Environmental persistence of vaccinia virus on materials.

Smallpox is caused by the variola virus, and ranks as one of the most serious diseases that could originate from a biological weapon. However, limited data exist on the persistence of variola and related viruses on materials (that may act as fomites), under controlled environmental conditions. To fill these data gaps, we determined the persistence of the vaccinia virus (an established surrogate for the variola virus) as a function of temperature, relative humidity and material. Experiments were conducted with vaccinia virus in a freeze-dried form, using four materials under four sets of environmental conditions. After elapsed times ranging from 1 to 56 days, the virus was extracted from small coupons and quantified via plaque-forming units (PFU). The vaccinia virus was most persistent at low temperature and low relative humidity, with greater than 10(4) PFU recovered from glass, galvanized steel and painted cinder block at 56 days (equivalent to only a c. 2 log reduction). Thus, vaccinia virus may persist from weeks to months, depending on the material and environmental conditions. This study may aid those responsible for infection control to make informed decisions regarding the need for environmental decontamination following the release of an agent such as variola.

Efficacy of liquid spray decontaminants for inactivation of Bacillus anthracis spores on building and outdoor materials.

AIMS: To obtain data on the efficacy of various liquid and foam decontamination technologies to inactivate Bacillus anthracis Ames and Bacillus subtilis spores on building and outdoor materials. METHODS AND RESULTS: Spores were inoculated onto test coupons and positive control coupons of nine different materials. Six different sporicidal liquids were spray-applied to the test coupons and remained in contact for exposure times ranging from 10 to 70 min. Following decontamination, spores were recovered from the coupons and efficacy was quantified in terms of log reduction. CONCLUSIONS: The hydrogen peroxide/peracetic acid products were the most effective, followed by decontaminants utilizing hypochlorous acid chemistry. Decontamination efficacy varied by material type. SIGNIFICANCE AND IMPACT OF THE STUDY: The study results may be useful in the selection of technologies to decontaminate buildings and outdoor areas in the event of contamination with B. anthracis spores. These results may also facilitate selection of decontaminant liquids for the inactivation of other spore-forming infectious disease agents.

Decreased time for detection and quantification of virulent Bacillus anthracis and Yersinia pestis using a BioNanoPore (BNP) membrane technology.

Many aspects of biodefense research require quantitative growth assessments of the test agent. This study evaluated the BioNanoPore (BNP) technology to quantitate Bacillus anthracis and Yersinia pestis faster than traditional plate counting methods. The BNP technology enabled quantification of B. anthracis and Y. pestis in phosphate-buffered saline and naïve rabbit blood at 6 and 24 h, respectively. After 6 h of growth, counts for B. anthracis ranged from 6.19-6.45 log(10) CFU ml(-1) on BNP, while counts after 24 h on tryptic soy agar (TSA) ranged from 6.51-6.58 log(10) CFU ml(-1). For Y. pestis, counts on BNP at 24 h ranged from 6.31-6.41 log(10) CFU ml(-1) on BNP and ranged from 6.44-6.89 log(10) CFU ml(-1) on TSA at 48 h. This study demonstrates that the BNP technology provides a more rapid detection of B. anthracis and Y. pestis, which could aid in the evaluation of potential medical countermeasures and treatments as well as other biological defense applications such as surface sampling or decontamination efficacy.

Use of superabsorbent polymer gels for surface decontamination of Bacillus anthracis spores.

AIMS: This study evaluated the inactivation of Bacillus anthracis Vollum spores dried on a nonporous surface using a superabsorbent polymer (SAP) gel containing commercially available liquid decontaminants. METHODS AND RESULTS: The first phase determining the availability of the liquid decontaminant within the SAP showed that the SAP gel containing pH-adjusted sodium hypochlorite (NaOCl) inhibited B. anthracis growth while the water control SAP gel had no affect on growth. For testing surface decontamination, B. anthracis spores were dried onto steel coupons painted with chemical agent resistant coating and exposed to SAP containing either pH-adjusted NaOCl, chlorine dioxide (ClO(2)) or hydrogen peroxide/peracetic acid (H(2)O(2)/PA) for 5 and 30 min. At contact times of both 5 and 30 min, all of the SAP gels containing pH-adjusted NaOCl, ClO(2) or H(2)O(2)/PA inactivated B. anthracis spores at levels ranging from 2.2 to > or =7.6 log reductions. CONCLUSIONS: Incorporation of three commercially available decontaminant technologies into a SAP gel promotes inactivation of B. anthracis spores without observable physical damage to the test surface. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides preliminary data for the feasibility of using SAP in inactivating B. anthracis spores on a nonporous surface, supporting the potential use of SAP in surface decontamination.

Vapour-phase hydrogen peroxide inactivates Yersinia pestis dried on polymers, steel, and glass surfaces.

AIMS: This study evaluated the inactivation of virulent Yersinia pestis dried on polymers, steel, and glass surfaces using vapour-phase hydrogen peroxide. METHODS AND RESULTS: A suspension of Y. pestis CO92 (1.70 x 10(8) CFU) was dried on 10 different types of test surfaces and exposed to vapour-phase hydrogen peroxide fumigation for a contact time of 2 h. A significant reduction in the log10 CFU of Y. pestis on all 10 materials was observed between the controls evaluated after a 1 h drying time and unexposed controls evaluated after the decontamination run. Qualitative growth assessment showed that vapour-phase hydrogen peroxide exposure inactivated Y. pestis on all replicates of the 10 test materials as well as biological indicators up to 7 days postexposure. CONCLUSIONS: Virulent Y. pestis CO92 is inactivated on polymers, steel, and glass surfaces when exposed to vapour-phase hydrogen peroxide without observable physical damage to the test materials. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information for using vapour-phase hydrogen peroxide as a practical process for the decontamination of virulent Y. pestis in circumstances where time-dependent attenuation/inactivation orliquid/heat decontamination may not be the most suitable approach.

Formaldehyde gas inactivation of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface materials.

AIMS: To evaluate the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface materials using formaldehyde gas. METHODS AND RESULTS: B. anthracis, B. subtilis, and G. stearothermophilus spores were dried on seven types of indoor surfaces and exposed to approx. 1100 ppm formaldehyde gas for 10 h. Formaldehyde exposure significantly decreased viable B. anthracis, B. subtilis, and G. stearothermophilus spores on all test materials. Significant differences were observed when comparing the reduction in viable spores of B. anthracis with B. subtilis (galvanized metal and painted wallboard paper) and G. stearothermophilus (industrial carpet and painted wallboard paper). Formaldehyde gas inactivated>or=50% of the biological indicators and spore strips (approx. 1x10(6) CFU) when analyzed after 1 and 7 days. CONCLUSIONS: Formaldehyde gas significantly reduced the number of viable spores on both porous and nonporous materials in which the two surrogates exhibited similar log reductions to that of B. anthracis on most test materials. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide new comparative information for the decontamination of B. anthracis spores with surrogates on indoor surfaces using formaldehyde gas.

A preliminary assessment of Bacillus anthracis spore inactivation using an electrochemically activated solution (ECASOL).

AIM: To evaluate the efficacy of electrochemically activated solution (ECASOL) in decontaminating Bacillus anthracis Ames and Vollum 1B spores, with and without changing the source water hardness and final ECASOL pH. METHODS AND RESULTS: Five different ECASOL formulations were generated, in which the source water hardness and final ECASOL pH were varied, resulting in cases where significant changes in free available chlorine (FAC) and oxidative-reduction potential (ORP) were observed. B. anthracis Ames and Vollum 1B spores were suspended in the various ECASOL formulations for 30 min, and decontamination efficacy was determined; calcium hypochlorite [5% high-test hypochlorite (HTH)] was used as a positive control. The five different ECASOL formulations yielded mean FAC levels ranging from 305 to 464 ppm, and mean ORP levels ranging from +826 to +1000 mV. Exposure to all the ECASOL formulations and 5% HTH resulted in >or=7.0 log reductions in both B. anthracis Ames and Vollum 1B spores. CONCLUSIONS: The present testing demonstrated that ECASOL with a minimum of c. 300-ppm FAC levels and +800-mV ORP inactivated the B. anthracis spores in suspension, similar to 5% HTH. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide information for decontaminating B. anthracis Ames and Vollum 1B spores in suspension using ECASOL.

Decontamination assessment of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surfaces using a hydrogen peroxide gas generator.

AIMS: To evaluate the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface materials using hydrogen peroxide gas. METHODS AND RESULTS: Bacillus anthracis, B. subtilis, and G. stearothermophilus spores were dried on seven types of indoor surfaces and exposed to > or =1000 ppm hydrogen peroxide gas for 20 min. Hydrogen peroxide exposure significantly decreased viable B. anthracis, B. subtilis, and G. stearothermophilus spores on all test materials except G. stearothermophilus on industrial carpet. Significant differences were observed when comparing the reduction in viable spores of B. anthracis with both surrogates. The effectiveness of gaseous hydrogen peroxide on the growth of biological indicators and spore strips was evaluated in parallel as a qualitative assessment of decontamination. At 1 and 7 days postexposure, decontaminated biological indicators and spore strips exhibited no growth, while the nondecontaminated samples displayed growth. CONCLUSIONS: Significant differences in decontamination efficacy of hydrogen peroxide gas on porous and nonporous surfaces were observed when comparing the mean log reduction in B. anthracis spores with B. subtilis and G. stearothermophilus spores. SIGNIFICANCE AND IMPACT OF THE STUDY: These results provide comparative information for the decontamination of B. anthracis spores with surrogates on indoor surfaces using hydrogen peroxide gas.

Detection of oxidative species and low-molecular-weight DNA in skin following dermal exposure with JP-8 jet fuel.

Dermal absorption of JP-8 jet fuel can lead to skin irritation within hours after exposure. This study detected the formation of oxidative species and low-molecular-weight DNA in rat skin as potential indicators of JP-8-induced skin injury. At 0, 1, 2, 4 and 6 h after the beginning of a 1-h exposure, skin samples were removed and analyzed for oxidative species formation and low-molecular-weight DNA analysis. At 1, 2 and 4 h, mean oxidative species levels increased significantly (P < 0.05) above unexposed samples. Significantly higher (P < 0.05) low-molecular-weight DNA values were observed at 4 and 6 h compared with unexposed controls. These results demonstrate significant increases in oxidative species and low-molecular-weight DNA levels in the skin following dermal exposure to JP-8. These responses may serve as indicators of skin injury following exposure to JP-8 jet fuel and other volatile chemicals or mixtures.

Dermal exposure to m-xylene leads to increasing oxidative species and low molecular weight DNA levels in rat skin.

Dermal absorption of organic solvents, such as m-xylene, can lead to skin inflammation and pathological changes within hours after exposure. This study detected oxidative species formation and low molecular weight (LMW) DNA in rat skin as potential indicators of m-xylene-induced skin injury. At 0, 1, 2, 4, and 6 h after the beginning of a 1-h exposure, skin samples were removed and analyzed for oxidative species formation and LMW DNA analysis. At 2 h, mean oxidative species levels increased significantly (P < 0.05) above unexposed samples. Significantly higher (P < 0.05) LMW DNA values were observed at 2, 4, and 6 h compared to unexposed controls. These results show that oxidative species formation and LMW DNA levels in the skin may serve as indicators for predicting safe exposure levels to m-xylene and other volatile organic solvents.

Effect of JP-8 jet fuel on molecular and histological parameters related to acute skin irritation.

Organic chemicals such as jet fuels and solvents can cause skin irritation after dermal exposure. The molecular responses to these chemicals resulting in acute irritation are not understood well enough to establish safe exposure limits. Male F-344 rats were dermally exposed to JP-8 jet fuel for 1 h using Hill Top Chambers. Whole skin samples were collected at 0, 1, 2, 4, and 6 h after the beginning of the exposures, homogenized, and analyzed for interleukin (IL)-1alpha and inducible nitric oxide synthase (iNOS) protein and nitrite levels. IL-1alpha levels (determined by ELISA) ranged from approximately 11 to 34% above the 0-h samples over the observed time period. At 1 and 2 h, significantly higher (p < 0.05) levels of IL-1alpha were detected when compared to the 0-h samples. Western blot analysis revealed significantly higher (p < 0.05) levels of iNOS at 4 and 6 h compared to 0-h samples. Increases in IL-1alpha and iNOS expression were also observed in the skin immunohistochemically. Nitrite concentrations in skin samples were measured to estimate nitric oxide production. Although nitrite concentrations in the skin increased approximately 6-27% above the 0-h samples over the observed time period, no significant changes in nitrite levels were detected. Pathological changes in the skin following JP-8 exposure were evaluated histologically. Increased numbers of granulocytes were observed infiltrating the skin at 2 h and were more prominent by 6 h. These data show that a 1-h exposure to JP-8 results in a local inflammatory response, which can be detected by changes in molecular and histological parameters.

The relationship between chronic ankle instability and variations in mortise anatomy and impingement spurs.

Thirty-five patients undergoing a Bröstrom procedure for ankle instability were studied retrospectively as to the presence or absence of spurs and loose bodies, outcome, and mortise relationships. 100 adult volunteers had their ankles radiographically and clinically examined for spurs, loose bodies, and laxity. 100 patients' ankles with computed axial tomography were examined to define malleolar relationships. The AOFAS Hindfoot scores on the Bröstrom patients with or without spurs were not different. Patients undergoing a Bröstrom procedure had a 3.37 times incidence of spurs and/or loose bodies compared to normal adult population. The incidence of asymmetric but asymptomatic ankle laxity in normal adults was 11%. The fibula has a 38 degree range of position relative to the axis of the talus and the medial malleolus. A posterior fibular position may predispose to injury.

Murine response to DNA encoding herpes simplex virus type-1 glycoprotein D targeted to the liver.

Plasmid DNA encoding herpes simplex virus type-1 glycoprotein D (gD-1) was complexed with asialoorosomucoid conjugated to poly-L-lysine. Following its intravenous injection into BALB/c mice, this complex was targeted to the liver. Liver cells expressing gD-1 were detected immunohistochemically through day 6 post-immunization, while gD-1 DNA was detectable through 14 days post-immunization. Decline of gD-1 expression and detectable gD-1 DNA in the liver correlated with influx of T cells, predominantly CD4(+). The ASOR-poly-L-lysine DNA carrier system promotes hepatic expression of gD-1 and may be useful in vaccination against herpes simplex virus type-1.

Targeted delivery of DNA encoding herpes simplex virus type-1 glycoprotein D enhances the cellular response to primary viral challenge.

Intravenous injection of plasmid DNA encoding herpes simplex virus type-1 glycoprotein D (gD-1) complexed with asialoorosomucoid-poly-L-lysine (gD-ASOR) targets foreign DNA to the liver, leading to hepatic expression of gD-1. BALB/c mice were given two intravenous injections of gD-ASOR, pBK-ASOR (plasmid lacking the gD-1 gene but complexed with ASOR), or PBS. The skin was inoculated with 1 x 10(4) PFU of HSV-1 or sham-inoculated, and analyzed for infectious virus and cellular infiltration 1, 3, and 5 days after inoculation. Prior immunization with gD-ASOR led to significantly lower (P < 0.05) viral titers in the skin 5 days after inoculation compared with controls. Infiltration of the skin at the site of inoculation by polymorphonuclear neutrophils (PMNs), T cells, B cells, dendritic cells, and macrophages was monitored immunohistochemically. Significantly higher numbers (P < 0.05) of CD4+ and CD8+ T cells, dendritic cells, and macrophages responded to HSV-1 challenge in mice immunized with gD-ASOR than in mice immunized with pBK-ASOR or PBS. The response by PMNs and B cells was indistinguishable among the treatment groups. These results suggest that BALB/c mice sensitized to gD-1 following gD-ASOR immunization develop an enhanced T-cell response to primary HSV-1 infection.

Differential response of basal keratinocytes in a human skin equivalent to ultraviolet irradiation.

The human skin equivalent (HSE) provides a convenient model system for studying the cellular responses of basal keratinocytes to UV irradiation. HSEs, constructed by overlaying a collagen-fibroblast matrix with epidermal cells, were raised to an air-liquid interface to promote epidermal differentiation. HSEs were exposed to ultraviolet radiation from a 500-W Hot Quartz Hanovia therapeutic sunlamp, at a total dose of 100 J/m2. The HSEs were then frozen every 4 h over a 48-h period and cryosectioned. For each time period, the expression of beta1 integrin and cyclin E, p53, or Bcl-2 were quantified using dual immunolocalization. Basal cells expressing beta1 integrin were divided into two subpopulations, denoted beta1high or beta1low. The proportion of beta1high keratinocytes expressing Bcl-2 and cyclin E increased significantly 4 and 8 h, respectively, after exposure to UV; during the subsequent 16 h, this basal cell subpopulation expressed p53. By contrast, significant numbers of beta1low basal keratinocytes expressed p53, but not Bcl-2. These results suggest that beta1high and beta1low populations of basal epidermal cells in HSEs respond differently to UV irradiation.

Thoracolumbar burst fractures: CT dimensions of the spinal canal relative to postsurgical improvement.

Cross-sectional spinal canal area was measured before and after surgery in 12 patients with thoracolumbar burst fractures and canal narrowing caused by retropulsed fragments. Patients were classified into Denis type A or type B. Denis type A fractures have comminution of both end-plates of the vertebral body creating multiple smaller fractures; Denis type B fractures have comminution of the superior end-plate only with a single vertical fracture line into the inferior end-plate creating larger fragments. The degree of neurologic impairment was assessed before and after surgery using the Frankel system. There was no correlation between degree of canal narrowing and degree of neurologic impairment. The degree of spinal canal narrowing reflects the final resting position of the vertebral body fragments after trauma; during trauma, greater degrees of canal impingement may have occurred. Also, significant canal narrowing may be present without pinching of the cord or cauda equina. All patients with Denis type A fractures had near-anatomic reduction of fragments out of the spinal canal by surgery; less than half of the patients with Denis type B had good reduction. There was no correlation between reduction of retropulsed fragments and subsequent neurologic improvement. However, this should not preclude surgery as a therapeutic option: Eight of 10 patients with neurologic impairment experienced some improvement in symptoms after surgery; the other two were unchanged.

Intraoperative neurosonography: application and technique.

Intraoperative neurosonography was performed in 44 patients with contact transdural or transgyral scanning technique. Localization of intracranial pathology included primary brain tumors (24), metastatic tumors (11), aneurysms (two), abscesses (two), arteriovenous malformation (one), thrombosed arteriovenous malformations (two), and plasmacytoma (one). Sonographic guidance was used in transdural decompression of three cystic lesions, therapeutic and diagnostic aspiration of two abscesses, and biopsy of three solid lesions. The expertise of the physician-sonographer with sonographic equipment facilitates accurate and expedient intraoperative neurosurgical localization of pathology.

Low sensitivity of sonography and cholescintigraphy in acalculous cholecystitis.

Acalculous cholecystitis is difficult to diagnose by clinical means or contrast radiography. Because sonography and cholescintigraphy have both been shown to do well in the diagnosis of calculous cholecystitis, the sensitivity of these newer imaging methods was assessed retrospectively in 33 proven cases of acalculous cholecystitis. The sensitivities to acalculous cholecystitis for sonography (67%) and for cholescintigraphy (68%) were not as high as has been reported for these tests in calculous cholecystitis. Reasons for the lower sensitivity with each test and the pathogenesis of acalculous cholecystitis are discussed.