Combining spore germination and heat inactivation to decontaminate materials contaminated with Bacillus anthracis spores.

AIMS: To add a spore germination step in order to reduce decontamination temperature and time requirements compared to the current hot, humid air decontamination parameters, which are 75-80°C, ≥72 h, 70-90% RH, down to ≤60°C and ≤24 h total decontamination time. METHODS AND RESULTS: Bacillus anthracis spore germination with l-alanine+inosine+calcium dipicolinate (CaDPA) was quantified at 0-40°C, several time points and spore concentrations of 5-9 log10 per ml. Germination efficiency at 0-40°C was >99% at <8 log10 spores per ml. The temperature optimum was 20°C. Germination efficiency was significantly higher but slower at 0°C compared to ≥30°C at ≥8 log10 spores per ml. A single germinant application followed by 60°C, 1-h treatment consistently inactivated >2 log10 (>99%) of spores. However, a repeat application of germinant was needed to achieve the objective of ≥6 log10 spore inactivation out of a 7 log10 challenge (≥99·9999%) for ≤24 h total decontamination time for nylon and aircraft performance coating. CONCLUSIONS: l-alanine+inosine+CaDPA stimulated germination across wide temperature and spore concentration ranges. SIGNIFICANCE AND IMPACT OF THE STUDY: Germination expands the scope of spore decontamination to include materials from any industry sector that can be sprayed with an aqueous germinant solution.

Probing the structures of gas-phase rhodium cluster cations by far-infrared spectroscopy.

The geometric structures of small cationic rhodium clusters Rh(n)(+) (n = 6-12) are investigated by comparison of experimental far-infrared multiple photon dissociation spectra with spectra calculated using density functional theory. The clusters are found to favor structures based on octahedral and tetrahedral motifs for most of the sizes considered, in contrast to previous theoretical predictions that rhodium clusters should favor cubic motifs. Our findings highlight the need for further development of theoretical and computational methods to treat these high-spin transition metal clusters.

Communications: The structure of Rh(8) (+) in the gas phase.

The geometric structure of the Rh(8) (+) cation is investigated using a combination of far-infrared multiple photon dissociation spectroscopy and density functional theory (DFT) calculations. The energetic ordering of the different structural motifs is found to depend sensitively on the choice of pure or hybrid exchange functionals. Comparison of experimental and calculated spectra suggests the cluster to have a close-packed, bicapped octahedral structure, in contrast to recent predictions of a cubic structure for the neutral cluster. Our findings demonstrate the importance of including some exact exchange contributions in the DFT calculations, via hybrid functionals, when applied to rhodium clusters, and cast doubt on the application of pure functionals for late transition metal clusters in general.

Gastric cancer: establishing predictors of biologic behavior with use of population-based data.

BACKGROUND: Tumor thickness and nodal status are important predictors of survival following curative resection for gastric cancer. Lymphovascular invasion (LVI) is a potential predictor of biological behavior. The relationship between LVI and tumor thickness (T status) has not been established in population-based studies. METHODS: Clinicopathological and survival data of 577 patients at nine centers, from between 1991 and 1997, was collected from patient records and a Provincial Cancer Registry. The primary endpoint of the study was death. A secondary analysis of a node-negative subgroup examined the significance of LVI with respect to T status. RESULTS: The population disease-specific survival was 28%. In a multivariate analysis, T, N, M, esophageal margin, tumor morphology, and residual tumor category were independent predictors of survival. LVI was documented in 58% of resected tumors. LVI correlated with advancing T and N status but was not significant in a multivariate population model. Subgroup analysis of node-negative gastric cancer found T status and LVI to be independent predictors of survival. LVI was associated with a 5-year survival of 8%, versus 43% among patients in whom it was absent (P <.001). CONCLUSIONS: T status and N status were the most important independent predictors of survival in a population-based study of gastric cancer. LVI correlated with advancing N and T status. Multivariate analysis of node-negative patients showed LVI and T status are independent predictors of survival.

Outcomes after the regionalization of major surgical procedures in the Alberta Capital Health Region (Edmonton).

OBJECTIVE: To evaluate the impact of regionalization on the outcomes of 16 surgical procedures performed in the Capital Health Region (Edmonton) of Alberta. DESIGN: A computer search of hospital discharge abstracts coded for the Canadian Institute for Health Information. SETTING: Two major hospitals in Edmonton. PATIENTS: The study population comprised 9250 patients (9727 procedures [4524, pre-regionalization, 5203 post-regionalization]) who underwent any of 16 major procedures in the 2 years before and the 2 years after restructuring. OUTCOME MEASURES: Demographic data, Charlson's comorbidity index, number of urgent and emergent cases, death rate, average length of hospital stay and the readmission rate. RESULTS: The post-regionalization patient group was slightly older, had a higher comorbidity index, and fewer urgent and emergent cases. The case volume increased by 15%, and 43.6% of patients used some form of community-based health care services. The median length of hospital stay decreased from 8.0 days pre-regionalization to 7.0 days post-regionalization (p < 0.001). Overall and for specific procedures the death rate was unchanged (3.1% pre-regionalization, 2.4% post-regionalization, p = 0.06). The readmission rates were similar for both groups (8.0% versus 7.0%). CONCLUSIONS: The consolidation of these 16 major surgical procedures had minimal impact on death and readmission rates even though patients in the post-regionalization group were slightly older and had greater comorbidity. There was a significant decline in the length of hospital stay, which occurred nationally over the same period, and a corresponding increase in the use of community-based services.

Antenatal screening by history taking--a missed opportunity.

In order to determine how effectively obstetricians and midwives identify and manage women at increased risk of fetal abnormality a case note review was undertaken. Two district general hospitals in each of two health regions and their tertiary referral centres making seven hospitals in all were studied. Women booked for antenatal care over a one-year period were included (19 895) which was estimated to be 75% of the total. Significant numbers of women at high risk because of age, past obstetric, medical or family history were not offered appropriate tests such as detailed scanning, carrier status testing, amniocentesis or chorion villus sampling. Asian women were less likely to be offered screening compared with Caucasians. Women at low risk were subjected to inappropriate tests such as amniocentesis. Ultrasound departments were rarely informed that a woman had had a previous baby with a structural abnormality. In conclusion the quality of booking histories was poor allowing important information about past medical, family or obstetric history to be overlooked. Maternity units should use carefully structured forms taking when booking histories and all staff must have regular training on risk factors.

The impact of regionalization on a surgery program in the Canadian health care system.

OBJECTIVE: To examine the impact of the regionalization of health care on the provision of surgical services in the Capital Health Region (Edmonton) of the province of Alberta. DESIGN: A 4-year retrospective descriptive analysis using data from the Canadian Institute for Health Information and from the Capital Health Region data banks. SETTING: To control health care costs, the provincially funded health care system in Alberta reformed its governance structure and service provision model. We studied community hospitals and an academic health sciences center. PATIENTS: All patients undergoing surgical care in the region. INTERVENTIONS: Regionalization of the organizational structure with the elimination of hospital boards, consolidation of services on specific sites within the regional system, and a major reduction in funding. OUTCOME MEASURES: Inpatient and day surgery procedure volumes, average length of hospital stay, relative value units, bed use, and mortality. RESULTS: The Capital Health Region has a population of 723,000 people, with 5 acute care institutions. Eighteen clinical programs now provide care through 2 referral hospitals and 3 community health centers. The reduction in operating dollars for this region was $167.1 million from fiscal years 1992-1993 to 1996-1997. Redistribution of surgical services occurred on July 1, 1995, resulting in an 18% inpatient bed reduction. Regionally, the number of acute care beds has declined from 2.25 to 1.47 per 1000 population (P < .001). Bed use has fallen from 637 to 442 inpatient days per 1000 population (P < .001). The surgery volume (1995-1996) was 44770 procedures (-3.1%). Redistribution of surgical services into high- and low-acuity settings has resulted in most surgeons working on 2 sites. Overall average length of hospital stay has decreased significantly (P < .001); however, it has increased, together with the average relative value units, in the institutions caring for patients with high-acuity surgical illnesses. Mortality remains unchanged. CONCLUSIONS: Regionalization and funding reductions within the surgical program in the Capital Health Region have resulted in a small reduction in surgical volumes. There have been major changes in service provision and the way surgeons practice.

Pulmonary resection for lung trauma.

BACKGROUND: Pulmonary resection is rarely required for trauma, and its mortality is reportedly high. METHODS: A 10-year retrospective review of pulmonary resections for trauma was done. RESULTS: Of 2,455 patients with chest trauma, 183 (7.4%) underwent thoracotomy and 32 (1.3%) required pulmonary resection. Mean age was 28.4 years and mean injury severity score was 24.5. Mechanism of injury was stab wound in 14 patients, gunshot wound in 6, and blunt trauma in 12. Blunt trauma patients had a higher injury severity score (29.6) than penetrating trauma patients (21.4), but this was not significant (p < 0.07). Indications for thoracotomy were hemorrhage in 24 patients, airway disruption in 4, and other indications in 4. Operations consisted of wedge resection (19 patients), lobectomy (9), and pneumonectomy (4). Four (12.5%) patients (pneumonectomy, 2; lobectomy, 1; wedge, 1) died. Mortality for pneumonectomy was 50%, but this was not significantly higher than for lesser resections. Blunt trauma had a higher mortality (33%) than penetrating trauma (0%) (p < 0.02). Nonsurvivors had higher injury severity scores (44.2) than survivors (21.6) (p < 0.001). CONCLUSIONS: Pulmonary resection is infrequently required for lung injury. Overall mortality is lower than previously reported, but pneumonectomy has a high mortality. Blunt trauma has a higher mortality than penetrating trauma. Injury severity scores are higher for nonsurvivors than survivors; this shows the importance of associated injuries on outcome.

Effects of isothiocyanates on cytochrome P-450 1A1 and 1A2 activity and on the mutagenicity of heterocyclic amines.

In this study we investigated the effects of phenyl isothiocyanate (PITC), benzyl isothiocyanate (BITC), phenethyl isothiocyanate (PEITC) and phenylpropyl isothiocyanate (PPITC) on the mutagenicity of five heterocyclic amines, IQ, MeiQx, Trp-P-2, Glu-P-2 and PhIP in Salmonella typhimurium strain TA 98 in the presence of liver microsomes from male Syrian golden hamsters. In addition, the effects of these isothiocyanates on cytochrome P-450 1A1 and cytochrome P-450 1A2 activities were determined by measuring the deethylation and the demethylation of ethoxyresorufin (EROD) and methoxyresorufin (MROD) respectively. With the exception of Trp-P-2, all four isothiocyanates significantly inhibited HCA-induced mutagenesis in TA 98 at concentrations of 0.05 and 0.1 mumoles/plate. BITC was the only isothiocyanate tested that showed a dose-dependent inhibition of Trp-P-2-induced mutagenesis. These four isothiocyanates showed a dose-dependent inhibition of EROD activity and, with the exception, of BITC, of MROD activity also. This indicates that the inhibition of HCA-induced mutagenesis correlates with the inhibition of cytochrome P-450 1A1 and 1A2. These P450s are known to metabolically activate HCAs. The inhibitory effects of the isothiocyanates were greater toward HCA-induced mutagenesis in TA 98 than toward EROD and MROD activity. This indicates that the antimutagenic effects of PITC, BITC, PEITC and PPITC on HCA-induced mutagenesis in Salmonella typhimurium TA 98 involves more than the inhibition of cytochrome P-450 1A1 and 1A2 activity in hamster liver microsomes.

Hemodynamic changes and gut barrier function in sequential hemorrhagic and endotoxic shock.

Multisystem organ failure (MSOF) is the major cause of late death following trauma. The gut is hypothesized to be the source of an ongoing systemic inflammatory response that drives MSOF. It has also been suggested that while a single physiologic insult might not reliably cause MSOF, the addition of a delayed second stress will. This is known as the "two-hit" theory. The purpose of this study was to investigate the two-hit theory by observing the hemodynamic and bacteriologic response to a second stress in a subacute pig model of hemorrhagic and endotoxic shock. Swine (n = 18, 30-40 kg) were fed an antibiotic-free diet for 14 days. During instrumentation and experimentation on days 1 and 3, all animals were anesthetized (ketamine, isofluorane). On day 1, all animals had placement of central venous and arterial catheters, a portal venous catheter, and superior mesenteric artery flow probe. Group E (n = 6) underwent instrumentation on day 1, then infusion of endotoxin (25 mcg/kg E. coli lipopolysaccharide) on day 3. Group HE (n = 7) underwent instrumentation then hemorrhagic shock (mean arterial pressure = 40 mm Hg for 4 hours) on day 1, then infusion of endotoxin on day 3. Group H (n = 5) were instrumented and hemorrhaged on day 1, and underwent anesthesia only on Day 3. Between periods of anesthesia the animals were allowed food and water ad lib and systemic blood was sampled for culture every 12 hours. On day 5, the animals were euthanized prior to organ sampling for bacterial culture. One animal from group HE died during endotoxic shock on day 3.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of balanol from Verticillium balanoides and ophiocordin from Cordyceps ophioglossoides.

Recently, we reported the isolation of the potent protein kinase C inhibitor balanol (1) from the fungus Verticillium balanoides. In an earlier study, König et al. reported the isolation of ophiocordin (3), a structural isomer of 1, from the fungus Cordyceps ophioglossoides. The present study was designed to clarify whether or not balanol and ophiocordin are different compounds. The results indicated that the two fungi produced the same compound, the structure being that assigned to balanol. In addition, a thirty-fold increase in the production of balanol from V. balanoides was observed when the culture medium was changed from cornmeal/tomato paste to soy meal/glycerol.

Effects of isothiocyanate alkyl chain-length on hamster liver cytochrome P-450 activity.

The tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is metabolized by various isozymes of cytochrome P-450 present in microsomes. In this study, we examined the effects of the isothiocyanate homologues, phenyl isothiocyanate (PITC), benzyl isothiocyanate (BITC), phenethyl isothiocyanate (PEITC) and phenylpropyl isothiocyanate (PPITC) on the mutagenicity and in vitro metabolism of NNK by Syrian golden hamster liver microsomes and on the in vitro microsomal metabolism of testosterone. Each isothiocyanate compound inhibited N-oxidation and alpha-hydroxylation reactions of NNK that, except for PITC, correlated with an inhibition of microsomal-mediated mutagenicity of NNK in Salmonella typhimurium TA1535. Each isothiocyanate also inhibited cytochrome P-450-mediated hydroxylation reactions of the metabolism of testosterone. In general, the inhibitory potency of the isothiocyanates corresponded with the length of the alkyl chain of the compound. Our data support the ability of isothiocyanates to inhibit the activity of a number of isozymes of cytochrome P-450.

Effects of isothiocyanates on the metabolism of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and benzo[a]pyrene by hamster and rat liver microsomes.

Benzyl isothiocyanate (BITC) and phenethyl isothiocyanate (PEITC) strongly inhibited the N-pyridine oxidation and alpha-carbon hydroxylation pathways of the in vitro metabolism of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) by Syrian golden hamster liver microsomes from saline-injected (noninduced) animals. PEITC was more potent than BITC. NNK-reduction was enhanced by both isothiocyanates. Both N-oxidation and alpha-hydroxylation activities were several-fold greater in hamster liver microsomes compared with F344 rat liver microsomes. Consequently, the inhibitory effect of BITC and PEITC on rat liver microsomes (noninduced) was not as pronounced. NNK-reduction in rat liver microsomes was not significantly different from hamster and was not enhanced by BITC and PEITC. Neither BITC nor PEITC had a strong inhibitory effect on the in vitro metabolism of benzo[a]pyrene (BaP) by either hamster or F344 rat liver microsomes from beta-naphthoflavone treated animals. The extent of BaP metabolism was similar for the two microsome groups. Since the metabolism of NNK and BaP depends upon cytochrome P450-mediated reactions that may utilize different isozymes of cytochrome P450, our data suggest that BITC and PEITC may inhibit the activity of some isozymes and not others. Our results also indicate that the inhibition of the metabolism of NNK by isothiocyanates as previously described for the mouse and rat can now be extended to include the hamster as well.

Effects of capsaicin on liver microsomal metabolism of the tobacco-specific nitrosamine NNK.

Chemically-induced mutagenesis and carcinogenesis is modulated by various plant products, some of which are present in the human diet. 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a potent carcinogen in tobacco and tobacco smoke, is activated by microsomal enzymes. In this study, we investigated the effects of capsaicin on the in vitro metabolism of NNK. Capsaicin is the principal component of Capsicum fruits used widely by humans as a food additive. Liver microsomes from saline-injected, phenobarbital-induced and beta-naphthoflavone-induced hamsters were used. Microsomes from phenobarbital and beta-naphthoflavone-induced animals expressed decreased NNK reduction and enhanced pyridine-N-oxidation, but did not significantly alter alpha-carbon hydroxylation of NNK. Capsaicin (0.5 mM) inhibited the formation of all metabolites of NNK by all microsomal fractions and inhibited alpha-hydroxylation by phenobarbital-induced microsomes more than by either of the other two treatments. Our results suggest that capsaicin, as a naturally occurring dietary constituent, possesses antimutagenic and anticarcinogenic properties through the inhibition of xenobiotic metabolizing enzymes.

Inhibition of liver microsomal cytochrome P450 activity and metabolism of the tobacco-specific nitrosamine NNK by capsaicin and ellagic acid.

The tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3- pyridyl)-1-butanone (NNK), present in tobacco and tobacco smoke, is metabolically activated by microsomal enzymes. In this study, we examined the effect of capsaicin and ellagic acid on the in vitro metabolism of NNK by hamster and rat liver microsomes. Capsaicin is the principal component of Capsicum fruits used widely throughout the world as a food additive. Ellagic acid, with reported anticarcinogenic properties, is found in various soft fruits and nuts. Both capsaicin and ellagic acid inhibited the major pathways of NNK-reduction, N-pyridine oxidation and a-hydroxylation by hamster liver microsomes. Capsaicin inhibited NNK-reduction and a-hydroxylation and ellagic acid inhibited N-oxidation and a-hydroxylation by rat liver microsomes. The effects of capsaicin and ellagic acid on isozymes of cytochrome P450 were observed in the hydroxylation reactions of the metabolism of the steroid hormone testosterone. Results of these experiments indicated that both capsaicin and ellagic acid strongly inhibited the constitutive enzymes CYP 2A2, 3A1, 2C11, 2B1, 2B2 and 2C6. This study suggests that capsaicin and ellagic acid, as naturally occurring dietary constituents, possess antimutagenic and anticarcinogenic properties through the inhibition of xenobiotic metabolizing enzymes.

A cluster of patients with inferior rectus restriction following local anesthesia for cataract surgery.

During a 5-month period, four patients developed contracture of the inferior rectus muscle following local anesthesia for cataract surgery in the practice of one ophthalmologist. Two anesthesiologists administered retrobulbar anesthesia. All patients had persistent vertical diplopia caused by a large hypotropia of the operated eye. Forced duction testing revealed marked restriction to elevation of the eye. In two patients, MRI demonstrated segmental thickening of the inferior rectus muscle, just posterior to the globe. All patients had normal thyroid function tests. No patient had a previous history of strabismus. Three of the four have had strabismus surgery consisting of recession of the markedly restricted inferior rectus muscle on an adjustable suture. These three patients have regained fusion and are now asymptomatic.

The use of blood in resuscitation of the trauma patient.

The ability to transfuse blood (a form of tissue transplantation) with relatively few immediate and long-term complications has led to increased survival in victims of injury who require massive amounts of blood. The primary deficit in hypovolemic shock secondary to trauma is in oxygen transport to the hypoperfused tissues; therefore, blood transfusion has an essential role in therapy during resuscitation and definitive treatment. The major immediate complications to be avoided are hypothermia and acidosis, which are the main causes of the coagulopathy associated with massive transfusion. The most worrisome long-term complication is the transmission of disease, of which hepatitis C is the most frequent. With improved screening techniques and heightened donor awareness, the risk of disease transmission is less than 2%. Until synthetic oxygen-carrying solutions are available, the transfusion of red blood cells, when appropriately indicated, will remain an important component in the resuscitation of the trauma patient.