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A fluorescence in situ hybridization (FISH) technique based on binding of a rhodamine-labelled oligonucleotide probe to 16S rRNA was used to estimate the numbers of ribosome-rich bacteria in soil samples. Such bacteria, which have high cellular rRNA contents, were assumed to be active (and growing) in the soil. Hybridization to an rRNA probe, EUB338, for the domain Bacteria was performed with a soil slurry, and this was followed by collection of the bacteria by membrane filtration (pore size, 0.2 μm). A nonsense probe, NONEUB338 (which has a nucleotide sequence complementary to the nucleotide sequence of probe EUB338), was used as a control for nonspecific staining. Counting and size classification into groups of small, medium, and large bacteria were performed by fluorescence microscopy. To compensate for a difference in the relative staining intensities of the probes and for binding by the rhodamine part of the probe, control experiments in which excess unlabelled probe was added were performed. This resulted in lower counts with EUB338 but not with NONEUB338, indicating that nonspecific staining was due to binding of rhodamine to the bacteria. A value of 4.8 × 108 active bacteria per g of dry soil was obtained for bulk soil incubated for 2 days with 0.3% glucose. In comparison, a value of 3.8 × 108 active bacteria per g of dry soil was obtained for soil which had been air dried and subsequently rewetted. In both soils, the majority (68 to 77%) of actively growing bacteria were members of the smallest size class (cell width, 0.25 to 0.5 μm), but the active (and growing) bacteria still represented only approximately 5% of the total bacterial population determined by DAPI (4′,6-diamidino-2-phenylindole) staining. The FISH technique in which slurry hybridization is used holds great promise for use with phylogenetic probes and for automatic counting of soil bacteria.
For a long time plate counting has been inadequate for estimating the active populations of bacteria in soils (8, 40), and numerous methods have been tested as alternatives. Recently, the frequency of dividing cells (9) and counts of 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride-reducing cells (35), 5-cyano-2,3-ditolyl tetrazolium chloride (CTC)-reducing cells (48), and [3H]thymidine- or [14C]leucine-incorporating cells (3, 15) have been used to determine active bacterial populations. The limitations of these assays include inaccurate conversion factors in the frequency-of-dividing-cell, thymidine, and leucine methods (3, 9, 13, 17), as well as difficulties in using the CTC and thymidine methods with certain soils (15, 48).
The fluorescence in situ hybridization (FISH) method developed recently has great potential for improving determinations of active populations of soil bacteria (2, 14, 20, 42, 49). Oligonucleotide probes are designed based on signature nucleotide positions in the bacterial 16S rRNA and may be used to target either a narrow group of organisms or a broad group of organisms. High levels of rRNA (equivalent to high ribosome numbers), which result in high detection signals with the FISH technique, are observed in active populations of bacteria in which protein synthesis is occurring either in nondividing or dividing cells (12, 30, 37). In contrast, starved bacteria are known to contain low levels of rRNA (18) but might be detected by FISH under some circumstances (44). Similarly, 16S rRNA levels decrease below the detection level in stressed bacteria (44), and moribund bacteria are not expected to contain detectable levels of rRNA.
The majority of soil bacteria are known to be less than 0.5 μm in diameter (6, 7, 8, 17, 27), and it has been suggested that this population of small bacteria consists mainly of gram-positive organisms (32). These bacteria do not grow well under laboratory conditions (7), and their growth rate, as determined by the thymidine technique, may be lower than the growth rate of larger bacteria (4). Still, the polulation dynamics and functional role of the small bacteria are unclear (45). With the FISH technique it may be possible to directly detect the active bacterial populations in the soil and to determine the size class distribution and the responses of the bacteria to different soil conditions.
FISH protocols for bacterial cells that are extracted from soil (20) and bacterial cells that occur in smears (49) are difficult to perform because bacteria associated with soil particles are excluded and because the background fluorescence signals from the soil smears are high, respectively. To determine bacterial numbers in soil without an extraction step, a slurry hybridization protocol followed by membrane filtration and fluorescence microscopy was developed (14). By this procedure, it is possible to include the important fraction of adsorbed bacteria reported to exhibit a high level of activity (5) and to exclude the high background signal from minute clay and organic soil particles by collecting bacterium-sized particles with a filter. It should be noted that the slurry hybridization-membrane filtration method was originally developed with laboratory-grown bacteria added to soil. Hence, when indigenous soil bacteria are counted, some limitations due to nonspecific staining may be encountered. The aims of this investigation were, therefore, to determine the populations of active bacteria belonging to different size groups in soil and to modify the slurry hybridization protocol to compensate for nonspecific staining in soil samples.
Soil sampling and treatments.Bulk soil samples were collected from a sandy loam soil (63% sand, 17% silt, 17% clay, 1.1% C, pH [CaCl2] 5.5 [26]) at an experimental farm (Højbakkegård, Tåstrup, Denmark). This freshly collected soil was used during the developmental study to modify the slurry hybridization protocol. In additional experiments, glucose was added or a drying-rewetting cycle was included as described below before the bacterial populations were counted. All of the soil samples used for hybridization (5 to 10 g) were fixed in 100 ml of an ice-cold (pH 7.0-buffered) paraformaldehyde solution (1). After fixation overnight at 4°C, each sample was homogenized three times for 1 min in a Waring blender operated at the maximum speed. The cup containing blended soil was cooled on ice for 3 min between homogenization steps. The blended sample was stored at −20°C after 140 ml of filter-sterilized distilled water, 10 ml of 1 M Tris-HCl buffer (pH 7.5), and 250 ml of 96% ethanol were added. The dry weight of the soil sample was calculated on the basis of the soil moisture content.
Total counts of soil bacteria as determined by DAPI staining.DAPI (4′,6-diamidino-2-phenylindole) staining was used to determine the total numbers of soil bacteria. Samples corresponding to 0.1 mg (dry weight) were stained in 10 ml of filter-sterilized distilled water containing 1 μg of DAPIs per ml for 10 min (14, 36). Bacteria were collected by filtration through a 0.2-μm-pore-size black polycarbonate filter (diameter, 25 mm; Millipore). The filters were mounted in paraffin oil, and counts were determined with a fluorescence microscope as described below for the filter specimens obtained from in situ slurry hybridization experiments.
In situ slurry hybridization with rhodamine-labelled oligonucleotide probes.To target the active soil bacteria, 16S rRNA probe EUB338 (for the domain Bacteria) having the nucleotide sequence 5′-TGAGGATGCCCTCCGTCG-3′ (43) was used. A different probe, NONEUB338, having the nucleotide sequence complementary to the nucleotide sequence of EUB338, was used to determine the nonspecific binding of a probe to the soil bacteria. The nucleotide probes were labelled with rhodamine (tetramethyl rhodamine isothiocyanate) (Sigma Chemical Co., St. Louis, Mo.) at a 1:1 ratio by using an amino linker (Applied Biosystems, Foster City, Calif.), and the rhodamine-labelled probes were purified by high-performance liquid chromatography. The staining intensities of different batches of the probes varied slightly because of slight differences in the ratio of rhodamine to oligonucleotide (data not shown).
Sample volumes equivalent to 0.5 mg (dry weight) of soil were taken directly from the blended samples while they were being stirred magnetically. A dehydration process in which a sample was centrifuged (10,000 × g , 5 min) and then resuspended in 80 and 100% ethanol was used (14). Soil pellets were then vacuum dried to evaporate the ethanol and resuspended in 200 μl of standard hybridization buffer (0.9 M NaCl, 20 mM Tris [pH 7.0]) containing 0.1% Nonidet P-40 detergent (Sigma). The samples were centrifuged again, and each pellet was resuspended in 200 μl of hybridization buffer containing 15% formamide and incubated at 37°C for 30 min. The effects of formamide concentrations ranging from 0 to 30% on the cell counts were determined during the developmental study.
Then rhodamine-labelled probe EUB338 or NONEUB338 was added to each slurry at a concentration of 12 ng μl−1. This concentration was found to result in optimal staining, and higher probe concentrations did not result in higher cell numbers (data not shown). The standard protocol developed also included adding unlabelled probes EUB338 and NONEUB338 at a concentration of 36 ng μl−1together with rhodamine-labelled probes EUB338 and NONEUB338, respectively. The effects of concentrations of unlabelled probe ranging from 0 to 36 ng μl−1 on the cells counts were determined during the developmental study.
Hybridization took place at 37°C for 18 h by using continuous rotation in a hybridization oven. In comparison, a shorter hybridization time (3 h) resulted in lower counts of indigenous bacteria (the counts were reduced 50% or more) (data not shown). After hybridization each slurry was transferred to 10 ml of hybridization buffer containing 15% formamide and incubated at 37°C for 10 min. The significance of this washing procedure was determined during the developmental study by using wash times ranging from 2 to 60 min.
After hybridization, the soil bacteria were collected by filtration onto 0.2-μm-pore-size black polycarbonate filters (diameter, 25 mm; Millipore) in a filtration manifold with stainless steel chimneys which were prewarmed to the hybridization temperature. The filters were rinsed with 20 ml of filter-sterilized distilled water at room temperature (25°C) and stored at 4°C until fluorescence microscopy counting was performed (within 1 week). For longer periods of storage, the filters were frozen at −20°C. Prior to microscopy, the filters were mounted on glass slides in liquid paraffin oil.
Fluorescence microscopy counting and determining the size classes of active soil bacteria.To count the bacteria in different size classes, a Zeiss Axioplan 2 fluorescence microscope equipped with a Plan-Neofluar objective (magnification, ×100; numerical aperture, 1.3) and a 100-W type HBO high-pressure mercury lamp was used. Zeiss no. 2 and 15 filters were used to visualize bacteria stained with DAPI and rhodamine, respectively. Each bacterium was assigned to one of the following three size classes as defined by spheres of the G12 New Porton graticule (27, 34): small, medium, and large, representing cell widths of 0.25 to 0.5, 0.5 to 0.7, and 0.7 to 1.0 μm, respectively. This classification was chosen since bacteria belonging to the same species predominantly change in cell length rather than in cell width in response to variable growth conditions (24).
For soil samples hybridized with a rhodamine-labelled probe, autofluorescence may sometimes give erroneous results (14, 28). Therefore, a bacterial cell was counted only if autofluorescence was absent, which was ascertained by placing a Zeiss no. 2 filter (normally used for DAPI staining with UV light) immediately behind the rhodamine filter without moving the microscopic field. Autofluorescing cells were never detected with the Zeiss no. 15 filter when samples were prepared for microscopy without the rhodamine probe.
During the developmental study, at least 200 fields of 50 by 50 square units of the G12 New Porton graticule were inspected for each filter. In subsequent studies of treated soils, at least four replicates of hybridization experiments were used, and at least 100 cells were counted for each sample. At low cell densities a total of 600 microscopic fields were inspected, which corresponded to 1% of the sample filter. Statistical differences between counts obtained with different treatments were analyzed by a d test (10). The d test was used because the variance was not the same for all of the data sets compared. Comparable levels of variance are required to perform the t test. Thed test follows a Fisher-Behrens distribution, as described by Campbell (10).
For selected samples confocal laser scanning microscopy (CLSM) was used to confirm the distinction of active bacteria from nonactive bacteria or nonbacterial soil particles. The type of confocal microscope used (model TCS4d; Leica Laser Technik, Heidelberg, Germany) and the associated equipment have been described previously by Hansen et al. (23). In this study, we used soil samples pretreated with glucose to stimulate bacterial activity. The filters were first scanned at random by using the optical part of the confocal microscope, and active bacteria were separated from nonactive bacteria and nonbacterial soil particles by using the following criteria: light intensity, morphology, and color. Particles were confirmed to be active bacteria by examining pictures taken by CLSM. Each recording consisted of a stack of images with a vertical distance of 0.3 μm. The stack was subsequently combined into one image by maximum intensity projection performed by the software of the model TCS4d microscope.
Optimization of hybridization conditions.To count indigenous, active soil bacteria, the protocol for in situ slurry hybridization (14) had to be optimized, notably because nonspecific staining was expected to be significant. Addition of formamide to the hybridization buffer is known to decrease the melting point of the DNA double helix (11), and the optimal formamide concentration for hybridization at 37°C was determined to obtain a high-stringency protocol. Figure1 shows the effects of different formamide concentrations on the hybridization signal for both specific probe EUB338 and nonspecific probe NONEUB338. The total population of cells generally gave the highest counts in the presence of 15% formamide for both probe EUB338 and probe NONEUB338, and this standard concentration (15% formamide in hybridization buffer) was used for all subsequent experiments. All three size classes of bacteria exhibited the same pattern as the total bacterial population (data not shown). Hence, significantly higher numbers were obtained for the small size class of bacteria with the EUB338 probe when 15% formamide was used than when other levels of formamide were used, and for the medium size class significantly higher numbers were obtained in the presence of 5, 10, 15, and 20% formamide than in the presence of 0 and 30% formamide. When the NONEUB338 probe was used, the numbers obtained in the presence of 15% formamide were significantly higher than the numbers obtained in the presence of 0 and 10% formamide for the small size class and significantly higher than the numbers obtained in the absence of formamide for the medium size class.
The optimal stringency wash time was determined at a fixed formamide concentration of 15% and a hybridization temperature of 37°C. Figure2 shows that the highest counts generally were obtained after 10 min of washing. Hence, for the small size class of bacteria, significantly higher numbers were obtained after wash times of 2 and 10 min than after washing for 60 min. For the medium size class, the numbers obtained after 2, 10, and 30 min of washing were significantly higher than the numbers obtained after 5 and 60 min. Finally, for the large size class (and for the total of all size classes), the numbers obtained after 10 min of washing were significantly higher than the numbers obtained after all other wash times. The lower values obtained after 2 and 5 min were probably due to masking of the bacteria by excess stain when the wash time was too short. Hence, the final standard protocol used for in situ slurry hybridization of indigenous active, soil bacteria included a formamide concentration of 15% in the buffer and a reaction time of 18 h at 37°C, followed by 10 min of washing.
Finally, probe penetration into gram-positive soil bacteria may be difficult and was investigated for Streptomyces sp. andFrankia sp. by using lysozyme treatment (21, 22). To examine this, we dehydrated soil samples with ethanol and then incubated them in a 1-, 2-, or 3-mg ml−1 lysozyme solution. After this, the dehydration procedure was repeated, and the preparations were washed with 1% toluene in 96% ethanol. The lysozyme treatment had no effect on the numbers of cells targeted by the EUB338 probe (data not shown). This is in agreement with results reported by Roller et al. (39) and Zarda et al. (49), who found that lysozyme did not increase the cell counts of gram-negative bacteria. Treatment with lysozyme was therefore omitted in the final standard protocol for in situ slurry hybridization.
Correction model for nonspecific binding of probe.We propose a model for calculating the number of active bacteria in soil (see below) based on the assumption that sufficient unlabelled probe (DNA) should outcompete labelled DNA at both specific and nonspecific DNA binding sites. Figure 3 shows that the cell counts for all three size classes obtained with the EUB338 probe decreased significantly when different amounts of the corresponding unlabelled probe were added. However, no effect of titration with unlabelled probe was observed for the smaller bacteria when they were hybridized with the NONEUB338 probe, indicating that all nonspecific staining in the small cells was due to binding of the rhodamine component of the probe. For medium and large cells, however, the counts obtained with the NONEUB338 probe also decreased as the concentration of unlabelled probe increased. This shows that for larger cells, nonspecific staining caused by DNA binding becomes more important than rhodamine. This is probably due to the lower surface-to-volume ratio of the larger bacteria if it is assumed that rhodamine binding is associated with the cell wall.
The amount of unlabelled probe EUB338 which was sufficient to titrate out binding by DNA of labelled probe EUB338 was defined on basis of the bacteria in the smallest size class because these bacteria were numerically dominant. A constant low number of counts was reached at 12 ng μl−1 (statistically significant), and maximal titration was observed with 36 ng of unlabelled probe per μl for the small size class (Fig. 3). For the medium and large size classes the counts were still decreasing and significantly lower in the presence of 36 ng of unlabelled probe μl−1 than in the presence of 0, 12, and 24 ng of unlabelled probe μl−1; however, at this concentration the counts were only 13 and 9%, respectively, of the counts obtained without unlabelled probe. The numbers of cells in the medium and large size classes represented 7 and 6%, respectively, of the total counts in samples hybridized with labelled EUB338 probe (data not shown). Addition of 36 ng of unlabelled probe μl−1, therefore, reduced the counts to less than 1% of the total counts for the medium and large size classes, and 36 ng of unlabelled probe μl−1 was found to be sufficient to titrate out binding due to the DNA part of the probe.
The model in Table 1 suggests that the numbers of active bacteria determined by in situ slurry hybridization in soil samples can be calculated based on the following four components: (i) homologous binding of the DNA component of the probe to target rRNA; (ii) heterologous binding of the DNA component of the probe to target rRNA; (iii) binding of the DNA component of the probe to nontarget rRNA; and (iv) binding of the rhodamine component of the probe to cell surfaces. In our protocol, the total number of cells determined with labelled, specific probe EUB338 (Nsp) and labelled, nonspecific probe NONEUB338 (Nnon) includes the numbers due to both DNA and rhodamine binding. However, similar samples hybridized with labelled probe plus an excess amount of the corresponding unlabelled probe provide counts (Nsp,unl and Nnon,unl, respectively) resulting from rhodamine binding alone, because the unlabelled probe outcompetes the targets of the labelled probe. To determine the number of cells related to DNA binding alone, the counts related to rhodamine binding are subtracted from the total counts to obtain Nsp − Nsp,unl and Nnon − Nnon,unl, respectively. Finally, the corrected number of active bacteria related to homologous hybridization of the probe to the target rRNA is obtained from (Nsp− Nsp,unl) − (Nnon − Nnon,unl), as indicated in Table 1. While Nspand Nnon depend on the ratio of rhodamine to DNA in both probe EUB338 and probe NONEUB338, the calculated number of active cells depends only on the labelling of EUB338, because components from nonspecific DNA and rhodamine binding cancel each other out.
Detection limit.To determine the number of cells targeted by fluorescence-labelled oligonucleotide probes directed against rRNA, the minimum growth rate which can be determined by fluorescence microscopy has to be considered. We investigated the relationship between growth rate and in situ hybridization signal in soil samples which were amended with formalin-killed bacteria obtained from pure cultures of Arthrobacter globiformis DSM 20124T, Pseudomonas putida MM 3 (16),Pseudomonas fluorescens DF 17 (41), andCorynebacterium sp. strain H 19 (13). Bacteria could be detected easily by the EUB338 probe when the generation times were 5 h or less before the bacteria were added to the soil. In contrast, slowly growing bacteria (generation times, 1 to 2 days or more) could not be detected by the probe (data not shown). The range of generation times between 5 h and 1 day was not investigated.
Bacterial cells were counted with the conventional fluorescence microscope based on staining intensity, cell shape, and color. To test our identification of the active bacteria, selected soil samples were investigated by CLSM by using the improved spatial resolution and narrow emission spectrum of this instrument to give high-quality images. Active bacteria, nonactive bacteria, or nonbacterial soil particles were first identified and assigned to size classes by eye by using conventional fluorescence microscopy. Without moving the microscopic field, we then recorded the images by CLSM. As Fig.4 shows, the particles identified as active bacterial cells by eye had sharper boundaries when they were analyzed by CLSM than the nonbacterial soil particles had, whose shapes were blurred. Furthermore, the light intensity was greater for the active bacteria than for the nonactive bacteria.
Case study of pretreated and untreated soils.The final standard hybridization protocol, including the correction for nonspecific binding, was tested with soil samples representing the following conditions: (i) soil which had been air dried for 2 days and then amended with a glucose solution (final concentration, 3 mg of glucose per g [dry weight] of soil) by rewetting the preparation to field capacity 2 days before hybridization; (ii) soil which had been air dried for 2 days and then rewetted to field capacity 2 days before hybridization; and (iii) soil which had been fixed directly after collection from the field and had a water content of 9.8% on a dry weight basis. The experiment with glucose-treated soil was repeated four times, and the other two experiments were repeated twice. In all repetitions, hybridizations were performed with at least four replicates. The results given below are the means for all experiments for each soil treatment.
Figure 5 shows that the numbers of specifically stained cells (Nsp) decreased in the following order: glucose-amended soil, air-dried and rewetted soil, untreated soil. After the addition of unlabelled probe, the counts (Nsp,unl) were reduced by 35 and 38% in the glucose-treated and dried and rewetted soils, respectively, but only by 6% in the untreated soil. As Fig. 5 also shows, the counts obtained with the nonspecific NONEUB338 probe rarely decreased when excess unlabelled NONEUB338 probe was added; therefore, the nonspecific counts were interpreted as being mostly due to rhodamine binding. In most cases, the nonspecific staining was thus completely compensated for by adding unlabelled probe, so that the numbers of active cells in these soils could be calculated from the counts obtained with the EUB338 probe after subtraction of the counts obtained with the EUB338 probe plus unlabelled probe.
We anticipated that the data obtained for the untreated soil collected under relatively dry conditions would represent the natural low-activity soil conditions under which many small bacteria, but relatively few active bacteria, would be detectable by the in situ slurry hybridization protocol. The results indicate that no active bacteria could be detected in the dry control soil (Fig. 5C), reflecting the detection limit of the method. However, rewetting air-dried soil clearly resulted in higher numbers of active bacteria (3.8 × 108 cells g of soil−1), with the greatest increase occurring in the abundant small-size group of bacteria. Finally, glucose amendment also resulted in a significant increase in the number of active bacteria (4.8 × 108cells g of soil−1), and the small size group was again the predominant group.
The present study shows that the majority of the active soil bacteria, as determined by FISH, are the smallest bacteria (i.e., bacteria with diameters of less than 0.5 μm). The level of activity of these bacteria has not been investigated in detail (45), but the results correspond well with the finding that radiolabelled thymidine and leucine were actively incorporated into such bacteria extracted from soil (4, 5). Addition of a labile carbon source, such as glucose, to soil or rewetting of air-dried soil, resulting in a release of nutrients (38), are both well-known treatments which increase bacterial activity dramatically (8, 9, 13, 47). The increased counts determined after rewetting alone or rewetting plus amendment with glucose are considered evidence that the hybridization protocol and the model for calculating the numbers of active soil bacteria performed correctly.
The rRNA content has been correlated directly with the growth rate for fast-growing bacteria (19, 29), but this relationship may not be valid during slow growth or starvation (12, 30, 44). The relationship between growth rate and sensitivity of the FISH protocol was investigated for laboratory-grown bacteria added to soil. The results demonstrated that cells with generation times of 5 h of less could be easily detected, and the data thus supported the conclusion that the in situ slurry hybridization technique detects only the most active bacteria in soil samples. When flow cytometry was used to quantify light emission by a fluorescein-labelled EUB338 probe, a lower limit of detection was set at generation times of 4 to 7 h for cultures of Escherichia coli and Burkholderia cepacia (46). However, we cannot eliminate the possibility that starved and dormant bacteria may also be detected as active bacteria when their rRNA levels are still relatively high. This has been observed for Salmonella typhimurium(44), and a high level of protein turnover probably accompanied by elevated rRNA levels has been found in starved marine bacteria (31).
In soil samples nonspecific staining may result from binding of either the oligonucleotide (DNA) component or the fluorochrome component of the probe (1, 46). When the FISH technique is used with environmental samples, nonspecifically stained cells are therefore often observed and must be corrected for by subtracting counts obtained by hybridization with a nonspecific probe (28, 33). With soil samples hybridized with labelled probe EUB338 it was not possible to correct for this by directly subtracting counts obtained with the NONEUB338 probe because higher counts were sometimes obtained with the NONEUB338 probe than with the EUB338 probe (Fig. 5). This finding was related to slight differences between the ratios of rhodamine to DNA components in different batches of the probes. To solve this problem, we added different amounts of unlabelled (rhodamine-free) probes to the soil samples in order to titrate out the nonspecific binding of the labelled probe. The contribution of nonspecific binding of DNA was compensated for by subtracting counts obtained with the nonspecific probe from counts obtained with the specific probe after correction for rhodamine binding.
The numbers of active bacteria found in the glucose-treated and the dried and rewetted soil samples represented approximately 5% of the total bacterial counts (5 × 109 to 9 × 109 cells g [dry weight] of soil−1) determined by DAPI staining. This estimate corresponds well with the percentage of active cells determined by the CTC reduction assay performed with bulk soil from the same barley field (48). This is apparently in strong contrast to other reported data, which showed that 41% of the total DAPI counts could be detected with EUB338 probe hybridization by using soil smears (49). However, controls including nonspecific probe NONEUB338 were not included in the previous study, and it is likely that the numbers of bacteria in the samples were overestimated.
The great differences between replicates shows the importance of including at least four replicates with the four combinations of probes in order to obtain consistent results for the different size groups of soil bacteria. The high number of replicates makes the method labor-intensitive and expensive in terms of materials. Additional improvements in the protocol for in situ slurry hybridization should include testing alternative fluorochrome labels for the oligonucleotide probes in order to obtain minimal binding to soil particles and more uniform labelling. To reduce the labor involved, automatic counting procedures based on CLSM, as demonstrated by Bloem et al. (9), could be incorporated into the FISH technique. Despite the obvious limitations at present, including the relatively poor detection limit, our protocol for in situ hybridization to rRNA to determine the number of active soil bacteria is useful and could be improved. The sensitivity for specific groups of bacteria might be increased by performing repeated homogenization-centrifugation extraction for soil bacteria (32). This approach, in combination with soil perturbations, might be used to determine the functions of major soil bacterial groups, such as the Acidobacterium andVerrucomicrobia divisions (25), which have been poorly characterized or not characterized by cultivation methods but are known from 16S rRNA gene sequence comparisons.
Palle Hobolth is acknowledged for synthesizing the oligonucleotide probes. We thank Lars Kongsbak Poulsen and Svend J. Binnerup for their support and advice in developing the in situ hybridization method.
This work was financed by the Danish Center for Microbial Ecology and by the Danish Biotechnology Research Programme 1996-1999 (grant 9502015).
- Received 26 October 1998.
- Accepted 4 January 1999.
- Copyright © 1999 American Society for Microbiology


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State: MI
Zip: 49654
Area: Leelanau
Township: Leland
Acres: 0.75
Topography: Level,Wooded
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|Waste Installed: None
|Waste Options: Municipal
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Listing courtesy of Schaub Team Premier Realty

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Democrats at UN Climate Summit Ignore Science, Reality
A congressional delegation of Democrats at the United Nations COP25 "Climate" in Madrid is ignoring science and the ongoing U.S. withdrawal from the UN Paris Agreement. UN globalists are also exploiting children to advance their phony narrative. In this video, UN expert Debbie Bacigalupi and Wolfgang Muller with the European Institute for Climate and Energy (EIKE) provide a real-time update from the COP25 summit.


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Wait! Who made the Clarion County Photo of the Day?
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"Clarion County Photo of the Day: North Clarion's Cameron Schmader was lucky enough to meet and get a picture with her favorite quarterback, Ben Roethlisberger, on Tuesday at MacBeth's Cabins. The veteran Pittsburgh Steelers QB was visiting Cook Forest with his family." — Clarion County Facebook page.
Aug 21, 2019
August 13, 2019 - RIP Coach Drake
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A prayer before practice for Coach Darryl Drake. Photo: Steelers.
"We're all devastated by (the passing of Coach Drake). I really can't think of any other appropriate words," said Tomlin. "Our time and attention in the recent days has been spent in support of the Drake family. Miss Sheila and Darryl's daughters and extended family – many of us have had an opportunity to spend time with them through this. They've been amazing. In making arrangements and adjusting, we've tried to be as supportive organizationally to them as we can in what is a very difficult time. Professionally the loss is significant, but personally it's even bigger." — Coach Mike Tomlin today.
Aug 13, 2019
August 10, 2019 - A family divided
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If you checked out our Throwback Thursday post on August 8th (link below), you'll see that we have a family divided – Dad is a Browns fan, Mom is a Steelers fan.
But Mom got Dad to agree to take Addison and her little brother Colton to the Steelers Pre-Season game last night….and wait…is that Dad wearing black & gold? Okay, so it's actually a Pirates shirt, but it's a start!!
"Even though it's not a #Browns game, I am as excited as the kids are to be at their first NFL game!" — Ryan Kennedy.
Hmmmmm. We may be rubbing off on him!
Ryan, thank you for being such a GREAT sport!! And Chelsea, thank YOU for persevering!! #HereWeGo
Aug 11, 2019
August 6, 2019 - Random Acts of Kindness
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Katie Stern @ktystern
Lucas, age 4, at @steelers training camp today passing out Terrible Towels to other kids as Random Acts of Kindness in memory of his brother, Toby❤ Can he get a shout out? @TeamJuJu @JamesConner_ @_BigBen7 #thelittlefoxfoundation
Aug 07, 2019
August 6, 2019 - Random Acts of Kindness
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Now when Teresa Varley & Missi Matthews heard that young Lucas was at camp today giving out Terrible Towels to other kids in his brother's memory, they wanted to return the favor and let him meet Ben, James Conner & JuJu.
Teresa shared these wonderful photos with us!
Aug 07, 2019
August 6, 2019 - Random Acts of Kindness
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Now when Teresa Varley & Missi Matthews heard that young Lucas was at camp today giving out Terrible Towels to other kids in his brother's memory, they wanted to return the favor and let him meet Ben, James Conner & JuJu.
Teresa shared these wonderful photos with us!
Aug 07, 2019
August 6, 2019 - Random Acts of Kindness
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"Ben met our son, Lucas, at training camp. Lucas was passing out Terrible Towels as Random Acts of Kindness in honor of his brother, Toby, who died from SIDS in August, 2016. Ben met and talked with Luke on the field after practice, and spoke with my husband about our non-profit, The Little Fox – Toby's Foundation, and our annual Random Acts of Kindness Campaign we do in August in Toby's memory. Thank you to Ben for making Luke's day! It was a moment we will never forget. We know that Toby was smiling down from heaven watching his big brother." www.thelittlefoxfoundation.org — The Stern Family.
Aug 07, 2019
August 2, 2019 - Charlie Batch on social media
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Charlie Batch via Facebook:
This photo was sent to me the other day from practice. Its amazing after 16 years, some things never get old at St. Vincents in Latrobe. Im sure I speak for all of Steelers Nation. Im glad you'll be around for a few more years.
Aug 03, 2019
7/31/19 - "Leading the Way"
119 views
"He's on fire. He's the best, the best there is. I believe when he says that he's going to be the best he's ever been this year, I believe it to be true. — Vance McDonald on #7.
Photo: Karl Roser / Steelers
Aug 01, 2019
2019 - First Weekend of Training Camp
107 views
Mariah with A BIG THANK YOU for Big Ben!
As told by a Steelers fan in attendance:
"This little girl was ecstatic!! I was beside her mom when she came running up and she literally was crying…she was smoooshed from the fans behind her and Ben felt bad for her…one happy little girl!! Ben made alot of fans day by signing for as long as he did…. a day I will cherish forever and ever. Never thought I would get to see him up close!!! OUR STEELERS are a true CLASS ACT!! LOVE THEM ALL!" — Michelle Conley-Bruner
Jul 30, 2019
2019 - First Weekend of Training Camp
113 views
Today was my wife's first day ever at training camp and Ben signed a custom painted football for her. She had a blast and I believe that Jackie will now become a camp regular!! — George Vandergriff.
Jul 29, 2019
2019 - First weekend of Training Camp
104 views
Sonny @TheTerribleBlog
@_BigBen7 and @TeamJuJu definitely made my girl's day by hanging around to do a lot of signing after @steelers camp. Much appreciated!
Jul 29, 2019
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A few years ago, the phrase "nasal sprays" probably made you think of commercials for allergy medications. Yep, we can picture that bee too. But now that more and more nasal sprays made for treating migraine are hitting the market, you're likely starting to get curious about how they actually work.
So we did the research and wrote this guide to answer all of your questions.
First thing's first: How do nasal sprays work in general?
Well, when you spray a treatment into your nose, the medication gets absorbed into the blood vessels in your nostril. After that, it's just like what happens when you take a pill—the medication travels where it needs to go via your bloodstream. So to answer your question, the mechanics of the spray aside, they treat your migraine similar to the way that an oral pill would.
Why would someone choose to spray a medication into their nose when they could just take a pill and get the same effect?
Nasal sprays usually work more quickly than pills because you don't have to wait for them to be dissolved and digested by your stomach, a process that slows during a migraine.
Experts call the time it takes for a medication to kick in "time to peak effect," so nasal sprays have a shorter time to peak effect than tablets do.
For example, if you take a zolmitriptan tablet as soon as you feel a migraine setting in, you could expect to get some relief from your symptoms in about 30 minutes. But if you use Zomig® nasal spray (the same medication, but in nasal spray form), you can get that relief in as little as 15 minutes.
If you've experienced a migraine headache, you know exactly what a big difference 15 minutes can make.
The other factor experts consider when choosing what form to recommend is bioavailability, which means how much of the medication enters your blood circulation after you take it. (That's right—some of what you put into your body never makes it to your bloodstream, which is why vitamins and supplements tend to use higher dosages that you actually need.)
Tablets have the lowest bioavailability, and IV treatments unsurprisingly have the highest. (Fun fact: Orally-dissolving tablets actually take longer to work than standard ones.) Nasal sprays are on the higher end of the spectrum. That means they not only work faster, but are also more efficiently absorbed by your body.
And for people who experience migraine nausea, nasal sprays have an added benefit: You don't have to swallow them ... or worry about vomiting and wasting medication.
So far, three different types of nasal sprays that can treat migraine have hit the market. They're all acute treatments (meaning they relieve symptoms but don't prevent attacks), but they each work a little bit differently.
Like all triptans, nasal triptans relieve migraine symptoms by narrowing overexpanded blood vessels and reducing inflammation in the brain. The nasal triptans that are currently available include Tosymra® (generic sumatriptan) and Zomig® (generic zolmitriptan).
Dihydroergotamine is a formulation of ergotamine, a type of medication that also narrows blood vessels in the brain. (We'll give you a second to digest those two SAT words.) Ergotamine is a natural product of a certain type of fungus. Medically, it's used to treat migraines and cluster headaches. Dihydroergotamine is just ergotamine with hydrogen added to make it more potent.
The only drug of this class that's been approved for migraine treatment so far is Migranal® (generic dihydroergotamine mesylate).
Nasal ketorolac provides a helpful alternative for migraine sufferers with cardiovascular conditions. (It's risky to take medication that constricts blood vessels, like triptans and DHE, if you have any health conditions that affect your blood circulation.) Ketorolac is a type of non-steroidal anti-inflammatory drug (NSAID), a class of pain relievers. Unlike triptans and DHE, nasal ketorolac isn't currently FDA-approved to treat migraine, just pain.
One example of a nasal ketorolac you can buy now is Sprix® (generic ketorolac tromethamine).
Like all acute treatments, nasal sprays work best when you take them as soon as you feel a migraine coming on. While you might be tempted to follow the advice the school nurse gave you when you got a nosebleed, you actually shouldn't tip your head back. Doing so makes the medication drip down your throat and get absorbed by your stomach, which slows down the absorption process. For the same reason, try not to sniff too much while the spray is in your nose.
Since each use should administer a pre-measured amount of medication, you don't need to worry much about dosage. Just be sure not to use the medication more often than you're directed to.
One of the benefits of nasal sprays is that they work quickly enough to be a great option for a back-up medication to use when your first-line treatment just isn't enough. But not all nasal sprays and oral medications can be safely combined.
Regardless of form, different types of triptans can't be mixed, so don't use a triptan nasal spray if you've already taken an oral triptan. In addition, DHE and triptans also shouldn't be used together.
On the other hand, NSAIDs and DHE are not only safe to combine, but are also thought to enhance each other's effect when used together. You can also safely use triptans alongside NSAIDs.
As always, your doctor is the best person to talk to if you have any questions about specific medication combinations.
Like all medications, nasal sprays can come with side effects. Common side effects include:
Talk to your doctor before trying a nasal spray for migraine treatment if you're pregnant or breastfeeding, and if you have high blood pressure, cardiovascular issues, or kidney problems.
Like other recent innovations in migraine treatment, nasal sprays give you a promising new option to consider when you're building your treatment plan. For people whose migraines tend to get very severe very quickly, nasal sprays could make a great first-line treatment. And even if that doesn't describe you, it might not hurt to have a spray as a back-up for your preferred treatment.
The information provided in this article is not a substitute for professional medical advice, diagnosis, or treatment. You should not rely upon the content provided in this article for specific medical advice. If you have any questions or concerns, please talk to your doctor.
Zolmitriptan is an oral medication indicated for the acute treatment of migraine with or without aura in adults and is not used to prevent migraines. Tell your doctor if you are pregnant or plan on getting pregnant. You will need to talk about the benefits and risks of using this drug while you are pregnant. Tell your doctor if you are breast-feeding. Call your doctor right away if you have allergic reactions like skin rash, itching or hives, swelling of the face, lips, or tongue, chest pain or chest tightness, signs and symptoms of a dangerous change in heartbeat or heart rhythm like chest pain; dizziness; fast, irregular heartbeat; palpitations; feeling faint or lightheaded; falls; breathing problems. Call your doctor right away if you have signs and symptoms of a stroke like changes in vision; confusion; trouble speaking or understanding; severe headaches; sudden numbness or weakness of the face, arm or leg; trouble walking; dizziness; loss of balance or coordination. Call your doctor right away if you have signs and symptoms of serotonin syndrome like irritable; confusion; diarrhea; fast or irregular heartbeat; muscle twitching; stiff muscles; trouble walking; sweating; high fever; seizures; chills; vomiting. You can read more about zolmitriptan's side effects, warnings, and precautions here. Full prescribing information for zolmitriptan is available here. You are encouraged to report negative side effects of prescription drugs to the FDA. Visit MedWatch: https://www.fda.gov/Safety/MedWatch/default.htm or call 1-800-FDA-1088.
Sumatriptan is an oral medication indicated for the acute treatment of migraine with or without aura in adults and not for the prophylactic therapy of migraine attacks or for the treatment of cluster headache. Tell your doctor if you are pregnant or plan on getting pregnant. You will need to talk about the benefits and risks of using sumatriptan while you are pregnant. Tell your doctor if you are breast-feeding. Call your doctor right away if you have chest, throat, neck, or jaw tightness, pain, pressure, or heaviness; break out in a cold sweat; shortness of breath; a fast heartbeat; a heartbeat that does not feel normal; or very bad dizziness or passing out. Very bad and sometimes deadly brain blood vessel problems like stroke have rarely happened with this drug. Call your doctor right away if you have weakness on 1 side of the body, trouble speaking or thinking, change in balance, drooping on 1 side of the face, or change in eyesight. You can read more about sumatriptan's side effects, warnings, and precautions here. Full prescribing information for sumatriptan is available here. You are encouraged to report negative side effects of prescription drugs to the FDA. Visit MedWatch: https://www.fda.gov/Safety/MedWatch/default.htm or call 1-800-FDA-1088.
Photo by Windows on Unsplash


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Welcome to the East Launceston Croquet Club
Play for a lifetime and make friends for life
East Launceston Croquet Club
is on Windmill Hill and has been at that site since 1916. This is a designated area for low impact sports. We share the hill with a bowls club and the new aquatic centre. The position is within walking distance of the CBD and there are a number of very good B & Bs in the immediate vicinity.
- The club has regular times for play when you will always find a group to play with
- Both Golf Croquet and Association Croquet are played regularly
- Club events are interspersed between the state and northern events from September to June
- Northern and statewide competitions Competitions are played at the Northern Tasmanian Croquet Centre at St Leonards
- The Club has regular social events throughout the year
Membership numbers are limited as the club has only one court, but membership enquiries are welcome. Annual membership fee is $180, which includes affiliation to Croquet Tasmania and the NTasCC.
Regular coaching sessions are held and individual coaching is available by arrangement. New players will be mentored by more experienced members.
Club colours are green and white or beige. Flat heeled shoes must be worn at all times.
Playing times
Regular social play is currently scheduled for Tuesday, Wednesday and Saturday afternoons and Sunday morning. Members may use the lawn at any other time.
Visitors are welcome to any of these sessions.
Address: Cnr High and Adelaide Streets, LAUNCESTON TAS 7250
PO Box 50, Newstead 7250
President: Alison Henty
Secretary: Sue Walker Phone: 0417 312 718
mail: firstname.lastname@example.com


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AntiquesSanta Fe offers a visual feast for art lovers of all sorts, whether you're a serious collector or a fan who finds joy in just viewing the work.
With more than 250 galleries to explore, you'll be astonished by the city's array of art, including traditional and contemporary painting, photography, pottery, sculpture, textiles and more.
Get a chance to meet some of the artists at Santa Fe's famous Friday evening gallery openings, which take place year-round but with more frequency during the summer months.
As you walk the city discovering one gallery after another along the legendary Canyon Road, around the Plaza and downtown, and in the developing historic Railyard District, you'll agree: art lies at the heart of Santa Fe.
Antiques and Interiors on Grant
Antiques and Interiors on Grant
136 Grant Avenue
Santa Fe, NM 87501
Santa Fe, NM 87501
Multiple Dealer antique and modern furniture, art and decorative objects.
343 Manhattan Avenue
Santa Fe, NM 87501
Santa Fe, NM 87501
Antique home furnishings, folk art, ceramics and devotional art from all of the Americas and beyond.


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A home is probably the largest and most important investment your family will ever make. Protecting that investment is important, and it is an inexpensive way to learn the condition of your home. If you are planning to buy or sell a home, we recommend hiring a professional and certified home inspector to conduct the inspection.
Thank you for your choosing Colorado Elite Home Inspections!
1158 30 1/4 Lane,
Pueblo, CO 81006


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New Manchester United boss David Moyes will turn is attention towards Tottenham Hotspur's flying Welshman areth Bale if his bid to lure Cristiano Ronaldo back to Old Trafford fails.
The former Everton boss and United's top brass will spend the next week testing the waters with Real Madrid to see if they can bring put together a deal in the region of £60million and worth £250,000 a week to the Portuguese attacker.
Club sponsors Chevrolet and Nike are reported to be involved in the package but any move would be contingent on Madrid being satisfied as it is not thought Ronaldo will damage his reputation with Los Blancos supporters by handing in a formal transfer request and newly appointed manager Carlo Ancelotti is not interested in selling.
On taking over at the nine time European champions yesterday the Italian spoke of his excitement at the prospect of working with Ronaldo.
"We've not spoken of signings because today is the first day I am here. I cannot give any information on the players that might come in or those who will leave. The squad is very good. In the next few days we will talk with the club about what we need," he said before making time to single out the club's talisman for praise.
"My opinion is easy. Ronaldo's a fantastic, marvellous player. It's an honour to coach a player like Ronaldo like it was to coach Zidane.
"Ronaldo was not on the list of players like Brazilian Ronaldo, Zidane and Ronaldinho that I coached.
If United fail to turn the player who won three leagues and the Champions League under Sir Alex Ferguson they will tempt Spurs with an £80million bid for Bale, who has yet to sign an improved contract with the club who are desperate to keep him as they attempt to fight their way into the top four.
Spurs chairman Daniel Levy is hope to make Bale the club's top earner and enter into a gentlemen's agreement that he can leave next year but Bale's representative is still sounding out potential suitors to see if they will put up the cash to take him this summer.
Levy employed a similar tactic with Luka Modric two summers ago when Chelsea offered to meet eh club's valuation of the player but the Welshman's 19 goal season has made him many times more valuable than the Croatian.


text: Dave went out of his way to help me make arrangements with a difficult ebay seller that does not understand the meaning of communication. Dave called me regularly to keep me updated on progress (or lack there of), as well as condition of item when he finally picked it up, and was right on with his detailed descriptions. Dave is friendly, patient and willing to work with you the best he can to accomodate your needs. Not to mention he gave me the most reasonable bid for delivering my item. I am so fortunate that I was able to use DMJ Furniture to ship my item from an ebay seller to my doorstep that I would certainly use his services again should the need arise, not to mention I will definately recommend him and his company to anyone needed his services for anything in the future. What a wonderful experience for my first time using uShip! Thank you again, Dave, for taking care of me and my daughter's graduation gift! You Rock!


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Items where Person is "Fernandez-C"
Group by: Date | Item Type | First Author | No Grouping
Jump to: 2015
Number of items: 1.
Gibson, William T. and Gonzalez, Carlos R. and Fernandez, Conchi et al. (2015) Behavioral Responses to a Repetitive Visual Threat Stimulus Express a Persistent State of Defensive Arousal in Drosophila. Current Biology, 25 (11). pp. 1401-1415. ISSN 0960-9822. PMCID PMC4452410. https://resolver.caltech.edu/CaltechAUTHORS:20150514-164948010


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Suboxone Abuse
Suboxone was heralded as being a safe way to treat heroin addition when the prescription drug was introduced in 2003. But after changes in federal laws approved the drug for outpatient use, police in Milwaukee noticed instances of suboxone abuse soon after. Suboxone appeared on the street.
How widespread is the problem of Suboxone Abuse?
A medical director at the pharmaceutical company which manufactures Suboxone reported 15 deaths, in 2009, had been attributed to suboxone abuse. In comparison to the billions of people in the U.S., this is a very small percentage of deaths. In each of the 15 deaths reported nationwide, abusers had combined Suboxone with alcohol or other drugs.
Why would anyone participate in suboxone abuse?
For recreational users, Suboxone combined with alcohol, can produce euphoric effects. Suboxone, by itself, produces euphoric effects. But, the results are short-lived. This is why other sedatives and alcohol are combined with Suboxone to produce deeper, longer periods of euphoria. With combined with other drugs, Suboxone can increase the risk of suppressing breathing and slowing heart rate. This is why the potential to suffer an overdose and the notion of suboxone abuse is a real threat.
What are law enforcement officials doing to curb suboxone abuse?
Undercover detectives have been trying to discourage suboxone abuse by arresting would-be dealers through sting operations. Law enforcement officials aren't surprised to see suboxone abuse in the streets. It is a drug and any drug can be abused. As recreational users find new uses for new prescription drugs approved for outpatient use, officials are hoping suboxone abuse will cease to be a problem.
Do the benefits of using Suboxone outweigh the possibility of suboxone abuse?
In 2009, more than 1,500 patients in Wisconsin were treated using suboxone. These drug treatment programs aimed at stopping opioid abuse usually take from three to six months to complete. Then patients stop using Suboxone and continue their recovery drug-free. Counselors in the Wisconsin area reported success rates ranging from 30 percent to more than 75 percent. Counselors insist there is no opportunity for suboxone abuse using Suboxone alone.
What are the signs of suboxone abuse?
Suboxone abuse is when someone ingests the drug, Suboxone, in combination with other drugs. In combination with alcohol or other drugs, Suboxone can slow breathing and lower heart rates. If you suspect someone of deliberate suboxone abuse, take the victim to a hospital. The threat of an overdose is possible.


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Healing CodesFor The Biological Apocalypse2 DVD SETByDr. Leonard G. Horowitz
Join Dr. Leonard Horowitz as he explores more than 2000 years of religious and political persecution by a secret cabal that has waged ceaseless wars, famines, plagues and propaganda campaigns against Earth's people.
This discussion includes the details about ancient musical and mathematical codes linked to creation, destruction, and miracles, including the repair frequency of DNA: the genetic blueprint of life.
Learn why fungi and prion crystals, associated with Creutzfieldt-Jakob disease, are most likely being spread through infected grains. Like silicon biochips they theoretically provide an electromagnetic frequency receiver and internal energy transmitter to enslave, sicken, and control world populations.
Ancient knowledge and modern science collide in this amazing two DVD set.
UFK476 2 DVD Disks 4 hours $29.95


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by Jarrett Stepman, The Daily Signal
Representative Alexandria Ocasio-Cortez is ready to tax the rich to make her Green New Deal a reality.
"People are going to have to start paying their fair share in taxes," the recently elected New York Democrat told TV show 60 Minutes in an interview.
Speaking of prior decades' taxation rates in the country, Ocasio-Cortez added, "Once you get to the tippy tops, on your 10 millionth dollar, sometimes you see tax rates as high as 60 or 70 percent."
It shouldn't be a surprise that the avowed "democratic socialist" went with the predictable "tax the rich" formula in order to pay for a massive government program to combat climate change.
© 2019 The Heritage Foundation


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The Hussy (1979) Jacques Doillon
Labels : 70's , Drama , France , Jacques Doillon , 0 com
The Hussy (1979)
aka La drôlesse
aka La drôlesse
Genre: Drama
Country: France | Director: Jacques Doillon
Language: French | Subtitles: English (.srt file)
Aspect ratio: Widescreen 1.66:1 | Length: 84mn
Dvdrip Xvid Avi - 688x432 - 25fps - 1004mb
Twenty-something François collects boxes and bottles and resells them to shopkeepers. He lives with his insensitive mother and stepfather. Mado a 11-year old, outsider of her family because of the oddness in expressing her feelings gets kidnapped by François. He takes her to live with him in the attic of his parents' home. Despite the circumstances, an increasingly comforting relationship evolves till the police discovers them. They see the situation under a completely different point of view. Based on a true story.
The Hussy (1979)
If you enjoy this blog, you can support it by buying a NITROFLARE premium account from the links above. For just a few bucks, you will get full speed and unlimited downloads from a great filehost. If you find a dead link, just leave a com on the post REUPS AND REQUESTS and I'll try to fix it as soon as possible. Cheers.


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If you are part of a group that has accommodating members, you're very fortunate. Accommodating describes those who are easy to get along with and willing to help you.
Accommodating is the adjective form of the verb accommodate, and it's used to describe those who are cheerfully willing to make small adjustments to help you out. The waiter who is happy to serve the dressing on the side, give you extra cheese on the burger, and swap French fries for mashed potatoes, all because that's how you want it? Definitely accommodating.


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Class: A4 – LT6B
Number: 24
Essay: Importance of financial statements to managers, investors and creditors
Financial statements are important reports. They show how a business is doing and are very useful internally for a company's stockholders and to its board of directors, its managers and some employees, including labor unions. Externally, they are important to prospective investors, to government agencies responsible for taxing and regulating, to lenders such as banks and credit rating agencies, and to investment analysts and stockbrokers. They are especially useful to managers, investors and creditors.
Firstly, managers of businesses, more than any of the other users, benefit most from the use of …show more content…
Financial information is relevant if it is appropriate to its expected use. It is relevant if used to assist with decisions, including ability to provide feedback and to predict. It its important that the financial statements do not become cluttered with trivial disclosures. Otherwise the overall usefulness of the financial statements will decline.
Thirdly, a business' creditors uses its financial statements in various ways to review and assess the business. By ensuring that its financial statements are accurate, a business can provide useful information to help expedite credit decisions. Each financial statement provides a creditor with a different set of data with which to make a decision about extending credit or to evaluate the credit-worthiness of an existing customer. The balance sheet is a financial statement that provides information about the assets, liabilities and equity of a business. Creditors can use the balance sheet to glean information about the state of a company's cash position, receivables management, inventory management and its debt levels. The balance sheet is a snapshot of the business at a certain place in time. The income statement, also known as a profit and loss statement, provides creditors with vital profitability information about a company. The income statement will provide the


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Try these steps
Open Outlook.
Navigate to Tools - Options - Security Tab.
Click on the Change Automatic Download Settings.
From there choose the options you want i.e. Only from senders on the Safe Senders list or what you like.
Keep in mind that this is a Security option and there can be html attacks received this way.


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Takes One to Know OneLarge Print - 2020 | Center Point Large Print edition
From the critics
Community Activity
CommentAdd a Comment
Corie Geller has what she wanted: she's married to a handsome, considerate and rich man and is mother to his daughter, Eliza. She gave up full time FBI work (as her husband gave up a fancy law practice to become a federal judge) and works in an Arabic language literature niche, doing occasional FBI contract work on the side. Now self-employed, she meets with a group of other self-employed folks on Wednesdays for lunch. She's happy and yet she is vaguely dissatisfied.
Then she begins to notice Pete, who is in the Wednesday lunch bunch. He picks the same seat religiously, watches his car out the window while they lunch, seems incredibly low-key and boring, and yet there is something. He reminds her of her, with her careful bio with attendant details that gives no clue about the FBI. So, using her many investigative skills, she starts asking questions, looking things up, involving her retired police detective father, who does need something to do. Then things begin to heat up.
Just remember the title: "Takes One to Know One."
Disclaimer - I did not finish 'Takes One to Know One'.
The first of the 11 books written by Susan Isaac that I read was 'Shining Through' well over over 25 years ago. I enjoyed it immensely and it has been the touchstone to make me sit up and take notice when I see she's written a new book. 'Takes One to Know One' has diminished that attitude.
My first take away is that the main character is a sad, lonely, confused woman convinced she has a great marriage/life but is constantly whining to herself about how boring her successful, handsome her husband is, how boring her suburban housewife life is, and how she seethes with resentment over living with a phony smile on her face with the choices of hubby's dead wife.
Something about a fellow in her lunch group has tweaked her former life radar and she sets out to find why. It's implausible but the excuse of her boring husband and life are reason enough for her.
The journey through her investigation is mind numbingly boring interrupted by whining and the pretense that she loves her newly privileged life which she relentlessly mocks to herself.
Although I am mildly interested to find out if there is a great mystery and what it might be, I am not interested enough to get beyond the first half of the book. I checked out on page 163. If the story gets better, the author waited too long to get there. I think I gave it a good chance.
A very generous 2 stars
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Ramada Hotel
Ramada Plaza CheongJu is a Premium First Class Hotel that is a 21-story building with 3 lower ground floors and 328 guest rooms, the biggest on in the middle region of korea. It also has a big banquet hall capable of accommodating up to 2 thousands people, Cafe Ramada and other seven restaurants and bars, and a fitness center equipped with a brand-new European outfit. In addition, the restaurants, shopping mall and movie theaters are located within the Hotel complex, which allows our customers to be served as on-stop basis. Ramada Plaza CheongJu Hotel will provide the coziest repose for our customers with our best unique service of warm and sincerity as well as lead your business to a success.
For more information, please click here to get more information about Ramada hotel.
Ree Ho Tourist Hotel
Address: Chungcheongbuk-do Cheongju-si Biha-dong 35-7
- Korea Travel Phone +82-43-1330 (Korean, English, Japanese, Chinese)
- For more info +82-43-233-8800 (Korean)


text:
MainView AutoOPERATOR Basic Automation Guide Volume 1 Using Rules
Contains information about performing system automation with MainView AutoOPERATOR Rules using the Rules Processor application.
- Overview of MainView AutoOPERATOR
- Automation with Rules
- Events
- Using the Rule Processor Detail Control panel
- Creating Rules
- Examples of creating Rules
- Creating more powerful Rules
- Managing Rules and automation using the Automation Control panel
- Using the Rule Set Overview panel to manage Rules in Rule Sets
- Rules Management application
- Field descriptions for the Rule Processor
- Printing Rules and Rule Sets
Was this page helpful? Yes No Submitting... Thank you


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Tag: Basketball
Latest stories
Newcastle Eagles Basketball at Sport Central
A damp Friday morning in Newcastle and I was on route to an even damper Rothbury. Just as I shut the door of my van I spotted a message. "I have 4 corporate tickets for Newcastle Eagles Basketball, for tonight if you fancy them?" We had been racking our brains for something different to do […]


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This is the product description and technical features of the Sievi STAR ROLLER+ S3. This product is found under the Sievi category SAFETY FOOTWEAR.
Technical Information
The Sievi STAR ROLLER+ S3 is a Star Roller is the solution for even the most challenging conditions. To prevent ankle injuries, the ankle part of the shoe is lined with viscoelastic Memory foam padding. The laces are adjusted by turning the dial; the lace mechanism in turn laces the shoes evenly, providing support at the ideal point. New and improved FlexStep® soles maintain their flexibility in sub-zero conditions even better than before, improving the traction of the soles. Metal-free toecaps, thick lining and thermal insoles of the Star Rollers will keep your feet warm. Penetration-resistant steel midsoles provide the best protection against sharp objects..


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It's an industry-wide problem. Flower sales drop in the summer just as flower production is at its peak.
What to do?
A panel of floral industry experts shared tips and resources to address the slump as part of a panel session at Fun 'N Sun, held July 26 in Santa Barbara, California.
The session, "Solving the Summer Slump – A Plan to Drive Sales Beyond Mother's Day," featured Crystal Hedgpeth, floral sales manager for Safeway's Northern California division, John Burk, digital marketing manager for Certified American Grown wholesaler DVFlora and Kelly Shore, owner and lead designer of Petals by the Shore in Maryland. The session was guided by the California Cut Flower Commission's (CCFC) Anna Kalins.
During the session, the panelists explained how they were growing sales and consumer awareness for homegrown flowers by featuring American Grown Flowers and Greens throughout July (declared American Grown Flowers Month by Congress). They also discussed the incredible success they've witnessed with marketing campaigns focused on American Grown flowers and from the American Grown Flowers Month Merchandising Contest – think sales increases as large as 17.5%!
Hedgpeth, Burk and Shore also shared the tactics they and their companies are leveraging to gain customers when "America is in season" and all year long.
Hedgpeth described Nor Cal Safeway's wildly successful blue bucket campaign that promotes American Grown Flowers in dozens of stores. Burk shared DVFlora's multi-pronged efforts, including free marketing materials for its customers, special pricing on American Grown Flowers, DV Crush boxes sent to high-profile florists and special website pages featuring American Grown product. Shore shared her story of committing to sourcing American Grown Flowers for her floral business all year long. She also talked about the many ways she's spreading the word that origin matters, like farm tours, a 12-month project featuring American Grown bridal bouquets and volunteering to lead design efforts for Certified American Grown events.
Attendees left with the knowledge of how to plug into future American Grown Flowers Month campaigns and contests, and a better understanding of why the origin matters message is resonating with consumers.
The big news – tips from the session will help everyone say "so long" to the summer slump!


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Please join us for an interactive convening of Texas philanthropic entities with an interest in children and families. This action-oriented meeting is designed for funders who would like to:
Connect with colleagues and develop a network of funders interested in collaborating to promote innovation in Texas' child welfare system
Learn from leaders in the field about the current environment, best practices, and future opportunities
Take action together to boost the impact of each funder's work
Key content experts include Dr. Peter Pecora, Managing Director of Research Services for Casey Family Programs and Professor of Social Work at the University of Washington and Dr. David Sanders, Executive Vice President of Systems Improvement for Casey Family Programs. The meeting will be facilitated by Dr. Lynda Frost of Lynfro Consulting and will use a repertoire of facilitating methods designed to deeply engage all participants in working together on chronic, entangled challenges.
The first day of the convening will be held at The Meadows Foundation. The second day will be at held Hyatt Place Dallas/North Galleria. An agenda will be available on the PSW website in a few weeks. For program questions, please contact Luanne Southern or Lynda Frost.
Access resources program materials here.
Additional resources: Building Bridges Initiative
Registration is now closed.


text: To begin earning commission on your referrals please complete the form below. As a Referral Partner you can receive a 10% commission on all uCollect accounts that are created with your Referral Code. You may share that with your customer by way of a discount on their subscription (you can choose any allocation, as long as the total does not exceed 10% - this way you can look good by saying tha you have negotiated a discount to your clients). You can also nominate some accounts to be billed to your Partner Account and charged to your Credit Card each month rather than being billed to the organisation directly.To create your Referral Code please complete the form below and one will be emailed to you:


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Symptoms of Alcohol Detox
The symptoms of alcohol detox or alcohol withdrawal may range from minor to severe and can even be deadly if they go untreated. Alcohol detox symptoms are most often uncomfortable and difficult for the addict to deal with which is why most alcoholics continue to drink despite realizing the negative consequences that result from their drinking each day. Alcohol detox symptoms can begin as soon as just a few hours after the individual takes their last drink but are most prevalent during the second and third day following the abrupt quitting of alcohol use.
One may think that alcohol detox symptoms are only present for those who drink heavily every day but this is not the case. Many of the symptoms of alcohol detox affect individuals who have only had a few drinks occasionally and those who have a few each day or those who drink heavily but not everyday. Alcoholism and the symptoms of alcohol detox can affect any individual from any walk of life, rich or poor, young or old, heavy drinker or mild to moderate drinker.
Beginning Symptoms of Alcohol Detox
When an individual who is physically dependent on alcohol abruptly stops drinking they are subject to a range of alcohol withdrawal symptoms that can be mild to moderate or may even become severe. The beginning symptoms of alcohol detox may include the following mild symptoms of alcohol detox:
- nausea
- headaches
- anxiety
- shakes
- sweats
These alcohol detox symptoms are mostly not life threatening but they can be rather uncomfortable for the individual and the headaches, shakes and nausea especially may cause the individual to start drinking again in an effort to stop the uncomfortable symptoms. Some additional symptoms of alcohol detox that could become life threatening include increased heart rate and increased blood pressure. These symptoms can all be treated when an individual is properly monitored for alcohol detox symptoms in an alcohol detox center.
Alcohol Detox Symptoms: The Most Common Symptom that Goes Unnoticed
There is one very common alcohol detox symptom that tends to go unnoticed or untreated and for some reason does not alert the individual to seek help—the hangover. When an addict feels hungover they are not thinking about the fact that a hangover is a very common alcohol detox symptom. Hangovers may include nausea, vomiting, sweats, and shakes, all the symptoms of alcohol detox. Hangover is the number one alcohol detox symptom that does not get recognized as alcohol detox but the morning after hangover is a mild to moderate form of what an individual will feel when they abruptly stop drinking and begin to detox from alcohol.
Serious Alcohol Detox Symptoms
Although it is rather rare that alcohol detox symptoms get so severe that medical intervention is required to assure the safety of the addict it is possible. Alcohol detox symptoms, in rare cases, can become so severe that without medical intervention and special medications the alcohol withdrawal symptoms can result in death. The most severe alcohol detox symptoms include:
- hallucinations
- convulsions
- seizures
- fever
When these alcohol detox symptoms are not properly treated the end result can be very dangerous for the addict. Because of the potential for alcohol detox symptoms to become so severe it is recommended that all addicts who try to stop drinking seek the help and medical care of an inpatient alcohol detox center that can provide around the clock medical supervision and intervention when necessary to assure the utmost safety of the addict.


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Leave us a review!
Club Reviews
Completely different from every club we have ever been to!
We have been to a number of clubs all over the southeast, as well as house parties and hotel takeovers. None have compared to what has been created here. The atmosphere is impressive. The hosts were incredibly friendly, and introduced us to club members, other hosts, and even the owners! We felt extremely welcomed by any and everyone we spoke to! The club was clean, and well kept. They even served dinner there! The themed rooms and areas have so much character and are well thought out. We cant come up with a single complaint about this place and plan to make every party possible. The Oasis is also the most cost effective club we have been to. No need for a hotel after the club, we dont have to grab a meal before going, and the entry is lower than anywhere we have been for a saturday night party! We will attend here as long as its doors are open!!
A place with no judgment, sexy friendly welcoming atmosphere!
We have been to other clubs and not one can take the place of E&C Oasis. By far it is the best club we have been to. The comfort we feel here is like being at home. The owners Ellis and Char are the sweetest friendliest club owners we have ever met. The host and hostesses are crazy and fun as hell to be around. Never a dull party! So many friendly open minded guests with respect. E&C Oasis is the place to be.
Great atmosphere
This club has a great atmosphere. Owners and host are great people. Great place to party.
Great place for ALL to enjoy!
I had a wonderful first time and experience at E&C. The owners & hosts/hostess did an amazing job. The place is so inviting and none judge mental. The rooms are unique and clean. You can definitely let loose and be yourself while indulging in your wildest fantasies.
Friendly guests. Clean play areas.
E&C has become a second home to me. The owners/managers are truly some of the greatest people I know, and I consider them family. The play rooms are always clean, and the care put into each one's theme is a special touch you won't find anywhere else. Every party is different, and I can honestly say the guests have always been friendly, respectful, and welcoming.


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Dental Healthcare Technology
We build and integrate scalable, enterprise-level, cloud-based, web applications and software as a service (SaaS) platforms for the dental industry.
Denti.net integrates with the apps you rely on to manage your practice and operate efficiently
"Thanks for all your hard work on the website. The website looks great."
Dr. Brian Queen, DDS
Blogging Tips for Dentists and Dental Office ManagersPosted by Blake Newman in Online Marketing for dentists, SEO for dentists Tagged in blog ideas for dentists, can a loose tooth be saved, how to overcome bloggers block, how to overcome writers block
How to rank on search for "dentists near me"Posted by Blake Newman in How to, SEO for dentists Tagged in dentists near me, dentists who take medicaid near me
Contact Us
Call for FREE consultation: +1 (202) 660-2940


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Car Seat fitting for Monday to Friday only.
A fitting is the installation of a car seat, relocate position of car seat or refit a car seat to rearward/forward position.
Fitting ranges from approximately 10 minutes to 20 minutes.
Parts and accessories not included.
Most installation of Child Car Seat can be done without additional parts. Some car seats and some older model cars may need additional parts for correct installation.


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I burnt a disc using DVDFab HD Decrypter. I'm using DVDFab version 8.2.1
This disc works well on any DVD player I have including the computer. I cannot view this disc on my Sony Blu-Ray player. I have Windows 7 64bit Professional. As I said, it burns very well but I just can't view the DVD on the Sony Blu-Ray.
When I used version 220.127.116.11, I was able to view the results on the Blu-Ray player. The Blu-Ray is Sony BDR-S350.
It seems that there are problems with ripping DVDs using the new version of the software. From what I understand, the Blu-Ray player does not recognize the ripped disc. I would like to suggest that you should wipe clean the surface of the DVD because the player may encounter problems if it's scratched. Plus, these Blu-Ray players are very sensitive due to the fact that they demand high quality DVDs for their full performance and image quality. On the other hand, another possibility is that the new version of the ripping software did not rip the original disc as it should or the settings were not properly configured. From the settings menu of the software, wipe all the settings by returning to the default ones using the integrated option Restore to default settings.


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Cowboy Bebop Completed 4.66
Author: YATATE Hajime
Action Adventure Comedy Sci-fi
Spike, Jet, Faye and Ed - cowboys on the new frontier. Together this band of interplanetary bounty hunters takes on the jobs that anyone in their right mind would turn down.
CHAPTER(11) Last Updated: Oct 16,2007
FACEBOOK COMMENTS MANGA HERE COMMENTS (0) Newest Oldest Popular


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Recommended planner application - Efficient Calendar
Efficient Calendar is an attractive, award-winning and elegant planner application that can help you plan and manage your time. It is a combination of calendar and task tracking system with project management and To-Do lists.
- Main Holidays Can Be Imported at Any Time
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- Easy to Install
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- Recognition from Authoritative Software Editors
- Calendar and Time Manager
- Recycle Bin
- Edit Feature Similar to Word
Latest Version: 5.60 Build 559
Price: $34.95
Requirements: Windows 10/8/7/XP/Vista, Android, iOS
Languages: English, Indonesian, Arabic, Portuguese, Finnish, Traditional Chinese, German, Polish, Swedish, Croatian, Slovenian, Japanese, Korean, Turkish, Italian, Spanish, Norwegian, Ukrainian, Farsi, Danish, Greek, Hungarian, Thai, Armenian, Dutch, Lithuanian, French, Russian, Bulgarian, Simplified Chinese, Czech
Other planner applications provided by Efficient Software
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Create and manage various events and to-do list in your computer with the attractive free calendar software program. Read More...
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Manage your contact information, time, calendar and other personal information with the flexible personal information manager. Read More...
Efficient Man's Organizer $39.95
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|Spell checker
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|To-Do List & Project & Task Manager
|Sounds Good?
|Efficient Man's/Lady's Organizer
|Efficient Calendar
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"What was the biggest influence on your decision to purchase EfficientPIM? My answer: Free trial worked well for my needs. Please write down your comments on EfficientPIM. My answer: Have found it useful for computer access to my address book and general personal organizational tasks. Love that I can access future calendars far in advance."
- Gail Holmes, August 2012
"Pros: 1. Easy to use. 2. Nicely designed. 3. Includes calendar, task list, contacts list, diary, notes and more."
- Softonic
"I find this program has everything I need. Pros: Easy to use. I use this as a journal to reference past appointments. Excellent! It is always there, I cannot misfile or forget what I saved the file as. Cons: Need smaller time frames for better scheduling. 5, 10, 15. Summary: A very good program. Saves time and keeps track of items that need remembering."
- 54susieq, May 2011


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Intensive glucose control was no help for reducing long-term risk of cardiovascular events, according to 15-year data from the landmark Veterans Affairs Diabetes Trial (VADT).
Among 1,665 military veterans with type 2 diabetes, intensive glucose control was not significantly better for reducing risk of major cardiovascular events compared with standard glucose therapy (HR 0.91, 95% CI 0.78-1.06, P=0.23), reported Peter Reaven, MD, of Phoenix VA Health Care System in Arizona, and colleagues.
This outcome was also independent of baseline cardiovascular disease risk, the group explained in the New England Journal of Medicine.
Similarly, intensive glucose control wasn't any better for reducing the risk for death from any cause during the 15-year follow-up period (HR 1.02, 95% CI 0.88-1.18). Other secondary outcomes, including risk of death from only cardiovascular causes (HR 0.94, 95% CI 0.73-1.20) and any major diabetes outcome (HR 0.90, 95% CI 0.78-1.04) also weren't significant between intensive versus standard therapy.
There also wasn't any significant health-related quality of life benefit to intensive therapy, either. While a five-point difference was needed to be considered clinically meaningful, there was only a 1.6-point difference between the groups.
These findings don't come as much of a surprise, as the initial findings from VADT also found no significant reduction in these events with intensive glucose control over a median 5.6-year intervention period.
In the initial trial, which included 1,791 military veterans with type 2 diabetes, half were randomized to receive intensive therapy and the other half to standard therapy. Those on intensive therapy received slightly higher doses of thiazolidinediones, insulin, oral sulfonylureas, and acarbose, compared with standard therapy.
However, during active treatment, those receiving intensive management saw a significant weight gain marked by an average of a 1.3 point higher BMI.
After the active intervention period, those on intensive therapy had a 1.5% lower HbA1c level than those on standard therapy (6.9% vs 8.4%), although this gap closed within 3 years after the active intervention ended.
Prior to this gap closing, and during the nearly 10-year period where the intensive therapy group had significantly lower HbA1c levels, they also had a 17% reduced risk for primary cardiovascular events (HR 0.83, 95% CI, 0.70-0.99). Only after the intensive therapy group's A1c levels rose did they actually begin to see a slightly higher risk for major cardiovascular events compared with the standard therapy group (HR 1.26, 95% CI 0.90-1.75).
"Because the duration of separation of the glycated hemoglobin curves during our original trial and the post-trial observational follow-up was long, our findings of no legacy effect and no mortality benefit expand on similar results in the ACCORD and ADVANCE trials, which also involved older patients with well-established type 2 diabetes," the researchers explained.
However, one major limitation to the study was the inclusion of mostly older men. "[O]ur results are generally consistent with the findings of the ACCORD and ADVANCE follow-up studies, which had greater representation of women," they noted, adding how it's "possible that underlying atherosclerosis and cardiovascular injury were too advanced in these participants to be effectively altered by glucose lowering."
Reaven's group also noted that because of the post-trial follow-up design of this analysis, they were unable to measure for any potential differences in other long-term diabetes-related outcomes, such as neuropathy or retinopathy.
The study was supported by the Office of Research and Development of the VA Cooperative Studies Program. The primary VADT received support from the VA Cooperative Studies Program, the American Diabetes Association, and the National Eye Institute.
Reaven disclosed relevant relationships with AstraZeneca, Bristol-Myers Squibb, Novo Nordisk, Sanofi, Boston Heart Diagnostics, and Takeda Pharmaceutical. Emanuele disclosed a relevant relationship with Merck.
Primary Source
New England Journal of Medicine
Source Reference: Reaven P, et al "Intensive Glucose Control in Patients with Type 2 Diabetes -- 15-Year Follow-up" N Engl J Med 2019; DOI: 10.1056/NEJMoa1806802.


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Faculty Advisor. Duke Disability Alliance. 2017
Event/Organization Administration
Duke Disability Alliance promotes accessibility and inclusion for students with disabilities and their allies, both on campus and in the community.
Service Performed By
- Quirici, Marion L Lecturing Fellow of Thompson Writing Program
- Faculty Advisor
- 2017
Host Organization
- Duke Disability Alliance


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You may already be familiar with the basics of what to do and what not to do in designing a promotional brochure for advertising and marketing that is effective. Now, you might be wondering what else you can do to make your brochure stand out more. Don't worry, we got your back. In this article, you can find a few other things that you may haven't really thought to do for your brochure printing.
1. Add a call to action
A call to action statement is important, and a lot of people are still not very aware of this aspect. Making a clear call to action direction ensures that your reader will know what to do after they read the information displayed on the brochure in case they are interested. The most common example of a call to action statement is "Call us!" or "Order now!". However, you can always be creative and make your own call to action statements. Speaking about a call to action, you can even engage more with your reader or potential customer by printing a QR code on your brochure that can be scanned by smartphones. Make this QR code direct your potential customers to your website or social media so that they can learn more about your business or promotions beyond what is displayed on your brochure. A brochure like this can leave a long-lasting impression that is also strong.
2. Include positive testimonials or reviews
You can try putting positive feedbacks that you have earned if available to let people know that your product or service is reliable. The testimonials or reviews that you put on your brochure must be ones that are well written and points out your strengths well. It is even better if a person with power or capability of influencing others can leave a testimonial for you. In order to do this, you usually have to partner up with them and endorse an influencer. However, this surely will cost you more thus it isn't mandatory.
3. Choose an appropriate finish
There are several printing finishes that you can choose to have for your brochure. Most commonly, the brochure is given paper coatings at the end of its printing process. This can make a brochure last longer and it enhances the overall look of the brochure itself. A coating usually makes the colors pop more, making your brochure look more vivid and interesting.
4. Look out for bleed and trim area
In brochure printing, you will encounter the step of cutting. In this cutting step, your brochure will be cut to its desired shape. To avoid mistakes from happening in this step, during the designing process, you have to make a spare space for bleed around the border area of the brochure. This bleed area will then get trimmed during the cutting process, leaving only the desired part of your brochure to remain. This ensures that your brochures will come out consistent, and there won't be any important part of getting cut off during the brochure cutting step.


text: If you want to serve dinner in the most inspiring setting, look no further than Lotus Dining Chairs. These upholstered dining chairs are the more comfortable stylish option when it comes to dining. These chairs are waterproof, sturdy and easy to clean as well as available in brown and ruby colours. Each seat is wide, comfortable along with foam cushioning back for total comfort. Their back curves gently for an effortlessly tailored classic style.


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LOOKOUT MOUNTAIN -- Freshman baseball player Scott Gillespie and senior softball player Chelsea Krafft have been named the Covenant Players of the Week for the week of March 12-17.
Gillespie had an outstanding four games last week as Covenant went 2-2. The freshman hit .643 with nine hits in 14 at-bats, four doubles, and eight runs batted in. He also walked three times and struck out just once. In the finale of a three-game series at Huntingdon College on Saturday, Gillespie went 3-for-3 with five RBIs and a run scored. For the season, Gillespie is second on the team in batting average at .382 with six doubles and 16 RBIs and a .471 on-base percentage.
On the women's side, Krafft wins the award for the second straight week. Last week, Krafft pitched complete-games in all four of the Lady Scots' contests going 2-2 with an earned run average of 1.32. Krafft pitched 27.1 innings and allowed just four earned runs on 21 hits, while striking out 25 and walking five.
May 18, 2012 Lady Scots Softball Earns More Accolades
May 7, 2012 Lady Scots Earn NCCAA At-Large Berth
April 30, 2012 Lady Scots Softball Ousted in NCCAA Regional
April 29, 2012 2012 NCCAA Softball Mid-East Region Tournament
April 26, 2012 Lady Scots Softball Receives GSAC Awards
April 25, 2012 Hannah Rogers Heads South
April 25, 2012 Softball Adds Maryland Pitcher
April 25, 2012 No. 2 Emory Cruises Past Lady Scots
April 22, 2012 Lady Scots Fall on Senior Day, 5-3
April 18, 2012 Rollins and Wagoner Named Covenant Players of the Week
April 18, 2012 Lady Scots Split Doubleheader at Maryville
April 12, 2012 Hyatt and Redberg, Krafft Win GSAC Weekly Awards
April 11, 2012 Berry Rallies to Sweep Lady Scots
April 10, 2012 Rollins and Harris Named Covenant Players of the Week
April 3, 2012 Covenant Trumps Tennessee Temple
April 3, 2012 Hirte and Wagoner Named Covenant Players of the Week
April 3, 2012 Covenant Athletic Weekly Recap: Week 28 (March 26-31)
March 31, 2012 Softball Loses Home Twinbill to LaGrange
March 28, 2012 Timely Hitting, Pitching Gives Covenant Sweep of Judson
March 27, 2012 Win-Streak Snapped; Lady Scots Split With Berry
March 24, 2012 Covenant Earns Two More Conference Wins
March 22, 2012 Lady Scots Sweep Spelman
March 21, 2012 Gillespie and Krafft Named Covenant Players of the Week
March 21, 2012 Covenant Athletic Weekly Recap: Week 26 (March 12-17)
March 19, 2012 Krafft and Harris Combine for No-No, Covenant Sweeps
March 17, 2012 Softball Gets First Two GSAC Wins With Sweep
March 13, 2012 Piedmont Sweeps Covenant
March 13, 2012 Covenant Players of the Week Awards go to Krafft and Smith
March 10, 2012 Lady Scots Sweep Previously Unbeaten Birmingham-Southern
March 6, 2012 Hilt and Johnson are Covenant Players of the Week
March 5, 2012 Covenant Athletic Weekly Recap: Week 24 (Feb. 27-Mar. 3)
March 3, 2012 Rain Washes Away Weekend Events
February 28, 2012 Harris and Hirte Earn Covenant Players of the Week
February 28, 2012 Covenant Athletic Weekly Recap: Week 23 (Feb. 20-25)
February 27, 2012 Covenant Softball adds Kendall Whann to 2012-2013 Roster
February 21, 2012 Softball Set to Start New Season
February 9, 2012 Softball Secures a Spot for Madison Quirk
February 6, 2012 Women's Softball Catches Another Early Commitment
November 7, 2011 Softball Adds Morgan Lane to Infield for 2013 Season


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365 days of SL, day 221:
(Click for larger — 1920x1033 PNG, 2029 KiB)
"Dear Princess Celestia, last night I learned that some people are utter twats..."
Alas, it's true. As planned, I went back to the aicraft carrier seen yesterday; I poked around for a bit, and although it was mostly empty, I thought it was an interesting (and extensive) build, anyway. In the end, we found the commander's office, and proceeded to make ourselves at home.
The guy owning the parcel actually showed up after a while. Usually, this is an opportunity to make a bit of smalltalk, but he turned out to be to a five-star asshat, so after having to put up with some entirely unexpected (and undeserved) nastiness, I left, and he was added to my killfile. Plonk!
Unfortunately, by this time, it was already very late, and I didn't feel like using the screenshot I'd already taken earlier anymore; we went to a cave below a friend's parcel for a bit, and I took a quick one there, but I'd be lying if I said it didn't suck. Oh well; c'est la vie, and the one I had earlier wasn't stellar, either.
Fortunately, unpleasant people, by and large, are in the minority in SL, so maybe tonight will be nicer again, and yield a nicer screenshot.
Location: Jaguar Valley (34, 54, 21) (Mass Hysteria)