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Objectives Easily administrated cyanide antidotes are needed for first responders, military troops, and emergency department staff after cyanide exposure in mass casualty incidents or due to smoke inhalation during fires involving many victims. Hydroxocobalamin has proven to be an effective antidote, but cannot be given intramuscularly because the volume of diluent needed is too large. Thus, intraosseous (IO) infusion may be an alternative, as it is simple and has been recommended for the administration of other resuscitation drugs. The primary objective of this study was to compare the efficacy of IO delivery of hydroxocobalamin to intravenous (IV) injection for the management of acute cyanide toxicity in a well-described porcine model. Methods Twenty-four swine (45 to 55 kg) were anesthetized, intubated, and instrumented with continuous mean arterial pressure (MAP) and cardiac output monitoring. Cyanide was continuously infused until severe hypotension (50% of baseline MAP), followed by IO or IV hydroxocobalamin treatment. Animals were randomly assigned to receive IV (150 mg/kg) or IO (150 mg/kg) hydroxocobalamin and monitored for 60 minutes after start of antidotal infusion. The primary outcome measure was the change in MAP after antidotal treatment from onset of hypotension (time zero) to 60 minutes. A sample size of 12 animals per group was determined by group size analysis based on power of 80% to detect a one standard deviation of the mean MAP between the groups with an alpha of 0.05. Whole blood cyanide, lactate, pH, nitrotyrosine (nitric oxide marker) levels, cerebral and renal near infrared spectrometry (NIRS) oxygenation, and inflammatory markers were also measured. Repeated-measures analysis of variance was used to determine statistically significant changes between groups over time. Results At baseline and at the point of hypotension, physiologic parameters were similar between groups. At the conclusion of the study, 10 out of 12 animals in the IV group and 10 out of 12 in IO group survived (p = 1.0). Both groups demonstrated a similar return to baseline MAP (p = 0.997). Cardiac output, oxygen saturation, and systemic vascular resistance were also found to be similar between groups (p > 0.4), and no difference was detected between bicarbonate, pH, and lactate levels (p > 0.8). Cyanide levels were undetectable after the hydroxocobalamin infusion throughout the study in both groups (p = 1.0). Cerebral and renal NIRS oxygenation decreased in parallel to MAP during cyanide infusion and increased after antidote infusion in both groups. Serum nitrotyrosine increased during cyanide infusion in all animals and then decreased in both study arms after hydroxocobalamin infusion (p > 0.5). Serum cytokines increased starting at cyanide infusion and no difference was detected between groups (tumor necrosis factor-α, interleukin [IL]-1β, IL-6, and IL-10). Conclusions The authors found no difference in the efficacy of IV versus IO hydroxocobalamin in the treatment of severe cyanide toxicity in a validated porcine model. Resumen Objetivos se necesitan antidotos para el cianuro de facil administracion para los primeros respondedores, las tropas militares y el personal del servicio de urgencias tras una exposicion al mismo en grandes cantidades o debido a la inhalacion de humo durante fuegos que involucren a muchas victimas. La hidroxocobalamina ha demostrado ser un antidoto efectivo, pero no puede ser administrada por via intramuscular debido a que el volumen de disolvente necesario es demasiado grande. Por ello, la via intraosea (IO) puede ser una alternativa, por su sencillez, y ha sido recomendada para la administracion de otros farmacos durante la resucitacion. El objetivo principal del estudio fue comparar la eficacia de la administracion IO de hidroxocobalamina con la inyeccion IV para el manejo de la intoxicacion aguda por cianuro en un modelo porcino bien descrito. Metodologia Veinticuatro cerdos (45 a 55 kg) se anestesiaron, intubaron e instrumentalizaron con monitorizacion continua de la presion arterial media (PAM) y el gasto cardiaco. El cianuro se perfundio de forma continua hasta la hipotension grave (50% de la PAM basal) seguido por el tratamiento con hidroxocobalamina IO o IV. Los animales fueron asignados de forma aleatorizada a recibir hidroxocobalamina por via IV (150 mg/kg) o IO (150 mg/kg) y fueron monitorizados durante 60 minutos tras el inicio de la infusion del antidoto. La medida de resultado principal fue el cambio en la PAM tras el tratamiento con el antidoto desde el inicio de la hipotension (tiempo cero) hasta los 60 minutos. Se determino un tamano muestral de 12 animales por grupo basandose en un poder del 80% para detectar una desviacion estandar de la media de PAM entre los grupos con un error alfa de 0,05. Se midio tambien el cianuro en sangre, lactato, pH, niveles nitrotirosina (marcador de oxido nitrico), la oxigenacion cerebral y renal mediante un espectrometro de infrarrojo cercano (NIRS, near infrared spectrometry) y marcadores de inflamacion. Se utilizo el analisis de la varianza para medidas repetidas para determinar los cambios estadisticamente significativos entre los grupos con el paso del tiempo. Resultados Basalmente y en el momento de la hipotension, los parametros fisiologicos fueron similares entre los grupos. Al final del estudio, 10 de los 12 animales en el grupo IV y 10 de los 12 en el grupo IO sobrevivieron (p = 1.0). Ambos grupos demostraron un retorno similar a la PAM basal (p = 0,997). El gasto cardiaco, la saturacion de oxigeno y la resistencia vascular sistemica se encontraron tambien similares entre los grupos (p > 0,4); y no se detectaron diferencias entre los niveles de bicarbonato, pH y lactato (p > 0,8). Las concentraciones de cianuro fueron indetectables tras la infusion de hidroxocobalamina durante el estudio en ambos grupos (p = 1.0). La oxigenacion cerebral y renal mediante espectroscopia funcional cercana al infrarrojo disminuyo de forma paralela a la PAM durante la infusion de cianuro y se incremento tras la infusion del antidoto en ambos grupos. La nitrotirosina serica se incremento durante la infusion de cianuro en todos los animales y despues descendio en ambos brazos del estudio tras la infusion de hidroxicobalamina (p > 0,5). Las citoquinas sericas se incrementaron al iniciar la infusion de cianuro y no se detecto diferencia entre los grupos (TNF-α, IL-1β, IL-6, y IL-10). Conclusiones No se encontraron diferencias en la eficacia de la hidroxocobalimina intravenosa frente a la intraosea en el tratamiento de la intoxicacion grave por cianuro en un modelo porcino validado.

Introduzione: I papillomavirus umani (HPV) sono piccoli virus a DNA, suddivisi vari genotipi, con rischio di trasformazione oncogena per integrazione del DNA virale nel DNA cellulare; nello studio abbiamo cercato di discriminare i singoli genotipi di HPV in una popolazione salentina a rischio, afferente al Laboratorio di Biologia Molecolare e Oncologia Sperimentale (LBMOS) del Presidio Ospedaliero “Vito Fazzi” di Lecce. Materiali e metodi: Durante il biennio 2006-2007, sono stati analizzati 816 campioni di muco cervico-vaginale di donne con sospetto di HPV, o recente/pregresso episodio di infezione, con amplificazione di ORF L1. Con sequenziamento e ibridazione con sonde, siamo andati a ricercare ogni specifico genotipo di HPV presente nella popolazione a rischio. Risultati: Analizzando 816 campioni, l’HPV è risultati essere presente in 183 casi (22.4%), con un trend di crescita dal 19%del 2006 al 27%del 2007; la ricerca dei genotipi ha prodotto 37 genotipi differenti di HPV e, fra questi, i casi con maggiore frequenza sono stati quelli di HPV-16 con 37 casi (13,81%), seguiti dall’HPV-6 con 27 casi (10.07%), dall’HPV-53 con 25 casi (9.33%) e, infine, HPV-31 con 21 casi (7.84%). Successivamente apparivano, con percentuali inferiori, il genotipo 53 (MR-HPV), il genotipo 31 e il genotipo 18 (HR-HPV), poi il genotipo 66 (MRHPV), il genotipo 58 (HR-HPV) e il ceppo 61 (MR-HPV). Conclusioni: Il sequenziamento molecolare risulta il “Gold Standard” nella ricerca dell’HPV; tale metodica ha individuato, nel territorio salentino, una maggiore presenza di HPV-16 seguito, in misura ridotta, dall’HPV-6 e da altri ceppi. Tali percentuali dimostrano una specifica realtà epidemiologica e, probabilmente, tali dati, se implementati, potrebbero rivelare una realtà differente rispetto a quella della letteratura scientifica, utile per programmare la profilassi vaccinica e la realizzazione di nuovi vaccini mirati.

Partially delactosated and demineralized whey protein powders were purified from pooled milks of individual cows homozygous for the β -lactoglobulin ( β -Lg) genetic variants A and B ( whey A and whey B). The protein composition of the two samples was similar. The two whey protein powders were dissolved in water at different pH (2, 4.5, 6.8) and NaCl concentrations (150 and 400 mmol/dm 3 ), and emulsions were made utilizing grape-seed oil. Emulsion stability (ES), emulsion activity index (EAI) and emulsion capacity (EC) were examined. The results showed the influence of the two genetic variants of β -Lg on the functional behaviour of whey. Emulsion obtained from B powder solutions showed the higher stability at the different test conditions. The best results occurred at pH 6.8 with no significant differences between the two tested NaCl concentrations. At pH2 and 400 mmol/dm 3 NaCl, both samples did not demonstrate any stability. Data on emulsion capacity indicated that whey B is less affected by the different physicochemical conditions tested (pH, NaCl concentration, heat treatment at 90°Cl5 min), while whey A gave the highest and the lowest EC values depending on the experimental conditions.

Reaction of the sterically demanding ligand precursor [(CHBut)2(C5H2But2)2]K2 with SnCl2 in THF gave the air-stable ansa-stannocene complex rac[(CHBut)2(C5H2But2)2]Sn. A single-crystal X-ray structure of the ansa-stannocene revealed a C2-symmetric and slipped bent structure, with the Sn2+ displaced toward an η2 bonding mode between bridged, highly tilted, and staggered 2,4-di-tert-butylcyclopentadienyl rings.

We sought to compare the flow rates of the proximal tibia, the distal femur, and the proximal humerus using high pressure (greater than 300 mmHg) through an intraosseous (IO) infusion needle in an adult swine model.We performed a prospective interventional study in 11 swine. After placement of central vein and arterial lines, blood was removed via the central line until the animal's mean arterial pressure decreased 25% from the recorded baseline. We inserted a 25-mm IO needle into the proximal tibia and 45-mm needles into the distal femur and proximal humerus. All extremities were utilized in each study animal. We infused normal saline at each site for 10 minutes with a pressure bag inflated to the highest achievable pressure (greater than 300 mmHg) as measured at the infusion site with a calibrated portable inline pressure gauge. We measured the volume of normal saline remaining and we calculated infusion rates for each site. We then compared infusion flow rates between the three locations. Statistical analysis and comparison of the infusion rates of all three study arms were as performed using analysis of variance (ANOVA).The average weight of the swine was 71 kg (range 64-84 kg). Successful placement of the IO needle was confirmed at all three sites. The mean infusion flow rate was 213 mL/min (standard deviation [SD] 53.2 mL/min) for the proximal humerus, 138 mL/min (SD 65.3 mL/min) for the distal femur, and 103 mL/min (SD 48.1 mL/min) for the proximal tibia (p0.001). The flow rate through the proximal humerus was statistically greater than that for the proximal tibia and the distal femur (p0.001). The flow rates through the proximal tibia and distal femur were similar.The humerus is a suitable alternative site for IO placement, with a potential for higher flow rates than the proximal tibia and distal femur when resuscitating a patient.

It was previously demonstrated that some plant inhibitors are able to inactivate the bacterial proteinase subtilisin ; since information concerning the effect of plant inhibitors on other microbial proteinases remains limited, we decided to determine the activity of the soybean Kunitz and Bowman-Birk inhibitors (KI and BBI, respectively) on 14 proteinases of fungal and bacterial origin. The results show that microbial proteinases are frequently inhibited by KI and BBI and that proteinases with the same EC number (eg subtilisin Carlsberg and subtilisin BPN') may equally give different responses to the inhibitors. In particular, all serine proteinases examined were affected by both KI and BBI while metalloproteinases were not. Inhibition was also achieved on a range of microbial proteinases for which the mechanistic class is yet to be established ; the data suggest that they belong to the serine type. In one instance, activation instead of inhibition was noted.

Objectives: Our objective was to assess patients understanding of emergency department (ED) wait times and why patients may leave the waiting room before seeing a provider. Methods: Survey of patients in the ED waiting room of an urban tertiary care military hospital where civilian and military patients are treated. Results: A total of 508/517 surveys (98%) were completed. Age ranges were 18 to 35 years (49%), 36 to 60 (31%), or older than 60 (20%). Education levels were high school (20%), some college (37%), or college graduate (39%). Of 503 respondents, 125 (25%) had left an ED waiting room before seeing a provider. The reasons included excessive wait times (91%) and family responsibilities (5%). Five hundred eight reported the factors that would motivate them to wait to see the physician (not leave without being seen [LWOBS]) were the severity of illness (64%), and if they received an update of wait times (26%); 82% (391/480) understood that severely ill patients were seen first. Patients attributed long wait times to doctors and nurses caring for other patients (292/583, 50%) and insufficient physician and nurse staffing (245, 42%). Of 802 responses for ideas to improve the wait, 34% said regular updates on estimated wait times, 21% said television shows or movies to view, 20% said books and magazines to read, and 11% said computers to access. Conclusions: Long wait times were the primary reason that patients left before seeing a provider, despite having ready access to care. Respondents attributed long wait times to patient volume and inadequate staffing. Regular updates on wait times and material for entertainment may improve the waiting experience and reduce LWOBS.

Objectives: In August 2007, the Food and Drug Administration (FDA) released a public health advisory recommending that over-the-counter cough and cold medications (CCMs) not be used in children younger than 2 years. Our objective was to assess parents awareness and understanding of the guidelines. Methods: We surveyed caregivers of children younger than 2 years in the emergency department of an urban tertiary care military hospital where civilian patients are also treated. After completing the survey, caregivers received a handout explaining the FDA s recommendations. Results: Our response rate was 99% (264/265). First-time parents constituted 45% (114/251) of responders. Education level was high school, 21%; some college, 36%; and college graduate, 40%. Thirty-one percent (77/247) were aware of the FDA guidelines. Of these 77, 44 (57%) reported the guidelines indicated CCMs were not safe in children younger than 2 years, and 18 (23%) saidCCMs have caused death. Twenty-six percent (68/264) did not give CCMs to their children younger than 2 years, and 40% of these reported it was because of learning about the guidelines; 63% (165/264) reported CCMs were effective, 11% ineffective, and 27% did not know. Fifty-seven percent (151/263) reported CCMs were safe, 12% unsafe, and 31% unsure. Twenty-two percent (31/143) planned to use or continue to use CCMs in their children, 34% did not, 23% not sure, and 21% only if their doctor advised it. Conclusions: The majority of caregivers were not aware of the FDA guidelines on CCM use in children younger than 2 years. Most thought CCMs were safe and effective.

To compare the rerum to baseline of mean arterial blood pressure (MAP) between 2 groups of swine in acute cyanide toxicity and treated with IV HOC or 10 HOC. We also compared blood cyanide, lactate, pH, nitrotyrosine levels, cerebral oxygenation, and inflammatory markers. 24 swine (48-52kg) were intubated, anesthetized, and instrumented (MAP and cardiac output (CO) monitoring). Cyanide was continuously infused until severe hypotension (50 per cent of baseline MAP). Animals were randomly assigned to IV HOC 150mg/kg or 10 HOC 15 0mg/kg and monitored for 60 min after start of antidote. Baseline mean weight, time to hypotension, and cyanide dose at hypotension were similar. At hypotension, mean MAP, blood cyanide, and lactate levels were similar. Both groups had similar return to baseline MAP. Intraosseous hydroxocobalamin led to similar return to baseline of MAP as intravenous. Mortality, heart rate, cardiac output, lactate, nitrotyrosinc, cerebral NTRS , and inflammatory markers were also similar. TO HOC may be as effective as IV HOC in acute, severe cyanide toxicity.

Solutions of D-fructose and sucrose were heated, alone and in the presence of glycine. The reaction products were examined in terms of their effects on peroxidase (POD) activity. Compared with glucose mixtures, fructose products had a great inhibiting effect on POD activity, while in the case of Maillard reaction products (MRPs), the differences in antibrowning activity between fructose-glycine and glucose-glycine mixtures were slight. As regards the sucrose and sucrose-glycine solutions, a slight inhibiting effect on POD activity was observed only after sugar hydrolysis. Measurements of the reducing power of sugar and sugar-amino acid solutions showed a correspondence between antioxidant properties of caramelization and Maillard reaction products and their antibrowning effects.

A complex chemical transformation known as the Maillard reaction may occur in foods containing reducing sugars and compounds with free amino groups. This reaction is accelerated by heating but occurs also at room temperature. Some food flavors, colors and texture characteristics depend on this reaction, but reduced nutritive values of proteins are also reported. To evaluate the contribution of digestive enzymes inhibition to this effect, activities of some proteases (pepsin, trypsin and chymotrypsin) were tested in the presence of glucose-glycine solutions heated at 90C for different times. Pepsin was partially inhibited by glycine itself but Maillard reaction products (MRP) did not produce additional effect. MRP showed a limited inhibition on trypsin and chymotrypsin. Unfractionated MRP solution showed an inhibiting effect on trypsin of the same order as already reported by Oeste (1989) for the low molecular weight (LMW) fraction of glucose-lysine MRP. High molecular weight (HMW) MRP presented an additional inhibiting effect, beside that reported for LMW on chymotrypsin. This however only occurred in particular conditions.

Background Cyanide is a common toxin in structural fires and a salt that is ingested for suicide. However, most studies have focused on the effects of inhaled cyanide. The objective of this study was to describe the incidence of cyanide ingestions, symptoms, cardiac arrest and antidotal therapy used as reported to all US poison centres over 7 years. Methods A retrospective review of cases over 7 years as reported to 61 poison centres in the USA was performed. Sole ingestions of cyanide were identified. A trained reviewer used a standard data collection sheet within a secured electronic database. Age, intent, clinical effects, treatments, antidotes and outcomes were recorded. One investigator audited a random sample of charts. Results Out of 1741 exposures, 435 ingestions were identified. Most were male (68%) and the mean age was 34 years (range 1 month–83 years). 45% of cases were intentional, most commonly as a suicide attempt. 8.3% of cases died and 9% (38/435) of patients had cardiac arrest or hypotension. 13% of all cases and 26% of cases arriving at a healthcare facility received an antidote. In 35% of cases of cardiac arrest or hypotension, and in 74% of intentional ingestions, antidotes were not given. Conclusions Suicide attempt was the most common reason for cyanide ingestion. Most of these patients died. Cardiac arrest or hypotension was common, but antidote use was not, particularly in critically ill patients. Research is needed to improve outcomes of cyanide-induced hypotension and cardiac arrest and to reduce barriers to antidote use.

Polyethylene glycols (PEGs) of various chain length were used to crosslink lysozyme onto an insoluble support such as oxirane. A very high degree of modification and no inactivation of lysozyme were obtained with PEG 20000, but enzymatic activity increased up to 20 times at pH 3.0, at which point the activity of the native enzyme was lower when using Leuconostok oenus as a macromolecular substrate.

A bacterial species (Leuconostc oenos), well suited to acidic pH values, is proposed as substrate for the spectrophotometric determination of lysozyme activity. The principal advantage of this organism as the substrate is the ease of investigating lysozyme behavior at acidic pH. The sensitivity is lower than with Micrococcus luteus, but data are highly reproducible.

The effect of glucose, fructose and sucrose aqueous solutions on polyphenol oxidase (PPO) and peroxidase (POD) activity was studied. Solutions with high concentrations of sugars showed a slight inhibiting effect on PPO while an activation was observed for POD. Glucose solutions, tested after sufficient heat treatments, showed an inhibiting action only on PPO activity. Maillard reaction products, obtained by heating a glucose/glycine solution, caused an unexpectedly strong inhibiting effect on both enzymes. The inhibition became more evident with increasing heating times of the glucose/glycine solution, reducing the activity of PPO and POD nearly to zero. Thus, the Maillard reaction products, already known for their antioxidant properties, were also shown to have a strong inhibiting effect on enzymatic browning.

Many assay procedures have been devised to measure lipolytic activity, but none is without problems. It is for this reason that new methods are still being proposed. In this work we have investigated the use of two esters of p-nitrophenol, the palmitic acid and lauric acid esters, as substrates for a highly sensitive high-performance liquid chromatographic method. Data on recovery, specific activity and reproducibility are reported only for the lauric ester, because the palmitic ester turned out to be a very poor substrate.

Two serine-proteases (P I and P II ) were isolated from the larval midgut of Spodoptera littoralis by affinity chromatography. These hydrolysed casein and BApNA and were strongly inhibited by TLCK. On SDS-PAGE, an apparent molecular weight of 27 kDa was determined for both proteases. When mass spectrometric analysis was performed directly on the eluted fractions, one main component per peak was observed, having a molecular mass of 24,359±2 Da (P I ) and 24,448±2 Da (P II ). The enzymes had their maximum activity and stability at pH 10 and 4, respectively. P I was significantly more active than P II at 20°C or less; it was also more resistant to denaturation by heat (55 versus 50°C). A breakpoint between activity and stability was recorded at 30°C for both proteases. P II was strongly inhibited by aprotinin, leupeptin and KI, and partly inactivated by LBTI and chymostatin; P I was unaffected by LBTI and chymostatin and was also less susceptible to aprotinin and KI. On the basis of substrate degradation and inactivation rates, it was concluded that P I and P II are very different from the two proteases found in another strain of S. littoralis . Though P I and P II were found to belong to the same mechanistic class of bovine trypsin, they were not activated by Ca 2+ over the concentration range 0–20 mM. It is suggested that transgenic plants harbouring genes coding for effective P I inhibitors should have a high level of resistance towards this insect pest.

Autoimmune endocrinopathies are characterised by an increased number of peripheral blood lymphocytes (PBL) expressing activation/ memory markers on their surface. The aim of this study was to determine whether a similar finding could be detected in a group of 11 paediatric and young adult patients suffering from autoimmune polyglandular syndrome type 1 (APS1), also called autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), as very few data have previously been reported in this field. The control group was made up of 11 sex- and age-matched healthy subjects. Fifteen lymphocyte subsets were compared, in terms of percentage and absolute number, and statistical analysis was performed by the Mann-Whitney test. Measurement of T (CD3+), B (CD19+), natural killer (NK, CD3-CD16/56+), CD4+ and CD8+ T lymphocytes showed that patients with APS1 had a higher percentage and absolute count of T lymphocytes: this was entirely due to the statistically larger CD3+CD4+ fraction. Patients with APS1 also had slightly fewer B and NK lymphocytes, but the difference was negligible. Comparison of CD4+ subpopulations bearing activation and naive/memory antigens (marked by CD69, CD25, anti-HLA-DR, CD45RA and CD45RO) showed that patients with APS1 had generally larger percentages and absolute counts of these subsets: however, only the percentage and absolute size of the CD4+CD25+ subset (p = 0.0354 and p = 0.0151, respectively), and the absolute number of the CD4+ anti-HLA-DR+ and CD4+ CD45RO+ subsets (p = 0.0193 and p = 0.0209, respectively) were significantly higher. Interestingly, patients with APS1 also had significantly fewer CD8+CD11b+ and CD3-CD8+ cells. In conclusion, PBL distribution in APS1 resembles that of other autoimmune diseases. Further studies are needed to confirm and possibly extend these data.

Many biochemical and molecular details are available for soybean seed proteinase inhibitors, but little is known about the quantitative regulation of structural genes. In order to fill this gap, a complete set of diallel crosses was made between inbred lines characterised by the same inhibitor array coupled with a different inhibitor content in the raw flour. The Hayman analysis revealed the presence of different regulatory elements in the parental lines, giving rise to both additive and dominance variation. Dominant alleles were found to decrease the trypsin inhibitory activity (TI activity) shown by the raw flours and appeared to have higher frequencies than recessive alleles. Significant maternal effects were also detected, particularly in crosses of low-TI activity female x high-TI activity male; maternal effects strengthened the role of the regulatory genes transferred by the female parent to the hybrid. Data fitted the simple additive-dominance model with genes independent in both action and distribution. Narrow and broad heritability values were 54% and 82% respectively, thus indicating the feasibility of lowering TI activity through selection.