Journal: Frontiers in Microbiology

Article Title: Fighting Ebola with novel spore decontamination technologies for the military

doi: 10.3389/fmicb.2015.00663

Figure Lengend Snippet: Spore strip bio-indicator characteristics.

Article Snippet: Three Bacillus spore species were assayed for susceptibility to ClO : B. atrophaeus (ATCC 9372, formerly B. subtilis var. niger and B. globigii ), B. thuringiensis (ATCC 29730), and G. stearothermophilus (ATCC 7953, formerly Bacillus stearothermophilus ).

Techniques: Stripping Membranes

Journal: Frontiers in Microbiology

Article Title: Fighting Ebola with novel spore decontamination technologies for the military

doi: 10.3389/fmicb.2015.00663

Figure Lengend Snippet: Tyvek spore strip data summary.

Article Snippet: Three Bacillus spore species were assayed for susceptibility to ClO : B. atrophaeus (ATCC 9372, formerly B. subtilis var. niger and B. globigii ), B. thuringiensis (ATCC 29730), and G. stearothermophilus (ATCC 7953, formerly Bacillus stearothermophilus ).

Techniques: Stripping Membranes

Journal: Frontiers in Microbiology

Article Title: Fighting Ebola with novel spore decontamination technologies for the military

doi: 10.3389/fmicb.2015.00663

Figure Lengend Snippet: (A) Statistical modeling showing the probability, with 95% confidence, of a single B. thuringiensis spore surviving a ClO 2 dose range of 438–4106 ppm-h at 79% RH. The spore population of each strip is fixed at 1.2 × 10 6 , and n = 98 each for strips contained in Tyvek or with no packaging. In the figure, the lighter set of dashed and solid lines represent the confidence interval for spore strips packaged in Tyvek or no packaging, respectively, while the middle bold lines represent the predicted probabilities. In this model, there is strong evidence of an effect of dose ( p = 0.002) and no evidence of an effect of packaging ( p = 0.76). (B) Statistical modeling showing the probability, with 95% confidence, of a single G. stearothermophilus spore surviving a ClO 2 dose range of 438–4106 ppm-h at 79% RH. The spore population of each strip is fixed at 1.0 × 10 6 , and n = 80 each for strips contained in Tyvek or no packaging. Lighter dashed and solid lines represent the confidence interval for spore strips packaged in Tyvek or no packaging, respectively, while the middle bold lines represent the predicted probabilities. There is evidence of an effect of dose ( p = 0.02) and no evidence of an effect of packaging ( p = 0.68). (C) Predicted probabilities of a single spore surviving for varying numbers of B. atrophaeus spores (1.2 × 10 4 to 1.2 × 10 7 ), with a D EtO value of 3.1, in Tyvek ( n = 152) and no packaging ( n = 144) at 79% RH and ClO 2 dose ranging from 110 to 1991 ppm-h. There is strong evidence that increased dose decreases the probability of any strips having live spores ( p < 0.01) and that as the number of spores on the strip increases, so does the probability of survival ( p < 0.01). There is no evidence of a significant effect of packaging on survival of spores ( p = 0.21), though there is some difference with a slightly higher rate of survival for spores packaged in Tyvek. (D) Predicted probabilities of a single spore surviving for varying numbers of B. atrophaeus spores (3.5 × 10 4 to 1.5 × 10 8 ), with a D EtO value of 5.0, in Tyvek ( n = 490) and no packaging ( n = 350) at 79% RH and ClO 2 dose ranging from 110 to 1991 ppm-h. These results indicate that the probability of a strip having live spores after treatment increases with decreasing dose ( p < 0.01) and with increasing numbers of spores ( p < 0.01). There is no evidence of an effect of packaging ( p = 0.21), although there is a slightly lower rate of survival for spores in Tyvek packagi.

Article Snippet: Three Bacillus spore species were assayed for susceptibility to ClO : B. atrophaeus (ATCC 9372, formerly B. subtilis var. niger and B. globigii ), B. thuringiensis (ATCC 29730), and G. stearothermophilus (ATCC 7953, formerly Bacillus stearothermophilus ).

Techniques: Stripping Membranes

Journal: Frontiers in Microbiology

Article Title: Fighting Ebola with novel spore decontamination technologies for the military

doi: 10.3389/fmicb.2015.00663

Figure Lengend Snippet: Increasing RH at a fixed dose of (A) 500 ppm ClO 2 for 4 h (∼2000 ppm-h) and (B) 1000 ppm ClO 2 for 4 h (∼4000 ppm-h) results in a higher percentage of log 6 B. atrophaeus spore strips being killed. (A) B. atrophaeus D EtO = 3.3 ( n = 120), = B. atrophaeus D EtO = 3.8 ( n = 160); (B) B. atrophaeus D EtO = 3.3 ( n = 140), and B. atrophaeus D EtO = 3.8 ( n = 140). Data shown is for spore strips packaged in Tyvek. For the D EtO = 3.3 strips exposed to ∼2000 ppm-h ClO 2 , ≥80% kill is achieved at 70 %RH and above, but when we increase the dose to ∼4000 ppm-h the same level of kill is achieved at %RH of 60. The relationship between low D EtO value and increased susceptibility to ClO 2 is evident, though, in this example, we see that at 90 %RH all four sample sets are behaving roughly the same. (C) B. atrophaeus D EtO = 3.3, log 6 spore strips packaged in img medical-grade glassine, and img Tyvek are not equally susceptible to a set ClO 2 dose at 79 %RH (NE = no effect). Spore strips were exposed to 250 ppm ClO 2 for 4 h (dose ≈1000 ppm-h), where n = 20 each for glassine and Tyvek; 2000 ppm-h, where n = 40 (glassine) and n = 20 (Tyvek); and 4000 ppm-h, where n = 40 (glassine) and n = 20 (Tyvek) at constant temperature and %RH. (D) We did not achieve 100% kill for glassine-packaged log 6 spore strips until we subjected a lot of B. atrophaeus with D EtO value of 3.0 ( n = 20) to 2000 ppm for 10 h (∼20,000 ppm-h) at 79 %RH. This high dose killed 80 and 85% of B. atrophaeus D EtO = 3.3 and 3.8 ( n = 20 each) spore strips, respectively. (E) For all three species and B. atrophaeus with two different D EtO values (3.3, with n = 80 for each Tyvek and no packaging; and 3.8, with n = 189 for each Tyvek and no packaging), probabilities were predicted for a spore surviving after treatment, with RH = 0.79 and N = 10 6 . Note that for B. thuringiensis ( n = 98 each for Tyvek and no packaging), measurements were only made at N = 1.2 × 10 6 . For both sets of B. atrophaeus , measurements were made at N = 1.3 × 10 6 , so in these cases this plot involves extrapolation in N . B. thuringiensis , the closest phylogenetic relative to Bacillus anthracis used in this study, is more likely to survive at any given ClO 2 dose than either of the B. atrophaeus strains, though it’s resistance is similar to the strain of G. stearothermophilus ( n = 80 each for Tyvek and no packaging) assayed.

Article Snippet: Three Bacillus spore species were assayed for susceptibility to ClO : B. atrophaeus (ATCC 9372, formerly B. subtilis var. niger and B. globigii ), B. thuringiensis (ATCC 29730), and G. stearothermophilus (ATCC 7953, formerly Bacillus stearothermophilus ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Fighting Ebola with novel spore decontamination technologies for the military

doi: 10.3389/fmicb.2015.00663

Figure Lengend Snippet: Spore height determinations.

Article Snippet: Three Bacillus spore species were assayed for susceptibility to ClO : B. atrophaeus (ATCC 9372, formerly B. subtilis var. niger and B. globigii ), B. thuringiensis (ATCC 29730), and G. stearothermophilus (ATCC 7953, formerly Bacillus stearothermophilus ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Fighting Ebola with novel spore decontamination technologies for the military

doi: 10.3389/fmicb.2015.00663

Figure Lengend Snippet: Characterization of Bacillus spores. (A,B) AFM images of native air-dried spores. (A) Height image of B. anthracis Sterne spores and (B) amplitude image of B. thuringiensis spores. In both images surface ridges extending along the entire length of spores (several surface ridges noted by white arrows) are seen. In (A) a collapsed spore is indicated with a black arrow. In (B) an exosporia is indicated with Ex. (C,D) Phase contrast microscopy images of B. anthracis Sterne spores (C) and B. thuringiensis spores (D) .

Article Snippet: Three Bacillus spore species were assayed for susceptibility to ClO : B. atrophaeus (ATCC 9372, formerly B. subtilis var. niger and B. globigii ), B. thuringiensis (ATCC 29730), and G. stearothermophilus (ATCC 7953, formerly Bacillus stearothermophilus ).

Techniques: Microscopy

Journal: Frontiers in Microbiology

Article Title: Fighting Ebola with novel spore decontamination technologies for the military

doi: 10.3389/fmicb.2015.00663

Figure Lengend Snippet: AFM images of γ-irradiated (≥2.5 × 10 6 rads) B. thuringiensis spores in (A) a low magnification AFM image. In a higher magnification AFM image (B) , exosporia (denoted with Ex) are observed. Image (C) is a close-up image taken of the lower right area in panel (B) .

Article Snippet: Three Bacillus spore species were assayed for susceptibility to ClO : B. atrophaeus (ATCC 9372, formerly B. subtilis var. niger and B. globigii ), B. thuringiensis (ATCC 29730), and G. stearothermophilus (ATCC 7953, formerly Bacillus stearothermophilus ).

Techniques: Irradiation

Journal: Experimental & Molecular Medicine

Article Title: Increased arginase II activity contributes to endothelial dysfunction through endothelial nitric oxide synthase uncoupling in aged mice

doi: 10.3858/emm.2012.44.10.068

Figure Lengend Snippet: Upregulation of ArgII activity in aged mice aortas. (A) Western blot analysis of aortas from young and aged mice. Similar levels of ArgII were found in both groups, but ArgI protein was not detectable (upper). The lower panel shows the densitometric analysis, n = 4 experiments. (B) ArgI and ArgII mRNA expression levels were analyzed by RT-PCR using GAPDH as a control. Only ArgII mRNA was detected (upper). The lower panel shows the densitometric analysis, n = 4 experiments. (C) siArgII incubation decreased arginase activity in both young and aged aortas ( # vs. young, P < 0.05; * vs. young, P < 0.01; ** vs. aged, P < 0.01). (D) siArgII effectively downregulated ArgII protein expression in both young and old aortas ( # vs. young, P < 0.05; * vs. aged, P < 0.01), but not with siArgI ( ## vs. aged, ns).

Article Snippet: Thoracic aortas were dissected from euthanized mice, cut into 1.5-mm rings, and incubated overnight in Dulbecco's modified Eagle's medium (DMEM) containing siArgI and/or siArgII (100 nM each, Santa Cruz Biotechnology, sc-29727 and sc-29730, respectively).

Techniques: Activity Assay, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Incubation

Journal: Experimental & Molecular Medicine

Article Title: Increased arginase II activity contributes to endothelial dysfunction through endothelial nitric oxide synthase uncoupling in aged mice

doi: 10.3858/emm.2012.44.10.068

Figure Lengend Snippet: Increased ArgII activity was associated with eNOS uncoupling. (A) Increased eNOS expression in aged aorta was not attenuated by siArgII ( * vs. young, P < 0.01; ** vs. aged, ns). (B) siArgII incubation induced eNOS coupling in young aorta ( # vs. young, P < 0.01). Augmented eNOS uncoupling in aged aorta was decreased by siArgII. Incubation of aged aorta with the arginase inhibitor, ABH (10 µM), enhanced eNOS dimerization ( * vs. young, P < 0.01; ** vs. aged, P < 0.01). Densitometric analysis of Western blot (lower panel).

Article Snippet: Thoracic aortas were dissected from euthanized mice, cut into 1.5-mm rings, and incubated overnight in Dulbecco's modified Eagle's medium (DMEM) containing siArgI and/or siArgII (100 nM each, Santa Cruz Biotechnology, sc-29727 and sc-29730, respectively).

Techniques: Activity Assay, Expressing, Incubation, Western Blot

Journal: Experimental & Molecular Medicine

Article Title: Increased arginase II activity contributes to endothelial dysfunction through endothelial nitric oxide synthase uncoupling in aged mice

doi: 10.3858/emm.2012.44.10.068

Figure Lengend Snippet: Inhibition of ArgII activity restored impaired endothelial function in aged mice. Isolated aortic segments were incubated with DAF-AM (5 µM) and fluorescence was measured in real-time (endothelium side up). The slope of DAF fluorescence was determined. (A) siArgII incubation with young vessels increased the slope of DAF fluorescence ( # vs. young, P < 0.01). However, DAF fluorescence was decreased in aged mouse aorta and increased following siArgII (but not siArgI) and ABH incubation. L-NAME was used as a control ( * vs. young, P < 0.01; ** vs. aged, P < 0.01; n = 4 mice). (B) ROS production in aortic endothelium was measured with DHE (5 µM), and the slope of DHE fluorescence was determined using cumulative data. Arginase inhibition with siArgII and ABH reciprocally regulated ROS production ( # vs. young, P < 0.01; * vs. young, P < 0.01; ** vs. aged, P < 0.01; ## vs. aged, P < 0.01; n = 4 mice). MnTBAP was used as a control.

Article Snippet: Thoracic aortas were dissected from euthanized mice, cut into 1.5-mm rings, and incubated overnight in Dulbecco's modified Eagle's medium (DMEM) containing siArgI and/or siArgII (100 nM each, Santa Cruz Biotechnology, sc-29727 and sc-29730, respectively).

Techniques: Inhibition, Activity Assay, Isolation, Incubation, Fluorescence, Control

Journal: Experimental & Molecular Medicine

Article Title: Increased arginase II activity contributes to endothelial dysfunction through endothelial nitric oxide synthase uncoupling in aged mice

doi: 10.3858/emm.2012.44.10.068

Figure Lengend Snippet: siArgII improved impaired vascular reactivity in aged mice. (A) Aged aortic vessels had augmented contractile responses to U46619 compared to young mice ( * , young vs. aged, P < 0.01). siArgII restored the U46619-mediated pressor response of aged aortic vessels to levels similar to that of young aortas ( * , young vs. aged, P < 0.01; ** , aged vs. aged + siArgII, P < 0.01). n = 4 mice per each group. (B) siArgII attenuated contractile responses to U46619 in young aorta ( * , young vs. young + siArgII, P < 0.01) and ABH incubation with old aorta significantly retarded U46619-dependent constriction ( ** , aged vs. aged + ABH, P < 0.01). (C) Endothelium-dependent relaxation responses to Ach were impaired in aged aortas ( * , young vs. aged, P < 0.01). Impaired relaxation was recovered with siArgII pretreatment ( ** , aged vs. aged + siArgII, P < 0.05). n = 4 mice per each group. (D) ABH incubation augmented Ach-dependent vessel relaxation in aged aorta ( * , aged vs. aged + ABH, P < 0.01).

Article Snippet: Thoracic aortas were dissected from euthanized mice, cut into 1.5-mm rings, and incubated overnight in Dulbecco's modified Eagle's medium (DMEM) containing siArgI and/or siArgII (100 nM each, Santa Cruz Biotechnology, sc-29727 and sc-29730, respectively).

Techniques: Incubation

Journal: Experimental & Molecular Medicine

Article Title: Increased arginase II activity contributes to endothelial dysfunction through endothelial nitric oxide synthase uncoupling in aged mice

doi: 10.3858/emm.2012.44.10.068

Figure Lengend Snippet: siArgII preincubation increased intracellular L -arg concentrations. Aortic vessels were incubated with or without siArgI, siArgII, and ABH and intracellular L -arg concentrations were measured with HPLC (see methods section). Treatment of siArgII to young vessels resulted in increase of L -arg amount ( # vs. young, P < 0.01). The reduced L -arg level in aortas from aged mice was restored to that of young aortas by incubation of siArgII and ABH ( * vs. young, P < 0.01; ** vs. aged, P < 0.01; n = 6).

Article Snippet: Thoracic aortas were dissected from euthanized mice, cut into 1.5-mm rings, and incubated overnight in Dulbecco's modified Eagle's medium (DMEM) containing siArgI and/or siArgII (100 nM each, Santa Cruz Biotechnology, sc-29727 and sc-29730, respectively).

Techniques: Incubation