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Candida parapsilosis is known to show limited genetic variability, despite different karyotypes and phenotypes have been described. To further investigate this aspect, a collection of 62 sensu strictu C. parapsilosis independent isolates from 4 geographic regions (Italy, n = 19; New Zealand, n = 15; Argentina, n = 14; and Hungary, n = 14) and different body sites (superficial and deep seated) were analysed for their genetic and phenotypic traits. Amplification fragment length polymorphism (AFLP) analysis was used to confirm species identification and to evaluate intraspecific genetic variability. Phenotypic characterisation included clinically relevant traits, such as drug susceptibility, in vitro biofilm formation and aspartyl protease secretion.
AFLP genotyping showed little variation among isolates, when the presence/absence of bands was considered. However, when AFLP profiles were compared by relative intensity for each fragment, a significant level of variation and geographical clustering was observed. All isolates were found to be susceptible to commonly used antifungals, although a reduced susceptibility to echinocandins was observed in all isolates. C. parapsilosis isolates from different geographic origins varied in the number of biofilm producers, with a higher prevalence of producers isolated in Hungary and Argentina. The frequency of secreted proteinase producers also varied in isolates obtained from different areas, with a higher number of proteinase producers found in Italy and New Zealand. Interestingly, biofilm production and proteinase secretion were negatively correlated. This finding could be explained by assuming that proteinase activity plays a role in detachment and release from a mature biofilm, via degradation of C. parapsilosis adhesins and/or extracellular matrix components, as observed for other microorganisms.
The low number of polymorphic AFLP bands (18 out of 80) obtained for C. parapsilosis isolates is in agreement with the limited sequence variability described for this species. However, when band intensity was included in the analysis, geographical clustering was observed. Expression of virulence factors varied among strains isolated from different geographical regions, with biofilm and proteinase producers more frequently isolated from Hungary and Italy, respectively.
Of the species belonging to the "psilosis" group, Candida parapsilosis is by far the most studied and characterised. It represents about 90% of the infection attributed to C. parapsilosis sensu lato [1] and it seems to be better adapted to the human host than the two relatives (C. orthopsilosis and C. metapsilosis), as also shown by the high incidence of C. parapsilosis systemic infection worldwide, assessed as the second most common candidemia in many countries [2-6]. C. parapsilosis is an opportunistic pathogen that colonises human skin and can spread nosocomially through hand carriage [7,8]. It has been frequently associated with infections in newborns [6,8,9] and in catheterised patients [3]. This can be linked to the ability of C. parapsilosis to produce biofilm in the presence of plastic surfaces such as catheters or other prosthetic materials [6,10-12].
An increasing number of studies points towards a reduced genetic variability among C. parapsilosis isolates, which has been interpreted as a predominant clonal mode of reproduction [6,13-15]. This is in contrast to what has been recently described for C. metapsilosis and C. orthopsilosis species, in which recombination has been shown to occur by AFLP analysis [16,17]. On the other hand, a notable variability in virulence phenotypes has been observed for C. parapsilosis, such as the ability to produce biofilm or hydrolytic enzymes [6,18].
In this study, a selection of 62 C. parapsilosis isolates obtained from different patients was made from a wide collection of "psilosis" isolates on the basis of different geographical (Italy, Hungary, Argentina, New Zealand) and anatomical (blood, mucosa, nail, cerebrospinal fluid, etc) origins, in order to evaluate if these factors may have an impact on genetic variability.
AFLP was applied to our entire "psilosis" collection (n = 650), as this method has been shown to reproducibly and unequivocally identify Candida species [16,17,19]. The 62 selected isolates were analysed further by using another enzyme/primer combination EcoRI-HindIII, since the previously used EcoRI-MseI combination was found to be less discriminative and affected by band homoplasy in C. parapsilosis and C. metapsilosis [unpublished data, [17]]. The EcoRI/HindIII enzyme combination gives rise to larger fragments and therefore increases the sensitivity to detect polymorphisms.
In parallel, phenotypic properties such as biofilm formation and proteinase secretion were analysed. Since the "psilosis" species have been recently associated with a lower susceptibility to the echinocandin class of antifungals [20,21], drug susceptibility was also evaluated and extended to other antifungals. The overall goal of this study was to gain further information on genotypic and phenotypic properties of this successful and yet elusive opportunistic pathogen.
The Candida parapsilosis collection included 62 individual isolates obtained from different body sites and geographical regions (Table 1). The majority of Italian isolates (n = 19) was provided by the Unità Operativa di Microbiologia, Ospedale Universitario, Pisa; 6 isolates being from different Italian hospitals (Table 1). Hungarian isolates (n = 14) were from the Department of Microbiology, Medical School, Debrecen. Argentinian and New Zealand isolates were kindly provided by Dr Marisa Biasoli, Centro de Referencia de Micologia, University of Rosario and by Dr Arlo Upton, Auckland City Hospital, respectively. The isolates used in this study were initially identified as C. parapsilosis according to their biochemical profile on API32 ID and a Vitek 2 advanced colorimetric semi automated system (bioMérieux, Marcy l'Etoile, France). C. parapsilosis ATCC 22019 was included in the study as reference strain. All isolates were maintained on Sabouraud agar (Liofilchem S.r.l., TE, Italy) for the duration of the study.
Table 1. Details and phenotypic properties of Candida parapsilosis clinical isolates used in this study.
DNA extraction and molecular typing of Candida parapsilosis
Genomic DNA was extracted from yeast samples grown in Sabouraud broth, (Liofilchem) as previously described [16]. DNA quantity and integrity was assessed by gel electrophoresis.
AFLP analysis was used to confirm species identification and to evaluate the genetic relatedness of C. parapsilosis isolates. AFLP was performed on 50 ng of genomic DNA as previously described [16]. The restriction-enzyme combination EcoRI/HindIII was used in the first restriction/ligation step. The concentration of the HindIII adaptor was equal to EcoRI (0.45 μM). Sequences of the adapters and pre-selective primers used for AFLP analysis were as already reported [17]. Pre-selective, selective amplifications and gel electrophoresis conditions were performed as previously described [16]. AFLP profiles, ranging from 100 to 700 bases, were exported as a TIFF file and analyzed with the TotalLab TL120 software package (Nonlinear Dynamics Ltd, UK) to evaluate genetic variability within the species. DNA bands obtained for each isolate were size-matched. AFLP bands were defined by time (Rf value) and by the surface of the fluorescent peak they form, as recently described [17]. Only bands which were at least 0.5% of the lane volume present in at least one of the isolates were included in the analysis. Bands were considered to be absent as the surface of the peak was less than 0.03% of the lane volume.
Dendrograms were built by the TL120 software using the unweighted-pair group method using arithmetic means (UPGMA). For each pair of isolates, a similarity index (SAB) was calculated, ranging between 0 (complete non-identity) and 1.0 (identity). The SAB between the patterns for every pair of isolates A and B was computed by the formula SAB = 2E/(2E+a+b), where E is the number of bands shared by both isolates A and B, a is the number of unique bands in the pattern for isolate A absent in the pattern for isolate B, and b is the number of unique bands for isolate B not present in isolate A.
Since C. parapsilosis isolates displayed very little polymorphic fragments, but showed a great variation in band intensity, the latter parameter was included in genotype analysis. Thus, the quantity of each AFLP fragment was normalised as a percentage of the total quantity of the AFLP fragments for a given isolate and defined as relative intensity. For each isolate pair, the Pearson's correlation of the relative intensities % of all fragments present in the two isolates was determined: a correlation index of 1 corresponded to a complete identical pattern.
A distance matrix was obtained by subtracting the correlation between two AFLP patterns from 1 (distance = 1-correlation). This distance matrix was imported into the Treefit program [22] and used to produce a UPGMA dendrogram, which was visualised with the Treeview program [23,24].
Biofilm formation
Biofilm production by C. parapsilosis clinical isolates was evaluated as previously described [16,17]. One set of plates was incubated at 37°C and another at 30°C without agitation. After 24 h, plates were washed and the optical density was measured (OD at 450 nm). Biofilm production was considered as absent (no production; NP), when the OD at 450 nm was lower than 0.03, weak (WP, 0.03 ≤ OD < 0.08), moderate (MP, 0.08 ≤ OD < 0.16), or high (HP, OD ≥ 0. 16) [16].
Proteinase secretion assay
Yeasts were pre-grown in YEPD liquid medium (2% glucose, 1% yeast extract and 2% bactopepton, Difco, Detroit, MI, USA). C. parapsilosis isolates were analyzed for secreted proteolytic activity on solid medium containing bovine serum albumin (BSA) as the sole nitrogen source. The inducing medium containing 1.17% yeast carbon base (Difco); 0.01% yeast extract (Biolife, Milan, Italy); 0.2% BSA (pH 5.0) (BDH, Poole, UK) was sterilised by filtration and added to a solution of autoclaved (2%) agar. The number of blastoconidia was microscopically determined and yeast suspensions were adjusted to 106cells/ml. Ten μl of each yeast suspension was inoculated in duplicate onto BSA agar plates and incubated at 30°C for 7 days. Proteolysis was determined by amido black staining of the BSA present in the medium as described by Ruchel and colleagues [25]. Proteinase activity was considered to be absent when no clarification of the medium around the colony was visible (radius of proteolysis < 1 mm), weak when a clear zone was visible (1 ≤ radius < 2 mm), moderate when the clarification radius was comprised between 2 and 3 mm and high, when the proteolytic halo exceeded 3 mm in radius.
Antifungal susceptibility
The colorimetric broth micro dilution method SensititreYeastOne® (YO-9, Trek Diagnostic Systems Inc., Cleveland, USA) was used to evaluate C. parapsilosis susceptibility to amphotericin B, fluconazole, posaconazole, itraconazole, voriconazole, 5-flucytosine and the echinocandins (caspofungin, micafungin, anidulafungin) as previously described [17]. According to manufacture instructions, the positive growth well was examined after 24 hour incubation. If the well was red, endpoint for antifungal could be interpreted, otherwise plates were incubated for a further 24 hours.
Antifungal susceptibility interpretation criteria were according to the Clinical Laboratory Standards Institute (CLSI) M27-A3 and M27-S3 documents [26,27]. Briefly, caspofungin MIC ≤ 2 (μg/ml) susceptible (S) and > 2 (μg/ml) non susceptible; fluconazole MIC ≤ 8 (μg/ml) S, MIC between 16 and 32 (μg/ml) susceptible dose dependent (S-DD), MIC ≥ 64 (μg/ml) resistant (R); itraconazole MIC ≤ 0.125 (μg/ml) S, MIC between 0.25 and 0.5 (μg/ml) S-DD, MIC ≥ 1 (μg/ml) R; voriconazole MIC ≤ 1 (μg/ml) S, MIC = 2 (μg/ml) S-DD, MIC ≥ 4 (μg/ml) R; amphotericin B MIC ≤ 1 (μg/ml) S; 5-flucytosine MIC ≤ 4 (μg/ml) S, MIC between 8 and 16 (μg/ml) intermediate (I), MIC ≥ 32 (μg/ml) R [25,26]. For S-DD category, susceptibility is dependent on achieving the maximal possible blood level of the antifungal [26,27]. C. parapsilosis reference strain ATCC 22019 was used as control.
Statistical analysis was performed using Instat software (GraphPad, USA). One-way ANOVA followed by Post-hoc test (Bonferroni) was used to evaluate the level of statistical significance of clustering. The association between biofilm and proteinase production was determined by Pearson's correlation coefficient (r). Differences between proteinase/biofilm producers versus non producers were examined using Fisher's exact test. A P value < 0.05 was considered statistically significant.
Molecular typing of Candida parapsilosis isolates
AFLP was used to confirm correct species identification and to evaluate genetic variability within the selected 62 C. parapsilosis isolates. AFLP profiles obtained for C. parapsilosis consisted of 80 fragments ranging from 100 to 700 bases. Fragments larger than 700 bases were used as a control of DNA integrity. The number of monomorphic fragments was 62, which were common to all C. parapsilosis isolates. Therefore, these fragments were considered species specific and used for identification. Indeed, as shown in Figure 1A, which includes a wider panel of clinical isolates, this method allowed us to identify the presence of C. metapsilosis and C. tropicalis (CP542, CP534, CP557), which were excluded from this study. Identification of C. tropicalis and C. metapsilosis was performed by comparing AFLP profiles with those of 16 different fungal reference species [[16], data not shown].
Figure 1. AFLP patterns. (A) AFLP profiles obtained from the molecular screening of 48 putative Candida parapsilosis clinical isolates and reference strains ATCC 22019 (C. parapsilosis), ATCC 96139 (C. orthopsilosis) and ATCC 96143 (C. metapsilosis). In bold, isolates used in this study for genotyping and phenotyping isolated from Argentina (CP540-558) and Hungary (510-536). M 50-500 base molecular weigh standard. In italics, the non-parapsilosis isolates identified during the AFLP screening. (B) AFLP profiles of 34 C. parapsilosis strains isolated from Italy (CP1-CP502) and New Zealand (CP425-486). At the top of the figure, reference strains for C. metapsilosis (ATCC 96143) C. orthopsilosis (ATCC 96139) and C. parapsilosis (ATCC 22019) are included.
Figure 1 displays the AFLP profiles obtained from several C. parapsilosis isolates including those selected for the study and isolated from Argentina, Hungary (Figure 1A), Italy, and New Zealand (Figure 1B). When the presence/absence of fragments was the only parameter considered in AFLP analysis, very little genotypic diversity within the isolate collection was found (Figure 1A-B). In fact, the majority of AFLP markers included in the analysis (n = 80) were monomorphic, with only 18 polymorphic fragments. In agreement, UPGMA analysis indicated that all isolates grouped together, with a similarity index (SAB) higher than 0.96 (Figure 2A). A further level of analysis was performed to calculate a genetic distance between each isolate pair by the Pearson product-moment correlation coefficient (r), based on relative "intensity" of all fragments, which also takes into account the presence/absence of bands. Different band intensity was independent from DNA template concentration [data not shown; [28]], as expected, since AFLP is limited by primer concentration.
Figure 2. AFLP profile analysis. (A) UPGMA dendrogram based on presence/absence of AFLP fragments. (B) UPGMA dengrogram based on genetic distances derived from correlation coefficients including differences in relative band intensities. Geographical origin of isolates is indicated by I, Italy; RA, Argentina; NZ, New Zealand; and H, Hungary.
With this type of analysis, significant geographic clustering of the isolates was observed (Figure 2B). One-way ANOVA with Post-hoc test (Bonferroni) showed that all clustering above 0.04 (correlation coefficient of 0.96) were highly significant (P < 0.001). Reproducibility of the AFLP analysis was 97%, estimated by the average correlation among duplicated samples of reference C. parapsilosis strain (data not shown).
Drug susceptibility
All C. parapsilosis isolates were found to be susceptible to the antifungals included in the SensititreYeastOne® Y09 panel, with the exception of CP558 that displayed a dose-dependant susceptibility to fluconazole (MIC = 16 μg/ml). MIC values (μg/ml) were as follows: 5-flucytosine 0.06 ≤ MIC ≤ 0.25 (mean 0.127 ± 0.084 SD); posaconazole 0.008 ≤ MIC ≤ 0.5 (mean 0.069 ± 0.07 SD); voriconazole 0.008 ≤ MIC ≤ 0.5 (mean 0.037 ± 0.064 SD); itraconazole, 0.003 ≤ MIC ≤ 0.25 (mean 0.07 ± 0.036 SD); fluconazole, 0.125 ≤ MIC ≤ 16 (mean 1.8 ± 1.7 SD); amphotericin B, 0.125≤MIC≤ 1 (mean 0.44 ± 0.18 SD).
All C. parapsilosis isolates exhibited a reduced susceptibility to the echinocandin class, with the following MICs: anidulafungin, 0.5 ≤ MIC ≤ 2 (mean 1.32 ± 0.54 SD); micafungin, 0.5 ≤ MIC ≤ 2 (mean 1.17 ± 0.52 SD); caspofungin, 0.25 ≤ MIC ≤ 1 (mean 0.5 ± 0.22 SD).
All MIC values for echinocandin were ≤ 2 μg/ml (the defined cut-off value for susceptibility). However, caspofungin was the most active, with 85.5% of isolates showing MIC values ≤ 0.5 μg/ml.
Biofilm formation
To evaluate the effect of temperature on the formation of extra-cellular matrix, the production of biofilm was analyzed after 24 hour incubation at both 30°C and 37°C. As shown in Table 1, the majority of isolates produced biofilm (64.5%) following 24 hour incubation at 30°C, with similar results obtained after incubation at 37°C (64.3% of biofilm producers, data not shown). C. parapsilosis reference strain ATCC 22019 failed to produce biofilm at both temperatures tested. Statistically significant differences in the distribution of biofilm producers vs non producers were observed in strains isolated from different geographic regions. As shown in Figure 3A, the number of biofilm forming isolates (at 30°C) was significantly higher in isolates obtained from Hungary (P = 0.001) and Argentina (P = 0.011), compared with Italian strains, where a higher prevalence of non producers was found. The majority of isolates from New Zealand were biofilm producers. A similar trend was observed at 37°C (data not shown). When biofilm production was correlated with the anatomical origin of the samples, regardless of the geographical location, statistically significant differences in producers vs non producers could be observed between nail and blood isolates, with the latter encompassing a majority of biofilm producer strains, or between nail and cerebrospinal fluid samples (Figure 3B). Notably, all cerebrospinal fluid samples were isolated in Argentina. Again, results obtained at 30 and 37°C (data not shown) were similar. These experiments need to be confirmed with a wider range of isolates for each anatomical origin. Experimental variability was monitored by including a strong biofilm producer strain as a positive control in several experiments. Reproducibility experiments performed (n = 7) on separate days showed a mean absorbance of 0.348 ± 0.084 SD and a coefficient of variation of 24.1% [29]. The low standard deviation and a coefficient of variation of 24% indicated that good precision may be expected when using this method to estimate biofilm formation.
Figure 3. Biofilm production by C. parapsilosis. Biofilm production following 24 h incubation at 30°C in inducing medium by C. parapsilosis isolates obtained from different geographical areas (A) and different anatomical sites (B). Liquor stands for cerebrospinal fluid. Number of biofilm producing isolates (P) versus non producers (NP) were compared using Fisher's exact test. A P value < 0.05 was considered statistically significant. I = Italy, NZ = New Zealand, RA = Argentina, H = Hungary.
Proteinase secretion
Secretion of proteinase was measured as the proteolytic halo on solid BSA containing medium following 7 days incubation at 30°C. Most isolates were proteinase producers, with only 20 strains (33.9%) unable to hydrolyse BSA (Table 1). When the proteolytic activity was analysed in isolates obtained from different geographical regions an inverse trend was observed with respect to that obtained for biofilm production. In fact, a higher number of proteinase producers was found in Italy, and New Zealand, while they were significantly less represented in Hungary (P = 0.010 and 0.025, respectively, Figure 4A), where most biofilm producing strains were isolated. The analysis of protease production in isolates obtained from different body sites revealed no significant association between anatomical origin and production of this virulence factor (Figure 4B). The ATCC 22019 reference isolate showed no proteolytic activity (data not shown).
Figure 4. Proteinase secretion by C. parapsilosis. Proteinase secretion by C. parapsilosis isolates obtained from different geographical areas (A) and different anatomical origin (B). 'Liquor' refers to cerebrospinal fluid. Proteolytic activity was assessed on YCB-BSA agar after 7 days incubation at 30°C. Numbers of protease producing isolates (P) versus non producers (NP) were compared using Fisher's exact test. A P value < 0.05 was considered statistically significant. I = Italy, NZ = New Zealand, RA = Argentina, H = Hungary.
Univariate regression was applied to determine whether an association existed between the expression of the two virulence factors studied. As shown in Figure 5, a negative correlation between biofilm production and proteinase secretion by the C. parapsilosis isolates was observed (r = -0.483, P < 0.0001).
Figure 5. Correlation between biofilm and proteinase production. Negative correlation between biofilm production and proteinase secretion in Candida parapsilosis isolates (n = 62), as revealed by univariate regression analysis. Pearson's correlation coefficient (r) and P-value are indicated.
To date, no significant sequence variation has been described for Candida parapsilosis [30]. Therefore, this study was designed to provide further information on genotypic and phenotypic properties of this opportunistic fungal pathogen.
To evaluate the effect of different environments upon genetic variability C. parapsilosis isolates were selected to be representative of different geographical regions (Italy, Hungary, New Zealand, Argentina) and of different anatomical sites (blood, cerebrospinal fluid, mucosa, nail etc.). The EcoRI/HindIII enzyme combination used in the AFLP protocol was expected to produce a higher number of polymorphic bands since in C. metapsilosis band homoplasy was reduced with this combination and the average fragment length was larger than the one obtained with EcoRI/MseI [17]. Indeed the EcoRI/HindIII enzyme combination confirmed its higher discriminative power for C. parapsilosis and led to the identification of 20.7% of polymorphic fragments versus only 5% observed with EcoRI/MseI (data not shown). However, when genotype analysis was performed on the presence/absence of a band, the AFLP profiles obtained clearly indicated very high similarity, with all isolates grouped within a similarity index of 0.97.
This genetic variability is much lower than what we have observed for the species C. metapsilosis and C. orthopsilosis, for which we observed a greater number of polymorphic bands [16,17]. Our results are in agreement with the observation that the frequency of single nucleotide polymorphisms (SNPs) in C. parapsilosis is 30 to 70 fold lower than in other Candida species [30]. The low level of variability found suggests a clonal or selfing strategy of reproduction, supporting the hypothesis of a successful species recently emerged as a genetically homogeneous population diverged from a common ancestor [31]. It is not possible to determine whether the low variability found is due to a real absence of sexual recombination, as supported by the finding of a defective mating-like type locus [30,32], since the very limited number of variable fragments hinders a statistical analysis to prove/disprove the existence of recombination within this species, as previously performed for C. orthopsilosis and C. metapsilosis [16,17]. Interestingly, a recent manuscript by Sabino and colleagues [33] reports a high degree of polymorphisms by microsatellite analysis in C. parapsilosis, with 192 different genotypes found among 233 isolates, based on 4 hyper variable loci. This is remarkable, considering that the majority of the literature points towards limited genetic variability in this species. The hypervariability found can provide an excellent tool to discriminate between isolates in outbreak investigations. However, it does not seem to be useful for genetic relatedness studies on larger time scale or on population structure [33].
When the genetic distance between each isolate pair was calculated using the Pearson's coefficient, which takes into account both the presence/absence of bands and their relative "intensity", significant geographic clustering of the isolates was obtained (P < 0.001). This coefficient has been used as an index of genetic distance and has been previously reported in AFLP analysis of bacteria [34,35] and Candida species [36]. Candida fingerprinting techniques such as RFLP with species specific probes, RAPD, karyotyping also produce band patterns which differ in band mobility and intensity. In this respect, genotyping with AFLP gives rise to a much more complex pattern, composed by a larger number of bands, which can be compared by mobility and intensity [37]. The accuracy of the Pearson's coefficient is also dependent on the number of fragments included in the comparison. Thus, generating over 80 fragments with a single enzyme/primer combination, AFLP seems to be a suitable tool to perform this kind of analysis [37].
In this context, it is interesting to speculate what causes the variation in the relative band intensities. Karyotypes differing in band mobility and intensity have already been described for C. parapsilosis and other Candida species [[38], data not shown] and Butler and co-authors showed that C. albicans can be partially hemizygous [30]. The role that ploidy plays in C. parapsilosis genetic variability is a phenomenon already described. In fact, it was shown that its nuclear size ranges from 57% to 86% from its estimated diploid size [30,39]. We assume that one haploid complete set of the genome (50%) is always present in the isolates but what the remaining 7 to 36% of the DNA actually represents remains unknown. Whether this represents between 7 to 36% of one homologous set and/or whether these are DNA sequences present in variable copy numbers is still to be determined.
Using AFLP with the enzyme combinations EcoRI, HpaII, and MspI, we have noted that in C. parapsilosis, methylation of cytidine occurs. It was also observed that this methylation was variable in different isolates (data not shown). Variable DNA methylation of restriction sites might be responsible for different AFLP profiles, including the observed differences in relative intensities of AFLP bands found in this study with the enzymes HindIII and EcoRI.
For what concerns phenotypic traits, drug susceptibility tests showed that all isolates were susceptible to the antifungals tested, with the exception of one fluconazole dose-dependant susceptible isolate. Regardless of the geographical or anatomical origin, a reduced susceptibility to echinocandins was observed for all isolates, confirming what has already been described for this species [40]. It has been suggested that this phenotype is due to a naturally occurring Proline to Alanine amino acid change (P660A) in the glucan synthase enzyme Fks1p [40]. However, MIC values were all ≤ 2 mg/ml, the accepted breakpoint for echinocandins against Candida species [26,27].
Since this fungal pathogen is able to colonise body sites with different core temperatures, we examined whether biofilm formation was influenced by incubation at 30 or 37°C. The results obtained indicated that this parameter does not significantly alter the ability to produce biofilm in vitro, with minor differences in the quantity of the extracellular matrix produced at different temperatures. Interestingly, biofilm production was linked to both geographical and anatomical origin of isolates; indeed, Argentinian or Hungarian isolates produced significantly more biofilm than Italian strains. To date we do not have an explanation to justify the higher biofilm production that was observed in Hungarian isolates. The majority of these high biofilm producers came from surgery or intensive care units, where catheter related infections with biofilm producer isolates are more commonly found. Of note, even though the analysis was performed on a limited number of isolates, blood and cerebrospinal fluid isolates were found to be more frequently biofilm producers than strains isolated from nails. These findings need to be confirmed by comparing a wider set of isolates for each anatomical site of origin.
The majority of C. parapsilosis isolates (66.1%) produced proteinase in vitro. In contrast to what was observed for biofilm production, proteinase producers were mostly detected in Italy and New Zealand. Interestingly, a statistically significant inverse correlation was found between proteolytic activity and the ability to form biofilm, independent of the geographical/anatomical origin of isolates. Indeed, this finding has also been described for Staphylococcus aureus [41], where extracellular proteases make a significant contribution to a biofilm deficient phenotype of an S. aureus mutant, as shown by the addition of proteinase inhibitors to biofilm formation assay [41]. In addition, Boles and Horswill [42] demonstrated through genetic analysis that an S. aureus double mutant in a metalloprotease and serine protease, displaying minimal extracellular protease activity, was improved in biofilm formation, and had a strongly attenuated detachment phenotype. These findings demonstrate that the S. aureus dispersal mechanism from consolidated biofilm requires extracellular protease activity. Recently, the existence of a new pathway has been demonstrated, controlling protein-mediated biofilm formation in which different global regulators modulate biofilm formation by controlling the expression of S. aureus extracellular proteases [43]. Therefore, in analogy to what is described for S. aureus, we hypothesise that the negative impact of extracellular proteases on biofilm formation is multifactorial, potentially promoting detachment and release from a mature biofilm, via degradation of C. parapsilosis adhesins and/or extracellular matrix components.
Overall, these results confirm previous evidence that Candida parapsilosis is characterised by a limited DNA sequence variability, even when considering isolates collected from distant geographical regions. The fact that phenotypic properties were found to significantly differ in strains isolated from various geographical regions suggests that other mechanisms such as epigenetic modifications may be used by this yeast to adapt to environmental changes.
Authors' contributions
AT designed the study with LAMH, performed phenotypical analysis and drafted the manuscript; LAMH conceived the study with AT, performed AFLP analysis and wrote the manuscript; SM participated in the drug susceptibility assays; LM has made substantial contribution to acquisition of data and critically revised the manuscript. SS participated in the study coordination and has made substantive contribution to data analysis; MC participated in the study design and has given the final approval to the version to be published.
All authors have read and approved the final version of the manuscript.
This study was supported by the research grant no. 2005068754 from the Italian Ministero dell'Istruzione, dell'Università e della Ricerca and by Merck & Co. Inc. We are grateful to Prof Giulia Morace, Dr Arlo Upton and Dr Marisa Biasoli who provided us with isolates. We also thank Dr Colin G. Egan for revising the manuscript for English language.
Lockhart SR, Messer SA, Pfaller MA, Diekema DJ: Geographic distribution and antifungal susceptibility of the newly described species Candida orthopsilosis and Candida metapsilosis in comparison to the closely related species Candida parapsilosis.
J Clin Microbiol 2008, 46:2659-2664. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Pfaller MA, Diekema DJ, Gibbs DL, Newell VA, Ng KP, Colombo A, Finquelievich J, Barnes R, Wadula J, Global Anifungal surveillance Group: Geographic and temporal trends in isolation and antifungal susceptibility of Candida parapsilosis: a global assessment from the ARTEMIS DISK Antifungal Surveillance Program, 2001 to 2005.
J Clin Microbiol 2008, 46:842-849. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Almirante B, Rodriguez D, Cuenca-Estrella M, Almela M, Sanchez F, Ayats J, Alonso-Tarres C, Rodriguez-Tudela L, Pahissa A: Epidemiology, risk factors, and prognosis of Candida parapsilosis bloodstream infections: case-control population-based surveillance study of patients in Barcelona, Spain, from 2002 to 2003.
J Clin Microbiol 2006, 44:1681-1685. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Pfaller MA, Diekema DJ: Epidemiology of invasive candidiasis: a persistent public health problem.
Clin Microbiol Rev 2007, 20:133-163. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Colombo AL, Guimaraes T, Silva LR, de Almeida Monfardini LP, Cunha AK, Rady P, Alves T, Rosas RC: Prospective observational study of candidemia in Sao Paulo, Brazil: incidence rate, epidemiology, and predictors of mortality.
Infect Control Hosp Epidemiol 2007, 28:570-576. PubMed Abstract | Publisher Full Text
Trofa D, Gácser A, Nosanchuk JD: Candida parapsilosis, an emerging pathogen.
Clin Microbiol Rev 2008, 21:606-625. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Levin AS, Costa SF, Mussi NS, Bass M, Sinto SI, Machado C, Geiger G, Villares MC, Schreiber Z, Barone A, Branchini ML: Candida parapsilosis fungemia associated with implantable and semi implantable control venous catheters and the hands of healthcare workers.
Diagn Microbiol Infect Dis 1998, 30:243-249. PubMed Abstract | Publisher Full Text
Lupetti A, Tavanti A, Davini P, Ghelardi E, Corsini V, Merusi I, Boldrini A, Campa M, Senesi S: Horizontal transmission of Candida parapsilosis candidemia in a neonatal intensive care unit.
J Clin Microbiol 2002, 40:2363-2369. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Fridkin SK, Kaufman D, Edwards JR, Shetty S, Horan T, The National Nosocomial Infection Surveillance System Hospitals: Changing incidence of Candida bloodstream infections among NICU patients in the United States:1995-2004.
Pediatrics 2006, 117:1680-1687. PubMed Abstract | Publisher Full Text
Kuhn DM, Chandra J, Mukherjee PK, Ghannoum MA: Comparison of biofilms formed by Candida albicans and Candida parapsilosis on bioprosthetic surfaces.
Infect Immun 2002, 70:878-888. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Shin JH, Kee SJ, Shin MG, Kim SH, Shin DH, Lee SK, Suh SP, Ryang DW: Biofilm production by isolates of Candida species recovered from nonneutropenic patients: comparison of bloodstream isolates with isolates from other sources.
J Clin Microbiol 2002, 40:1244-1248. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Tumbarello M, Posteraro B, Trecarichi EM, Fiori B, Rossi M, Porta R, de Gaetano Donati K, La Sorda M, Spanu T, Fadda G, Cauda R, Sanguinetti M: Biofilm production by Candida species and inadequate antifungal therapy as predictors of mortality for patients with candidemia.
J Clin Microbiol 2007, 45:1843-1850. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Vrioni G, Matsiota-Bernard P: Molecular typing of Candida isolates from patients hospitalized in an intensive care unit.
J Infect 2001, 42:50-56. PubMed Abstract | Publisher Full Text
Kuhn DM, Mukherjee PK, Clark TA, Pujol C, Chandra J, Hajjeh RA, Warnock DW, Soll DR, Ghannoum MA: Candida parapsilosis characterization in an outbreak setting.
Emerg Infect Dis 2004, 10:1074-1081. PubMed Abstract | Publisher Full Text
Tavanti A, Davidson AD, Gow NAR, Maiden MCJ, Odds FC: Candida orthopsilosis and Candida metapsilosis spp. nov. to replace Candida parapsilosis groups II and III.
J Clin Microbiol 2005, 43:284-292. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Tavanti A, Hensgens LA, Ghelardi E, Campa M, Senesi S: Genotyping of Candida orthopsilosis clinical isolates by amplification fragment length polymorphism reveals genetic diversity among independent isolates and strain maintenance within patients.
J Clin Microbiol 2007, 45:1455-1462. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Hensgens LA, Tavanti A, Mogavero S, Ghelardi E, Senesi S: AFLP genotyping of Candida metapsilosis clinical isolates: evidence for recombination.
Fungal Genet Biol 2009, 46:750-758. PubMed Abstract | Publisher Full Text
Dagdeviren M, Cerikcioglu N, Karavus M: Acid proteinase, phospholipase and adherence properties of Candida parapsilosis strains isolated from clinical specimens of hospitalises patients.
Mycoses 2005, 48:321-326. PubMed Abstract | Publisher Full Text
Borst A, Theelen B, Reinders E, Boekhout T, Fluit AC, Savelkoul PHM: Use of amplified fragment length polymorphism analysis to identify medically important Candida species, including C. dubliniensis.
J Clin Microbiol 2003, 41:1357-1362. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Barchiesi F, Spreghini E, Tomassetti S, Della Vittoria A, Arzeni D, Manso E, Scalise G: Effects of caspofungin against Candida guilliermondii and Candida parapsilosis.
Antimicrob Agents Chemother 2006, 50:2719-2727. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Perlin DS: Resistance to echinocandin-class antifungal drugs.
Drug Resist Updat 2007, 10:121-130. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Kalinowski ST: How well do evolutionary trees describe genetic relationships between populations.
Heredity 2009, 102:506-513. PubMed Abstract | Publisher Full Text
Hampl V, Pavlíček A, Flegr J: Construction and bootstrap analysis of DNA fingerprinting-based phylogenetic trees with the freeware program Freetree: application to trichomonad parasites.
Int J Syst Evol Microbiol 2001, 51:731-735. PubMed Abstract | Publisher Full Text
Page RDM: and application to display phylogenetic trees on personal computers.
Comp Appl Biosci 1996, 12:357-358. PubMed Abstract
Rüchel R, Tegeler R, Trost M: A comparison of secretory proteinases from different strains of Candida albicans.
Sabouraudia 1982, 20:233-244. PubMed Abstract
CLSI (a): Reference method for broth dilution antifungal susceptibility testing of yeasts; approved standard-Third Edition. In CLSI document M27-A3. Wayne, PA: Clinical and Laboratory Standards Institute; 2008.
CLSI (b): Reference method for broth dilution antifungal susceptibility testing of yeasts; third informational supplement. In CLSI document M27-S3. Wayne, PA: Clinical and Laboratory Standards Institute; 2008.
Bensch S, Akesson M: Ten years AFLP in Ecology and evolution: why so few animals?
Mol Ecol 2005, 14:2899-2914. PubMed Abstract | Publisher Full Text
Riefler RG, Ahlfeld DP, Smets BF: Respirometric Assay for Biofilm KineticsEstimation: Parameter Identifiability and Retrievability.
Biotech and Bioeng 1998, 57:35-45. Publisher Full Text
Butler G, Rasmussen M, Lin MF, Santos MA, Sakthikumar S, et al.: Evolution of pathogenicity and sexual reproduction in eight Candida genomes.
Nature 2009, 459:657-662. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Nosek J, Holesova Z, Kosa P, Gacser A, Tomaska L: Biology and genetics of the pathogenic yeast Candida parapsilosis.
Curr Genet 2009, 55:49-509. Publisher Full Text
Logue ME, Wong S, Wolfe KH, Butler G: A genome sequence survey shows that the pathogenic yeast Candida parapsilosis has a defective MTLa1 allele at its mating type locus.
Eukaryot Cell 2005, 4:1009-1017. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Sabino R, Sampaio P, Rosado L, Stevens DA, Clemons KV, Pais C: New polymorphic microsatellite markers able to distinguish among Candida parapsilosis sensu stricto isolates.
J Clin Microbiol 2010, 48:1677-82. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Hänninen ML, Perko-Mäkelä P, Rautelin H, Duim B, Wagenaar JA: Genomic relatedness within five common Finnish Campylobacter jejuni pulsed-field gel electrophoresis genotypes studied by amplified fragment length polymorphism analysis, ribotyping, and serotyping.
Appl Environ Microbiol 2001, 67:1581-1586. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
van Eldere J, Janssen P, Hoefnagels-Schuermans A, van Lierde S, Peetermans WE: Amplified-fragment length polymorphism analysis versus macro-restriction fragment analysis for molecular typing of Streptococcus pneumoniae isolates.
J Clin Microbiol 1999, 37:2053-2057. PubMed Abstract | PubMed Central Full Text
Lopes MM, Silva D, Freitas G, Tenreiro R: Simultaneous identification and typing of Candida species by MSP-PCR and AFLP: study of clinical isolates from a Portoguese pediatric hospital.
Savelkoul PH, Aarts HJ, de Haas J, Dijkshoorn L, Duim B, Otsen M, Rademaker JL, Schouls L, Lenstra JA: Amplified-fragment length polymorphism analysis: the state of an art.
J Clin Microbiol 1999, 37:3083-3091. PubMed Abstract | PubMed Central Full Text
Lott TJ, Kuykendall RJ, Welbel SF, Pramanik A, Laser BA: Genomic heterogeneity in the yeast Candida parapsilosis.
Curr Genet 1993, 23:463-467. PubMed Abstract | Publisher Full Text
Fundyga RE, Kuykendall RJ, Lee-Yang W, Lott TJ: Evidence for aneuploidy and recombination in the human commensal yeast Candida parapsilosis.
Garcia-Effron G, Katiyar SK, Park S, Edlind TD, Perlin DS: A naturally occurring proline-to-alanine amino acid change in Fks1p in Candida parapsilosis, Candida orthopsilosis, and Candida metapsilosis accounts for reduced echinocandin susceptibility.
Antimicrob Agents Chemother 2008, 52:2305-2312. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Tsang LH, Cassat JE, Shaw LN, Beenken KE, Smeltzer MS: Factors contributing to the biofilm-deficient phenotype of Staphylococcus aureus sarA mutants.
PLoS One 2008, 3:e3361. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Boles BR, Horswill AR: Agr-mediated dispersal of Staphylococcus aureus biofilms.
PLoS Pathog 2008, 4:e1000052. PubMed Abstract | Publisher Full Text | PubMed Central Full Text
Martí M, Trotonda MP, Tormo-Más MA, Vergara-Irigaray M, Cheung AL, Lasa I, Penadés JR: Extracellular proteases inhibit protein-dependent biofilm formation in Staphylococcus aureus.
Microb Infect 2010, 12:55-64. Publisher Full Text


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NOTE: Post 15, below is a Direct Link to Compare Intel Processors: Laptop, Desktop, Server That has turned out to be helpful.
I would like to build/spec out a computer to handle Poser 9 and above. Cost is an issue and I'm looking for a guide as to where I really should not skimp.
My main objective for the new system would be to have it minimize Render times. I would also like to create larger models that don't use up the available memory leaving nothing for the renderer.
I would like to know what people are currently using and what they think the critical components for fast graphics are.
What do you consider state of the art and what have you all experienced with newer systems?? What would you change?
Any idea what kind of rendering time improvements I can reasonably expect over the older XP system I have now?
I don't know much about Poser 9 system details. I am told it can access 64 bit for working , but that I can't efficiently render with it no matter what new computer I build [edit: no: runs 32 bit only; can run 32 bit under Windows 7] . If that is the case, what do 'normal' 3D programs (or Poser 2012) run best on. Does Poser 9 access more than one core if you have a multi-core system? [edit: no. 32 bit only]
Here are my basic question areas with the techy, but feel free to ignore them to tell me what you think. Also, I may not be even asking the pertinent questions:
(1) Intel or AMD? How many cores? Core memory size?
(2) If Intel I5 or I7?
(3) Graphics card (GPU). How important? How much dedicated memory makes a difference. Any particular brand/vendor?
(4) Are busses important? [Ivy or Sandy].
I welcome input from people who use other 3D Graphics software, too. Daz users!!! C4, Rhino, Maya, Lightwave, Autodesk and Blender. Shade, Hexagon…. What am I leaving out?
Any places I should look (magazine articles you may have seen, threads, forums?

subset: commoncrawl


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August 18, 2012 12:00:00 AM
A 32-year-old woman has pleaded guilty to causing the death of two people at Sovi Bay on Thursday night.
The accused, Vijenti Banda appeared before Magistrate Peni Dalituicama at the Lautoka Magistrates Court pleaded guilty to two counts of dangerous driving causing death and one count of dangerous driving occasioning grievous bodily harm.
Police spokeswoman Ana Naisoro said Banda did not want legal representation.
Banda was ordered to surrender her passport and not to re-offend and was bailed for the sum of $1,500 and to report to the Nadi Police Station today and tomorrow then to the Sigatoka Police Station on Monday morning.
The case has been adjourned to August 29, 2012 for facts and sentencing at the Sigatoka Magistrates Court.
In another case, Etonia Bolabasaga, 41, also appeared before Dalituicama charged with one count of Dangerous Driving Occasioning Death.
Naisoro said the accused was represented by legal counsel and deferred his plea after the charges were read and explained in court.
Bolabasaga was bailed for $1,500 and ordered not to obtain a passport, not to reoffend, surrender driving license and to report to the Lautoka Police Station every Monday, Wednesday and Friday between 6am to 6pm.
The case has been deferred to September 7, 2012 for mention.
Naisoro said prosecution has been given 14 days to serve disclosures.
The accused is alleged to have hit a man at Navutu Road, Lautoka.
By Ropate Valemei
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Multicolored autism awareness ribbon with interlocking puzzle pieces.
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This scholarship was established in 2000 by Allan Glenn based upon his experience of support and assistance during his years as a student athletic trainer at the University. It also honors the memory of his father, Mr. Raymond Hagmeier, who devoted many years to officiating baseball at the University.
This scholarship is designed to provide an opportunity for an athletic training student to have a portion of his/her education funded. The recipent of this scholarship must maintain good standing in the athletic training education program. The scholarship is awarded without regard to financial need, QPA, and county/state of residence. Two scholarships will be awarded, one for a junior and one for a senior level student. The award will be divided equally between the fall and spring semester. The scholarship is renewable as long as the student maintains good standing in the academic program.
Pictured left to right: back row (L-R), Mr. Allan Glenn and Jason Bittle; front row (L-R), Hillary Helms and Tashara Ardner


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New Orleans, La. – The Big Easy Foundation honors outstanding achievement in Classical Music, Opera and Dance today at the 16th Annual Tribute to the Classical Arts. Dr. John Ware, Xavier University music professor and conductor of the New Orleans Black Chorale receives the 2010 Big Easy Lifetime Achievement Award.


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2:45 pm
Thu December 19, 2013
Do Recommendations From White House Panel On NSA Go Far Enough?
By editor
Originally published on Thu December 19, 2013 3:42 pm
A review panel convened by the White House released its report on surveillance by the National Security Agency Wednesday. Among concerns the task force has are NSA practices that some say deliberately weaken America's cyber security. The Electronic Frontier Foundation has been critical about surveillance activities, and Mark Rumold, staff attorney for the organization, tells Audie Cornish about the pros and cons of the panel's recommendations.


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Protecting your site and your clients
Web Security
Learn more about protecting your website with NoScript, ModSecurity, and Site Security Policy.
Like many security issues, the World Wide Web presents two very different sets of problems with some very different solutions. On the one side, most of us use a web browser on a regular basis and want to prevent our web clients from running an attacker's code, letting them take over our machine. On the other side are web servers, which you don't want to see compromised, under constant attack (XSS, SQL injection, etc.). So what's the answer? Well, there is no single answer. You need to take steps to protect both the clients and the servers because no matter how security conscious you are, you will interact with servers or clients that are less secure.
JavaScript and NoScript
Securing web clients against attacks is relatively simple; however, some websites might not work properly. Disabling JavaScript entirely is one option, but many sites now rely on JavaScript to present content, forms, and so on.
A more fine-grained approach is available with the NoScript plugin for Firefox [1]. The default is to block JavaScript execution, and then you can choose to allow JavaScript to run temporarily or permanently. Or, you can permanently mark a site as untrusted to prevent any JavaScript from ever being executed from that site (Figure 1).
The major downside to this plugin is that you need to pay attention to the information bar that pops up at the bottom of the screen when JavaScript is blocked (Figure 2) and decide whether or not to allow it. If you don't, you will find yourself, as I have, staring at a website wondering why it is mostly blank, or why an online form isn't working properly.
Additionally, NoScript has some basic cross-site scripting protection – URLs with characters such as ">" in them will generate a warning and give the user a chance to block their loading.
Securing the Server
As a server admin, you can't force clients to be secure, but you can protect your own server and web-based applications from attack. Protecting your server can also prevent broken clients or users who have visited hostile sites from taking actions that might harm their accounts or the data hosted on your site – for example, from a cross-site scripting attack that interacts with the user's account to change the password on their account at your site.
Apache ModSecurity
Like many security projects, ModSecurity started out as an open source project, licensed under the GPL v2 and aimed at adding a layer of security to the Apache web server [2]. The project appears to have been commercialized successfully; however, like many open source security applications, free versions are still available.
The main benefit of ModSecurity is that you can use it to provide security for any application running on your system. On the downside, you must be able to insert a custom module into Apache – meaning you need to have control over the server – and have enough CPU power to handle the additional processing required by this module, which can be significant. The ModSecurity module allows requests to the server to be examined at various stages in the process: when the request header is first processed, when the request body is processed, when the response headers are created, when the response body is processed, and at the logging phase.
Another advantage of ModSecurity is that it supports Perl-Compatible Regular Expressions (PCRE), and the rules it supports can also trigger a variety of actions, including to allow or block a request, to drop the connection by sending a FIN packet, or to execute an external program. For example, this allows site admins to filter out characters such as "<" and ">" from requests – a likely indicator of a cross-site scripting attack – or to look for personal information such as 12-digit credit card number strings within outgoing requests (e.g., triggered by an SQL injection attack) and block that data from being served to the attacker. See the "Example Rule" box.
As you can imagine, all this power and flexibility comes at the cost of complexity; however, this is alleviated because a powerful default rule set has been made available for free (licensed under the GPL v2), which can be used as a starting point for most sites.
Example Rule
A simplistic example to detect and block any 12-digit number in outgoing web pages:
SecRule RESPONSE_BODY "[0-9]{12}" \ "phase:4,t:none,ctl:auditLogParts=+E,deny,log,auditlog,status:500,msg:'a 12 digit number',id:'1',tag:'LEAKAGE/ERRORS',severity:'1'"
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Occupational Safety Degrees by Degree Level
Learn about occupational safety degree programs by level. Discover the courses that are included in each program and review a list of potential careers for graduates. See the employment outlook statistics for occupational health and safety specialists.
Essential Information
Depending on their previous education, individuals who want to build careers in occupational safety could apply to bachelor's or master's programs. Once enrolled in a 4-year bachelor's degree program in occupational safety, students learn to monitor workplace safety conditions and properly operate equipment. Bachelor's-level students might participate in internships for hands-on experience. A 2-year master's degree program in occupational safety provides advanced education for experienced professionals who want to increase their knowledge of safety engineering, fire protection and toxicology.
Bachelor of Science in Occupational Safety
Occupational safety professionals protect the public from potentially unsafe working conditions. With safety protocols in place for practically every industry, occupational safety specialists need to be qualified and trained to visit sites to find code violations, understand chemicals, analyze an accident and know how to handle a situation involving hazardous materials.
A bachelor's degree program in occupational safety prepares students for real world experiences that require them to anticipate, solve and monitor conditions that are unfit or illegal for workers. Students learn management and technical skills, such as how to operate machinery to test its usability and how to prevent workplace fires. High school graduates are eligible to enroll in a bachelor's degree in occupational health and safety program.
Program Coursework
Students in the bachelor's degree in occupational safety program are exposed to a range of topics, including how to use protective equipment. Often, students are required to complete an internship in occupational health and safety and learn about workers' compensation. Typical courses include:
- Accident prevention
- Safety engineering
- Environmental issues
- Safety, health and the law
- Employment relations
- Occupational Safety and Health Administration (OSHA) standards
Popular Career Options
Individuals with a bachelor's degree in occupational health and safety have the knowledge to work in a variety of fields, from insurance to business, or for federal, state or local government. Popular careers include:
- Loss prevention specialist
- Environmental protection officer
- Occupational health and safety inspector
- Ergonomist
- Industrial hygienist
Certification Information
With a bachelor's degree in occupational safety, graduates may apply for an optional professional certification. The Board of Certified Safety Professionals offers a Certified Safety Professional designation, which tests knowledge of the safety standards via a computer-based examination (www.bcsp.com). Recertification requirements need to be met every five years.
Master's in Occupational Safety
While undergraduate degree programs serve as introductions to the field of occupational safety and health, the master's degree program focuses on preventing and solving safety and health dangers. Students in the graduate degree program are often already experienced in the field and enroll in the program to advance in their career opportunities and strengthen their leadership skills. Once in the field, graduates may train workers and employers, implement new safety and health practices for violators, perform on-site inspections and design emergency response plans.
Admission into the master's degree in occupational safety program requires a bachelor's degree in a related field, like science or engineering.
Program Coursework
The master's degree in occupational safety curriculum concentrates on the techniques to make decisions, how to reduce workplace violations, conduct audits and promote workplace safety. Students develop the skills needed to be a leader in the occupational safety industry through classroom assignments and case studies on topics like:
- Advanced toxicology
- Research methods
- Accident prevention
- Safety engineering
- Critical thinking
- Occupational ergonomics
- Fire protection
Employment Outlook and Salary Info
According to the U.S. Bureau of Labor Statistics (BLS), a projected 9% increase in employment of occupational health and safety specialists was expected from 2010-2020; the majority of these jobs were anticipated to be for loss prevention professionals (www.bls.gov).
The same source reported that in May 2011, there were 57,950 occupational safety and health specialists working in the United States. The median salary for these professionals was $31.86 per hour, or $66,270 annually.
Continuing Education and Professional Certification
Individuals seeking further education can complete a doctorate in occupational health and safety, although these degrees aren't incredibly common. The Board of Certified Safety Professionals offers a Graduate Safety Professional for master's degree graduates who apply for the qualifying examination within three years of graduation.
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Good for tank fish too
posted 5 years ago
I was told many years ago to use a small amount in with my livebearers as they like their water a bit brackish so I continued to do it with my pond wish also.
Pros: Does the job
Cons: None
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Originally written on Razzball Football | Last updated 11/15/14
For yet another season, Razzball will be interviewing local NFL beat writers for some in-depth actual football knowledge to sway some insight in our fantasy football knowledge. Keep your eye out for an interview for every NFL team through the summer and check out each one on the "2012 Fantasy Football Team Previews" link. This installment comes courtesy of David Wyatt from New York Jets blog Gang Green Nation: 1) Mark Sanchez set two career highs last year with 32 total touchdowns and 26 total turnovers. Obviously, one of them was really a career low but you get what I'm saying. Though the numbers weren't completely terrible, he did regress from his 2010 campaign in ways. Now with Tim Tebow on board, are there any questions about who the number one quarterback in Jets camp is and how long it will stay that way? Last year we tried to put too much on Sanchez, he'll never be a Quarterback who should throw the football 50 times a game. The Verbiage in the Schottenheimer offen...
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Female impersonation need say nothing about sexual identity, but it has for a long time been almost an institutionalized aspect of gay male culture.
Although sparse in images documenting the gay community, pre-Stonewall gay male photography blurs the boundaries between art, erotica, and social history.
Given the historic stigma around making, circulating, and possessing overtly homoerotic images, the visual arts have been especially important for providing a socially sanctioned arena for depicting the naked male body and suggesting homoerotic desire.
Independent films that aggressively assert homosexual identity and queer culture, the New Queer Cinema can be seen as the culmination of several developments in American cinema.
Renowned photographer, teacher, critic, editor, and curator, Minor White created some of the most interesting photographs of male nudes of the second half of the twentieth century, but did not exhibit them for fear of scandal.
The first international fashion superstar, Halston dressed and befriended some of America's most glamorous women.
An artistic movement that grew out of Dadaism and flourished in Europe shortly after World War I, Surrealism embraced the idea that art was an expression of the subconscious.
Film, stage, and television actor Paul Winfield was openly gay in his private life, but maintained public silence about his homosexuality.
Detail from an untitled work by Tom of Finland. © Tom of Finland® Foundation.
An exhibition of the art of Tom of Finland (Touko Laaksonen, 1920-1991), a pioneer in twentieth-century gay male erotic art, will be presented by the World Erotic Art Museum in Miami Beach from April 14 through July 31, 2012. The show, entitled "TOM OF FINLAND: Male Masterworks" will feature twenty paintings and drawings, some of which have never before been exhibited. The exhibition is scheduled to coincide with Pride season in Miami Beach.
Curator Volker Morlock will give a talk at the exhibition's opening, which will be held on Saturday, April 14, 2012 from 7:00 pm until midnight at the museum. Morlock has arranged the works on display chronologically to allow visitors to see the progression of the artist's style and the evolution of his choices of subjects.
Untitled, 1972 (from "In the Park") © Tom of Finland® Foundation, 1972.
In addition to the art works on display, the exhibition will include a new video tribute to the artist as well as vintage memorabilia.
Untitled ("Sailor's Dream"), 1959. © Tom of Finland® Foundation, 1959.
Glbtq contributor Ken Gonzalez-Day calls Tom of Finland an iconic artist who "has created some of the most indelible (and erotic) images of twentieth-century gay life and has fueled the fantasies of generations of gay men." His work retains its relevance and erotic punch today.
Untitled (From the "The Tattooed Sailor" series), 1962. © Tom of Finland® Foundation, 1962.
For more information about the exhibition, visit The Tom of Finland Foundation or The World Erotic Art Museum.
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Publisher's Description+
From GARMIN Corporation:
Garmin USB GPS
The Global Positioning System (GPS) is a satellite-based navigation system made up of a network of 24 satellites placed into orbit by the U.S. Department of Defense. GPS was originally intended for military applications, but in the 1980s, the government made the system available for civilian use. GPS works in any weather conditions, anywhere in the world, 24 hours a day. There are no subscription fees or setup charges to use GPS.
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Author has written 3 stories for Supernatural.
I'm back! But it will be slow progress as I work a full time job and a part time job, but I will post as I complete chapters!
General Info:
Name: Sarah
Gender: Female
Age: 19
Likes: Animals, Fiction, Supernatural, Books, School, criticism (it makes me a better writer)
Dislikes: People who think they are better than anyone else, animal abusers, immaturity, people who are afraid to admit their feelings
(i.e. if your scared admit it. It takes more courage to admit you are scared than to pretend to be brave.)
Random Stuff:
For those who wish they could change the past: Remember what happened then made you who you are today!
If you could would you either travel to the future or the past? Neither. The past only hold events that if changed would change you. The future only holds answers to questions you have yet to ask and events you have yet to experience.
Live your life! You only have one.
There is no magical age where one becomes an adult. There is no amount of years that can grant maturity. Only learning can grant those two things. Learning from your mistakes. Learning from the mistakes of others. Learning from hardships. Learning from the good times. Life is full of lessons and only learning from them will make you a mature adult.
Stereotypes: A word for "I'm too lazy to actually get to know a person so I'll create a bunch of categories and everytime I meet a person I'll just take five seconds then toss them into one of them."
You are the only one who can control your happiness. No one has the right to try to make you unhappy.
Depression is an illness that affects your state of mind, not a general illness or even a mental disease. Therefore you can't treat depression through medication alone. To truly get over depression you have to find out what the cause of your depression is and work through that. Anti-depressants can only aid you while you search for the true cure. Therapists can also help you work through your depression. I personally recommend the latter, as it's much healthier for you than pills.
I live in America and we are supposed to be free. But all around me I see people using their rights to strip others of theirs. I find this behavior disgusting. Don't use your right to freedom of religion to take away others' (Gay Rights). Don't use your right to freedom of religion to take my equal rights away (Abortions are covered under womens rights). Don't use your right to religious freedom to control our country (Church Separate from State). Above all keep your damn religious beliefs to yourself. I don't go around telling everyone that you're going to hell every time you take a deer's life or litter or don't recycle and those are against MY religious beliefs. Keep your opinions to yourself. You can't control everyone. Newsflash: If you're so worried about going to hell then don't be gay and don't get an abortion. No one is making you do anything. All we are asking is you let others live their lives they way they have A RIGHT TO LIVE IN THE LAND OF THE FREE; NOT THE LAND OF THE CHRISTIANS.
I get my inspiration from the books I read and my life.
I've lived a pretty difficult life. Because of that I know the harder side of life, but that just adds a better quality to my story because I know hard times and so I can write about hard times. And because of some pretty amazing friends I have the pleasure of knowing the sweeter side too.
Every book I read gives me some sort of inspiration, so some of my favorites are: Night World by LJ Smith; Twilight by Stephenie Meyers; Most books by Nora Roberts; Infinite Days (unsure who its by); Mortal Instruments by Cassandra Clare; Harry Potter by J.K. Rowling; The Hunger Games; Inheritance Cycle (Better known as Eragon) by Christopher Poalini; Temperance Brennan Series by Kathy Reichs; All series by Charlaine Harris; Blue Bloods by Melissa de La Cruz; Midnight Breed Series by Laura Adrian; Vampire Academy by Richelle Mead; Bloodlines by Richelle Mead and many many more!!!
My Forum:
Name: Triple Moon
URL: http://www.fictionpress.com/forum/Triple-Moon/6929/
Description: You can request reviews, chat with people, discuss/develop characters, and hopefully find some pretty awesome stories.
Beta Reader:
I am no longer accepting beta requests.
For those who do not know a beta reader is a person who reads a work of fiction with a critical eye, with the aim of improving grammar, spelling, characterization, and general style of a story prior to its release to the general public. But I will also look at stories that have been posted.
My Stories:
Ash and The Wolf; It's been over a year since Dominic found out she was actually Ash; a powerful half-vampire; half-witch. She's just started a new school, full of people just like her; supernaturals. She thought her life was going to get easier being around other supernaturals, but instead she meets a group of young werewolves that she seems to have rubbed the wrong way. The leader of that group is Derek; he's dark, tall, handsome, and a complete asshole. Yet, no matter how hard she tries to stay away, she just keeps getting pulled back to him. While she's trying to figure out her new life at a new school, someone else is planning on how to end it. Can she pull herself together long enough find this new threat? Or will he find her while she's looking the other way? The Prologue is posted please take a look and tell me what you think!!
Here is the book that started it all. Check it out and let me know what you think!!
Ash:In Training: Dominic always thought there was something wrong with her and on her sixteenth birthday she finds out what. She's not human; she a half-vampire, half-witch hybrid. And her real name is Ash. The people she always thought where her parents turn out to be the parents of dead child that she replaced. With threats coming in on all sides can Dom learn and accept who she really is fast enough? Or will she become dinner for those looking for her? Finished!! Take a look and let me know what you think!! PS It is the unedited version, so please don't comment on grammar or spelling.
The final book in the Ash trilogy (maybe) will be Ash: The Choice.
Ash: The Choice: Here is what I am thinking for the final book in the Ash trilogy. Five years later. Ash hasn't thought about Dominic, Joanna, or Terry Mason in a long time. After she moved from her childhood home to the bustling city, she left all that she once knew behind, as she thought that would be the best way to keep them safe. But someone else thinks it's the best way to stop Ash. Now Ash has to choose: Her parents or the ever growing group of supernaturals that depend on her to save them. Who will be her choice and will she be able to live with the consequences?
*Hiatus* The History: Human, Witch, Vampire, Shapeshifter: This will be a history book of sorts for Ash's world. It will contain the creation of the first four races and important wars, treaties, and events. Hope you check it out. Ran out of steam midway through the second chapter. Will try to go back to it eventually. Probably when I run into a rough patch with Ash.
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The Pakistan Government on Sunday wrote a letter to UN Secretary-General Ban Ki-moon pleading for assistance in border protection after at least 14 people were shot and killed near Kashmir. Pakistan has accused India of deliberately violating the established ceasefire by allowing cross border ...[read more]
An Algerian court on Monday sentenced former Guantanamo Bay detainee Abdul Aziz Naji to three years in prison for his participation in a foreign extremist group affiliated with al Qaeda. The sentence was less than the 10 years in prison and USD $6,330 fine sought by the prosecution. Naji was ...[read more]
Lawyers in Indian-administered Kashmir on Thursday called a one-day strike to demand the revocation of what they consider to be harsh emergency laws within their revolt-ridden region. The laws, the Armed Forces Special Powers Act (AFSPA) and the Disturbed Areas Act (DAA), were introduced in India ...[read more]
An anonymous official in India-controlled Kashmir reported Saturday that amendments to the Public Safety Act (PSA) have been approved. Prior to amendment, the law treated youth 16 years and older as adults, allowing them to be arrested. Now, no one under the age of 18 will be detained for ...[read more]
Kashmir's Chief Minister Omar Abdullah announced on Tuesday that officials will carry out DNA tests on the thousands of bodies recently uncovered in unmarked graves in the country's northern region. Abdullah called for families of missing persons to provide DNA samples for testing. The ...[read more]
Hundreds of people are being held without charge or trial in India's northernmost state Kashmir and Jammu, according to a report released by Amnesty International (AI) on Monday. AI reports that India's Public Safety Act (PSA) is being used to detain people despite the absence of sufficient ...[read more]
JURIST Contributing Editor Ali Khan of Washburn University School of Law says that the recent Mumbai massacre allegedly carried out by a militant organization fighting for liberation of Indian-occupied Kashmir is a sad reminder that the long-simmering dispute is the tinderbox of South Asia... ...[read more]
The Supreme Court of Pakistan on Friday ordered the release on bail of jailed opposition leader Javed Hashmi pending his appeal of his 2004 conviction for allegedly inciting mutiny in the armed forces, forgery and defamation by circulating an unsigned letter allegedly written by army officers ...[read more]
Indian authorities on Sunday arrested a fourth suspect in the July 11 Mumbai train bombings that killed over 180 people. Authorities arrested Dr. Tanyir Ansari late Sunday after several days of questioning in connection to the bombings. The Press Trust of India reports that Ansari is an alleged ...[read more]
Indian officials have named three suspects in Tuesday's Mumbai train bombings that killed some 180 people in seven different coordinated blasts after authorities detained 350 people for questioning Thursday. The suspects- Sayyad Zabiuddin and Zulfeqar Fayyaz named Thursday and a third man ...[read more]


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At the station, Alex is staring at the phone. She picks it up and puts it back down. "Who's that?" Jimmy asks. Alex says she's just making sure it's working. Lame. Alex is putting up that tough-girl front again with Jimmy. It's annoying as all-get-out. Jimmy suggests maybe she should ask if she can leave and go to the hospital. "I'm fine," she says, answering she'd just go crazy there, too. "Come on, Bobby," she says to the phone.
In the boxing ring, Bobby asks if Papa Was a Rolling Stone is there to apologize. "I'm not sorry," Papa says. "I had to be happy. I had to live." Bobby accuses him of abandoning them. "I just wanted a life of my own," Papa says. How dumb is that? Papa asks whether Bobby ever fought after that day. Flash to Young Bobby training with Papa.
Cut to Bosco and Yokas arriving at "Dollar Bill"'s house. It's the neighborhood where Paulie was looking for drugs earlier. "Get away from my door!" yells the guy inside. Yokas says they heard he had a visitor. "I'm gonna kill that little son of a bitch," Dollar Bill laughs. He says that Paulie is looking for the Greeks. Bosco gives the guy bad news: Narcotics plans to sweep his place in the morning. The guy says he keeps no drugs at home. But still. It sucks.
The next thing we see is Paulie approaching a bunch of druggies who look like extras from a Heart video. He asks everybody for drugs, and they shun him, like Heart would shun all those extras. They walk away and he gets nothing. Except maybe a worthless autograph from one of the Wilson sisters. He shuffles off.
Hospital. Kim is standing around when Gina bursts out of the elevator asking if Bobby's all right. "The hell are you doing here?" Kim asks her. "You don't belong here." Gina asks again about Bobby. Kim gives her a long look, gives another look to Ty and Sully, who are standing nearby, and walks off. Kim is in Silent But Deadly mode.
Doc and Carlos are giving blood. Doc says the boss said they can stay as long as they want to. Carlos says he needs to go get coffee. Huh?
In the hallway, a huge ass is skirting the top of the screen. Turns out it's Sully, approaching Kim, who is sitting alone. Sully sits next to her. It's Kim's big acting scene, but she doesn't say much. Sully says that Bobby always seemed like a nice guy who always helped people. Kim agrees. "He's a nice guy," she says, as if scorning him. "If he dies on that table, what did being a nice guy get him?" Probably a better show. D'oh, did I say that out loud? Kim walks away, probably to bitch at her agent for not ensuring she'd get more juicy scenes in this episode.


text: << Return to Meeting Archive Main Page SBE Work Session April 9, 2008 - 4/9/2008 9:00:00 AMColorado Department of Education201 E. Colfax Ave. Room 101Denver, CO 80203 Pamela Jo Suckla, Chairman, 3rd Congressional District Bob Schaffer, Vice-Chairman, 4th Congressional District Elaine Gantz Berman, 1st Congressional District Randy DeHoff, 6th Congressional District Evie Hudak, 2nd Congressional District Peggy Littleton, 5th Congressional District Jane Goff, 7th Congressional District Dwight Jones, Commissioner & Secretary to the State Board of Education Mary Frances Nevans, Director of State Board Relations Attendees - voting members Pamela Jo Suckla Chair, 3rd Congressional DistrictBob Schaffer Vice-Chair, 4th Congressional DistrictElaine Gantz Berman 1st Congressional District Randy G DeHoff 6th Congressional District Evie Hudak 2nd Congressional District Peggy Littleton 5th Congressional District 9:00 Welcome Chairwoman Suckla began the meeting with a statement responding to recent press that the SBE held a "secret" meeting or meetings in violation of the open meetings law (also known as the "Sunshine Law"). She explained that these meetings were executive sessions and that the meetings followed Colorado law permitting the SBE to confer with counsel in an ongoing lawsuit. The SBE's executive session on March 5 followed the court's denial of the SBE's request to be dismissed from the lawsuit. The SBE convened an executive session on March 5 to discuss with the attorneys what should be the next steps in litigation. Because of the high visibility of the lawsuit and the public interest in the SBE's decision about the ongoing suit, the statement read into the record during the March 6 meeting provided information about the Board's decision to defend the suit on the merits. Suckla said that it is not appropriate for Board members to disclose confidential information from an executive session. She said that although individual SBE members have chosen to express their views about the SBE's collective decision regarding this lawsuit, an individual Board member may not waive attorney-client privilege without the entire Board's approval. Chairman Suckla also mentioned that Board Member Jane Goff, who was appointed to the State Board on March 5, 2008, was out of the country on a trip scheduled prior to her appointment. SBE Reports 9:25 CDE Reports Communications Office Report SBE Office Report Presentation on Report of Financial Analysis by WestEd 10:15 Morning break 10:30 CDE Reports Recommended Changes to the SAR Because of bills passed last year, CDE is working on making changes to the School Accountability Reports (SARs). The most substantive change is in the calculation of the Academic Growth Rating, which will now be based on longitudinal growth according to the rules passed by the SBE last month. Many of the charts and tables on the SAR will change in terms of information presented. The panel "Questions Parents Should Ask" will be replaced with "School Information." The new SARs will be released this December. Board recommended changes to the Non-CSAP Course Offerings and Adequate Yearly Progress sections will be implemented by CDE staff. Recommendations for the Summer School Grant Program (1 CCR 301-50) 2008 Funded Sites. The State Board was asked to review and approve the proposed recommendations for the summer school grant recipients at the Regular Board Meeting on April 10, 2008. Recommendations to Summer School Funds for Eligible Facilities, Summer 2008 Summer School Grant Program for Eligible Facilities A grant program designed to assist facility schools in providing intensive research-based educational services in the areas of reading, writing, and/or mathematics for children who are performing below grade level in these areas. The summer school program is required to be at least six weeks long and at least 20 hours per week of academic instruction. This is a non-competitive grant. All below grade level students in eligible facilities that applied for the grant were approved for funding. The State Board was asked to approve a list of facilities and the recommended funding level for each at the April 10, 2008 Board meeting. 11:30 Lunch Executive Session A motion was made by Ms. Gantz Berman and seconded by Ms. Hudak to convene and executive session for the purpose of receiving legal advice from the Board's attorneys. 1:30 Rulemaking Hearing on the Permanent Rules for the Administration of the Accreditation of School Districts Ms. Littleton made a motion to close oral testimony at the conclusion of the public rulemaking hearing, direct recommended changes to the rules based on discussion and comment, and place the revised rules for Administration of the Accreditation of School Districts on the May 15 regular board meeting agenda for board discussion and adoption. The vote was seconded by Mr. DeHoff. Motion made by: Peggy Littleton Motion seconded by: Randy G DeHoff VotingUnanimously Approved 2:00 Legislation and Budget Legislation 2:30 Afternoon Break 3:00 CDE Reports 4:30 Issues and Trends Adjourn SecretaryChairperson


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$51-a-share offer by Swiss giant ups ante by 15 percent
"The bottom line is Illumina needs to keep the stock (price) up above the Roche offer," he said. "They're going to need to execute, get another bidder or whatever. But it doesn't look like there are multiple bidders here, and that is the key to getting a higher valuation."
In a letter to Illumina, Roche Chairman Franz Humer said the Swiss giant would prefer to negotiate a merger rather than continue its hostile takeover efforts.
"I strongly hope that you will either agree to commence discussions with us now or remove all obstacles so that your shareholders can make their own determinations about the adequacy of our increased offer," wrote Humer.
Illumina's shares traded up $2.52, or 5.05 percent, at $52.40 Thursday on the Nasdaq exchange.


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Tonight the Bellmont Braves host the Woodlan Warriors at the Teepee. The DDD will begin coverage at around 5:45 p.m. To view the gamecast simply click the link below then login with username and password for the service to be free of charge. Go Braves!
https://new.livestream.com/accounts/2267246/events/2963652 [2]
[1] http://www.decaturdailydemocrat.com/sites/default/files/1973b15d-790f-4a37-83f1-97f4d0c5c1cb_170x254.jpeg
[2] https://new.livestream.com/accounts/2267246/events/2963652
[3] http://www.decaturdailydemocrat.com/category/sports/local-sports


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Please join us for our second community meeting regarding the upcoming renovations to the play space at Guy Mason. Representatives from DPR, DGS and the design team will be on hand to present conceptual designs based on community input collected from prior meeting and online surveys. Staff members will also be on hand to answer any questions regarding the project.
Know before you go! Check out our Wait Times and Webcams.
Organ donation saves lives. If not already participating, sign up here.
Give us feedback! Engage with your government and help the DMV improve the quality of our services.


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Baby finally sleeping at night
Kim - 16th March 2015
I love this product!!! I had tried every type of swaddle and swaddle blanket for my baby. No matter what I tried, he would find a way work out of it and wake himself up, or scream because he doesn't like his legs contained. He needed his arms swaddled but...
No more crying in the car!!
kchinn - 10th April 2013
My third daughter is the only one who screams bloody murder in the car. I sat in the back with her for 2 hours trying everything to calm her down. She was just flailing her arms uncontrollably and getting more and more upset After pinning her arms down ...
Jill - 16th October 2012
Bought this for my 5th & final baby. Oh how I wish that I had purchased a woombie earlier! My last 3 babes have needed to be swaddled to sleep. We have used all types of swaddle blankets & wraps with limited success. The velcro wears out quickly & they wr...
Great for reflux babies
Glennen - 30th June 2012
My baby has reflux and sleeps on an inclined wedge with sling. The convertible leggies Woombie lets us swaddle her during bedtime routine instead of after she is placed in her sling. She loves it and goes right to sleep!
Works Wonders!
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My son has had severe eczema since he was one month old and will scratch until he bleeds. We used the miracle blanket until he started rolling around in his crib, then switched to the halo sleep sack swaddler, which work well until he learned to break fre...
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My son needs to be swaddled but he would break out of it every time he slept. We tried different products but he would still be able to break free from them. This is the only one that has helped him to sleep better. He has the freedom to move his arms b...


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Alien Skin Software makes creative tools for photographers and graphic designers. We distill advanced math and cutting edge research into simple tools that render beautiful pictures.
Recent Posts
Client Acquisition for Senior Portraits with Ari DorfmanHis unique approach to cultivating new client leads makes Ari Dorfman's senior portrait photography business stand out.
Photo of the Month
The next photo of the month award winner goes to Trish Hadley for her elegant portrait.


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I had regular gravel in one of my tanks and was looking for something to help plants thrive. I mixed this with the gravel and it did benefit the plants in the tank. They grew nicer and more quickly than they had been growing.
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P. Allen Smith | Grow Together, Go Together; Crudites | PBS
Garden and lifestyle expert P. Allen Smith shares and easy to prepare recipe to use at your next garden gathering! I love putting together finger foods when having friends over to the house. Their... 01:42
P. Allen Smith | Yummy Pepper Recipe : Roasted Baby Peppers and Cherry Tomatoes | PBS
Yummy baby bell peppers are bite-sized treats that you'll want to eat straight from the garden. They are nearly seedless and so sweet you'll find it hard to believe you are eating a pepper. They are... 01:38
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P. Allen Smith shows us how to make this festive and delicious fall recipe. 04:05
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Funny name, great dish. P. Allen Smith shows us how to make this easy and delicious recipe with vegetables from our gardens. 03:06
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Activities for Friends of the Memorial
Our special Friends-only activities give you an insider's view of the Memorial's exhibitions and galleries. The activities include behind-the-scenes tours and talks by the Memorial's historians and curators.
Bookings are essential for all Friends-only activities.
For further information or to make a booking to attend, please contact the Friends Coordinator on (02) 6243 4523 or email firstname.lastname@example.com
2015 Activity schedule
Victory in Europe (VE) Day Commemorative Art Tour: Bomber Crew
Thursday 7 May
9.00 am
Meet at the Administration Building
FREE, bookings essential
Stella Bowen's iconic painting Bomber Crew (ART26265) comes home! Bowen was one of only three women commissioned under Australia's official war art scheme during the Second World War.
Bomber Crew captures on canvas the experience, adventures and loss of Australian airmen who took part in the air war over Europe, and returns after touring with the Memorial's exhibition Reality in Flames: modern Australian art and the Second World War.
In conjunction with the 70th Anniversary of Victory in Europe (VE) Day, Friends of the Memorial are invited to join curators behind the scenes for an insight into Bowen's painting, the portrait sketches she made of the pilots on the day of their fateful mission, and the flying equipment they would have worn.
This special event takes place on Thursday 7 May and will culminate with a tour of a Lancaster bomber in Anzac Hall.
Please note this tour has limited places. Bookings are essential.
Bookings close Friday 1 May
Mother's Day White Glove Tour
Friday 8th May
10.00 am – 11.30 am
Education Space
FREE, Bookings Essential
Mothers and families of the Friends of the Memorial are invited to an exclusive "White Glove Tour" of the Memorial's collection to reflect on the strength and endurance of Australian mothers during times of war.
Meet with the curators behind the Memorial's world-class exhibitions this Mother's Day for a rare opportunity to handle objects from around the world carried by Australians at war.
Please note this tour has limited places. Bookings are essential.
Bookings close Thursday 7 May