Saltalamacchia's 14th homer this season and 30th as a Red Sox.
"He is a confident player," Valentine said of his catcher. "Some things that might have hampered him in the past seem to be gone."
The winning pitcher was Andrew Miller, who faced one batter.
For Matsuzaka, this seems to be payback for his 2008 season, when he was 18-3 and every thing that could have gone right for him did. So far in his four starts, the Red Sox have scored three runs during the 21-2/3 innings he has been in the game.
Statistically, this was Matsuzaka's best performance since his return from Tommy John surgery. He allowed just the one run in 5-2/3 innings, walked one, fanned five and gave up six hits. His ERA dropped from 6.06 to 4.98
However, once again the first inning was his worst inning, although his teammates did not help much.
Matsuzaka, who gave up three runs in the first on June 21 and two on June 15, allowed a leadoff double to Brett Lawrie. Colby Rasmus then grounded to short and Lawrie broke for third; Mike Aviles threw there ahead of him, but Lawrie slid in safely.
After Jose Bautista fouled out, Lawrie scored on Edwin Encarnacion's infield hit to third.
"To give up just one run in that inning is good pitching," Valentine said. "That was his best outing yet."
Toronto had a couple of chances to extend its lead before Boston mounted its rally. One reason the Jays did not was a splendid play by Aviles to end the Toronto sixth. It had runners at first and third and speedster Rajai Davis up with two out.
Davis hit one into the hole and Aviles got it on the backhand, then fired quickly to second to force Yunel Escobar and end the inning.
"That was a play I had to make to get us out of that inning and into the dugout," Aviles said. "With (Davis) running, I knew if I went to first with it, he'd be safe."
After they got three in the seventh to take the lead, the Sox added two more in the eighth on doubles by David Ortiz and Adrian Gonzalez and a single by Cody Ross.
So, Boston wound up with five runs in large part because Aviles took one walk.


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'Extending economic models to include environmental emissions and environmental policy is now commonplace. Yet, the linking is often ad hoc and based on peculiar assumptions. In this book, Rob Dellink shows how emissions and emission reduction policies for five different environmental issues can be included in a forward-looking computable general equilibrium model, building emissions into production and consumption functions. At the technical level the book is superb and stands out for its exact specification of how this is done.' - Richard S.J. Tol, Hamburg University, Germany, Free University Amsterdam, The Netherlands and Carnegie Mellon University, US This important new book enhances our understanding of the dynamic interactions between economic activity, economic growth, pollution abatement and environmental policy. It addresses one main policy problem: how can the direct and indirect costs of environmental policy for multiple pollutants be properly assessed in an applied model?
China's Dilemma - Economic Growth, the Environment and Climate Change examines the challenges China will have to confront in order to maintain rapid growth while coping with the global financial turbulence, some rising socially destabilising tensions such as income inequality, an over-exploited environment and the long-term pressures of global warming. China's Dilemma discusses key questions that will have an impact on China's growth path and offers some in-depth analyses as to how China could confront these challenges. The authors address the effect of the global credit crunch and financial shocks on China's economic growth; China's contribution to greenhouse gas emissions and emissions reduction schemes; the environmental consequences of foreign direct investment in China; the relationship between air pollution and mortality; the effect of climate change on agricultural output; the coal industry's compliance with tougher regulations; and the constraints water shortages may impose on China's economy. It also emphasises the importance of managing the rising demand for energy to moderate oil price increases and placating domestic and international concerns about global warming. In the thirty years since China started on the path of reform, it has emerged as one of the largest and most dynamic economies in the world. This carries with it the responsibility to balance the requirements of key industries that are driving its development with the need to ensure that its growth is both equitable and sustainable. China's Dilemma highlights key lessons learned from the past thirty years of reform in order to pave the way for balanced and sustained growth in the future.
Climate change and energy consumption are at the forefront of current environmental debate. While energy is essential to the functioning and survival of our societies, the environmental impact that energy consumption is having, particularly on climate change, is a growing concern and the design and practicalities of energy and energy-related environmental policies are under constant scrutiny. This innovative new book not only addresses the economic assessment of environmental and energy policies but also discusses the efficiency and distributional consequences these policies have for producers and consumers. With contributions from leading academics in the field, this comprehensive volume uses a variety of methodological approaches with which to explore a number of pertinent issues, including several studies on the EU Emission Trading System, as well as more advanced topics such as indeterminacy and optimal environmental public policies, energy-saving technological progress, oil shocks and energy transitions and policy design. Combining theoretical and empirical work, this timely book is a significant contribution to the existing literature and deals with issues at the frontier of current economic knowledge. Economic Modelling of Climate Change and Energy Policies is a unique and informative book and will have widespread appeal amongst scholars, students and policymakers.


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Smithland is een plaats (city) in de Amerikaanse staat Kentucky, en valt bestuurlijk gezien onder Livingston County.

Demografie
Bij de volkstelling in 2000 werd het aantal inwoners vastgesteld op 401.
In 2006 is het aantal inwoners door het United States Census Bureau geschat op 396, een daling van 5 (-1,2%).

Geografie
Volgens het United States Census Bureau beslaat de plaats een oppervlakte van
1,6 km², geheel bestaande uit land. Smithland ligt op ongeveer 100 m boven zeeniveau.

Plaatsen in de nabije omgeving
De onderstaande figuur toont nabijgelegen plaatsen in een straal van 20 km rond Smithland.

Externe link

Plaats in Kentucky

redpajama_set_name: RedPajamaWikipedia