Inactivation of Escherichia coli O1 57:H7, Listeria monocytogenes, and Lactobacillus leichmannii by combinations of ozone and pulsed electric field.

Pulsed electric field (PEF) and ozone technologies are nonthermal processing methods with potential applications in the food industry. This research was performed to explore the potential synergy between ozone and PEF treatments against selected foodborne bacteria. Cells of Lactobacillus leichmannii ATCC 4797, Escherichia coli O157:H7 ATCC 35150, and Listeria monocytogenes Scott A were suspended in 0.1% NaCl and treated with ozone, PEF, and ozone plus PEE Cells were treated with 0.25 to 1.00 microg of ozone per ml of cell suspension, PEF at 10 to 30 kV/cm, and selected combinations of ozone and PEF. Synergy between ozone and PEF varied with the treatment level and the bacterium treated. L. leichmannii treated with PEF (20 kV/cm) after exposure to 0.75 and 1.00 microg/ml of ozone was inactivated by 7.1 and 7.2 log10 CFU/ml, respectively; however, ozone at 0.75 and 1.00 microg/ml and PEF at 20 kV/cm inactivated 2.2, 3.6, and 1.3 log10 CFU/ml, respectively. Similarly, ozone at 0.5 and 0.75 microg/ml inactivated 0.5 and 1.8 log10 CFU/ml of E. coli, PEF at 15 kV/cm inactivated 1.8 log10 CFU/ml, and ozone at 0.5 and 0.75 microg/ml followed by PEF (15 kV/cm) inactivated 2.9 and 3.6 log10 CFU/ml, respectively. Populations of L. monocytogenes decreased 0.1, 0.5, 3.0, 3.9, and 0.8 log10 CFU/ml when treated with 0.25, 0.5, 0.75, and 1.0 microg/ml of ozone and PEF (15 kV/cm), respectively; however, when the bacterium was treated with 15 kV/cm, after exposure to 0.25, 0.5, and 0.75 microg/ml of ozone, 1.7, 2.0, and 3.9 log10 CFU/ml were killed, respectively. In conclusion, exposure of L. leichmannii, E. coli, and L. monocytogenes to ozone followed by the PEF treatment showed a synergistic bactericidal effect. This synergy was most apparent with mild doses of ozone against L. leichmannii.

Novel quantitative assays for estimating the antimicrobial activity of fresh garlic juice.

Novel agar diffusion and broth dilution assays were developed for quantitatively estimating the antimicrobial activity of fresh garlic juice. Bacteria found to be inhibited by garlic juice in agar diffusion assay included two gram-positive and five gram-negative species. Leuconostoc mesenteroides was not inhibited. Escherichia coli B-103 (HB101, with pJH101, ampicillin resistant, 100 microg ml(-1)) was inhibited and chosen as the standard culture for quantitative assays. The agar diffusion assay was based on the slope ratio method, where the slope of dose response for garlic juice was divided by the slope of dose response for methylmethane thiosulfonate (MMTSO2). Juice from fresh garlic varied in activity between 1.76 and 2.31 microg of MMTSO2 per mg of garlic juice. The activity of juice decreased during 11 months of storage of garlic cloves at 5 degrees C from 2.31 to less than 0.1 microg of MMTSO2 per mg of juice. The broth dilution assay also used the E. coli B-103 culture, which permitted selective enumeration of this bacterium when 100 microg ml(-1) of ampicillin was incorporated into the enumerating agar. Selective enumeration was essential since the garlic juice was not sterile and, thus, contained natural flora. Growth of E. coli was unaffected by 0.1%, delayed by 0.25%, and completely inhibited at 0.5 and 2% garlic juice in broth during 24 h of incubation at 37 micro C. The minimum inhibition concentration of garlic juice by broth dilution assay was, thus, estimated to be 0.5%, which is equivalent to 3.46 microg of MMTSO2 per mg of garlic juice by the agar diffusion assay.

Effects of otitis media with effusion on the vestibular system in children.

Middle ear effusion has been considered the most common cause of vestibular disturbance in children. However, there have been only a few studies on vestibular disturbance in children with otitis media with effusion. We studied the vestibular systems of 30 children with otitis media with effusion aged 8 to 13 years and compared the results with 15 age- and sex-matched controls. A questionnaire relating to vestibular disturbance was given to patients and their parents. Spontaneous nystagmus and positional nystagmus were recorded by electronystagmography as diagnostic tests of the vestibular system. Romberg's and past-pointing tests were performed on children with otitis media with effusion and controls. After vestibular tests were completed, myringotomy was performed, and a ventilation tube was inserted. The questionnaire and the vestibular tests were repeated after the operation and during the first month after surgery. Our study showed that there was a history of vestibular disturbance in 33% of children with otitis media with effusion. Electronystagmography and Romberg's test findings demonstrated that 33% of the children had vestibular dysfunction (p < 0.05). After myringotomy with ventilation tube insertion, vestibular test results returned to normal, and symptoms related to vestibular disturbance improved. These findings confirm the assumption that middle ear effusion may affect the vestibular system, which can be resolved after myringotomy with ventilation tube insertion.

The advantages of MRI and MRA for diagnosing Behçet's disease and internal jugular vein thrombosis.

Behçet's disease is a multisystem disorder characterized by recurrent aphtous ulcers of mucosal membranes, skin lesions, and vasculitis. The prevalence of vascular involvement is about 25% and this is the leading cause of death in Behçet's disease. Jugular vein involvement is an unusual manifestation of Behçet's disease. Medical insertion of needles into veins or arteries can induce thrombosis or aneurysms. Thus, diagnostic studies or therapy can aggravate the disease and even be fatal. We report a patient with Behçet's disease and rare internal jugular vein thrombosis. We also demonstrate the ability of noninvasive magnetic resonance imaging and magnetic resonance angiography to aid in the diagnosis and evaluation of vascular involvement in Behçet's disease, without risking iatrogenic complications.

[40-year-old patient with an aneurysm of the extracranial arteria carotis interna]. / 40-jährige Patientin mit einem Aneurysma der extrakraniellen Arteria carotis interna.

A case of extracranial internal carotid artery aneurysm Aneurysms of the extracranial internal carotid artery are uncommon but can be responsible for serious complications such as rupture, thrombosis, or embolism. The underlying causes of aneurysm include atherosclerosis, fibromuscular dysplasia, trauma, previous carotid artery surgery, infection, dissection, syphilis and a congenital defect. Five distinct clinical types of aneurysms are reported with different symptoms and treatment. We report a case of extracranial internal carotid artery aneurysm and its successfully surgical treatment.

Giant polypoid tumor of the esophagus.

A patient with a giant polypoid tumor of the esophagus, measuring 22 cm in length is described in this report. The patient presented with cough attacks and respiratory distress. Diagnostic and therapeutic intervention required aggressive airway management, radiographic and endoscopic evaluation, and definitive surgical treatment. Benign esophageal tumors are rarely seen and originate from the upper third of esophagus, frequently close to the cricopharyngeus muscle. They may attain giant proportions. A variety of clinical presentations are described, the most serious being asphyxia secondary to laryngeal obstruction. We observed a giant esophageal tumor which was interpreted as angiofibromyolipoma that caused laryngeal obstruction. We present the clinical picture and histopathological findings of the tumor.

Modeling of WalkMECH: a fully-passive energy-efficient transfemoral prosthesis prototype.

In this paper we present the port-based model of WalkMECH, a fully-passive transfemoral prosthesis prototype that has been designed and realized for normal walking. The model has been implemented in a simulation environment so to analyze the performance of the prosthetic leg in walking experiments and so to enhance the mechanics of the system. The accuracy of the model has been validated by experimental tests with a unilateral amputee participant.

Design of a fully-passive transfemoral prosthesis prototype.

In this study, we present the mechanical design of a prototype of a fully-passive transfemoral prosthesis for normal walking. The conceptual working principle at the basis of the design is inspired by the power flow in human gait, with the main purpose of realizing an energy efficient device. The mechanism is based on three elements, which are responsible of the energetic coupling between the knee and ankle joints. The design parameters of the prototype are determined according to the human body and the natural gait characteristics, in order to mimic the dynamic behavior of a healthy leg. Hereby, we present the construction details of the prototype, which realizes the working principle of the conceptual mechanism.

CD36 as a multiple-ligand signaling receptor in atherothrombosis.

The glycoprotein CD36, also known as glycoprotein IIIb/IV or FAT, is expressed on the surface of platelets, monocytes, microvascular endothelial cell, smooth muscle cells, cardiomyocytes and other cells of the cardiovascular system. In spite of its abundant presence, CD36 has remained for long a mysterious protein with a poorly understood role. In this paper, we review how CD36 can affect cellular responses by interaction with a variety of ligands, in particular thrombospondin-1, oxidized lipoproteins and fatty acids. Furthermore, given the structure of CD36 with two transmembrane domains and short cytoplasmic tails, we consider how this receptor can induce intracellular signaling, likely in junction with other cellular receptors or associated proteins in the membrane. Current literature points to activation of Src-family and mitogen-activated protein kinases, as well as to activation of the NFκB and Rho pathways. The new insights make CD36 attractive as a therapeutic target to suppress platelet and monocyte/macrophage function and thereby atherothrombosis.

Signaling role of CD36 in platelet activation and thrombus formation on immobilized thrombospondin or oxidized low-density lipoprotein.

BACKGROUND AND OBJECTIVE: Platelets abundantly express glycoprotein CD36 with thrombospondin-1 (TSP1) and oxidized low-density lipoprotein (oxLDL) as proposed ligands. How these agents promote platelet activation is still poorly understood. METHODS AND RESULTS: Both TSP1 and oxLDL caused limited activation of platelets in suspension. However, immobilized TSP1 and oxLDL, but not LDL, strongly supported platelet adhesion and spreading with a major role of CD36. Platelet spreading was accompanied by potent Ca(2+) rises, and resulted in exposure of P-selectin and integrin activation, all in a CD36-dependent manner with additional contributions of α(IIb) ß(3) and ADP receptor stimulation. Signaling responses via CD36 involved activation of the protein tyrosine kinase Syk. In whole blood perfusion, co-coating of TSP1 or oxLDL with collagen enhanced thrombus formation at high-shear flow conditions, with increased expression on platelets of activated α(IIb) ß(3), P-selectin and phosphatidylserine, again in a CD36-dependent way. CONCLUSIONS: Immobilized TSP1 and oxLDL activate platelets partly via CD36 through a Syk kinase-dependent Ca(2+) signaling mechanism, which enhances collagen-dependent thrombus formation under flow. These findings provide novel insight into the role of CD36 in hemostasis.

Biomechanical conceptual design of a passive transfemoral prosthesis.

In this study, we present the conceptual design of a fully-passive transfemoral prosthesis. The proposed design is inspired by the analysis of the musculo-skeletal activity of the healthy human leg. In order to realize an energy efficient device, we introduce three storage elements, which are responsible of the energetic coupling between the knee and the ankle joints. Simulation results show that the power storage of the designed conceptual prosthesis is comparable with the human gait.

Complementary roles of platelets and coagulation in thrombus formation on plaques acutely ruptured by targeted ultrasound treatment: a novel intravital model.

BACKGROUND: Atherothrombosis is a major cause of cardiovascular events. However, animal models to study this process are scarce. OBJECTIVES: We describe the first murine model of acute thrombus formation upon plaque rupture to study atherothrombosis by intravital fluorescence microscopy. METHODS: Localized rupture of an atherosclerotic plaque in a carotid artery from Apoe(-/-) mice was induced in vivo using ultrasound. Rupture of the plaque and formation of localized thrombi were verified by two-photon laser scanning microscopy (TPLSM) in isolated arteries, and by immunohistochemistry. The thrombotic reaction was quantified by intravital fluorescence microscopy. RESULTS: Inspection of the ultrasound-treated plaques by histochemistry and TPLSM demonstrated local damage, collagen exposure, luminal thrombus formation as well as intra-plaque intrusion of erythrocytes and fibrin. Ultrasound treatment of healthy carotid arteries resulted in endothelial damage and limited platelet adhesion. Real-time intravital fluorescence microscopy demonstrated rapid platelet deposition on plaques and formation of a single thrombus that remained subocclusive. The thrombotic process was antagonized by thrombin inhibition, or by blocking of collagen or adenosine diphosphate receptor pathways. Multiple thrombi were formed in 70% of mice lacking CD40L. CONCLUSIONS: Targeted rupture of murine plaques results in collagen exposure and non-occlusive thrombus formation. The thrombotic process relies on platelet activation as well as on thrombin generation and coagulation, and is sensitive to established and novel antithrombotic medication. This model provides new possibilities to study atherothrombosis in vivo.

Plasma serotonin levels and the platelet serotonin transporter.

Serotonin (5HT) is a platelet-stored vasoconstrictor. Altered concentrations of circulating 5HT are implicated in several pathologic conditions, including hypertension. The actions of 5HT are mediated by different types of receptors and terminated by a single 5HT transporter (SERT). Therefore, SERT is a major mechanism that regulates plasma 5HT levels to prevent vasoconstriction and thereby secure a stable blood flow. In this study, the response of platelet SERT to the plasma 5HT levels was examined within two models: (i) in subjects with chronic hypertension or normotension; (ii) on platelets isolated from normotensive subjects and pretreated with 5HT at various concentrations. The platelet 5HT uptake rates were lower during hypertension due to a decrease in Vmax with a similar Km; also, the decrease in Vmax was primarily due to a decrease in the density of SERT on the platelet membrane, with no change in whole cell expression. Additionally, while the platelet 5HT content decreased 33%, the plasma 5HT content increased 33%. Furthermore, exogenous 5HT altered the 5HT uptake rates by changing the density of SERT molecules on the plasma membrane in a biphasic manner. Therefore, we hypothesize that in a hypertensive state, the elevated plasma 5HT levels induces a loss in 5HT uptake function in platelets via a decrease in the density of SERT molecules on the plasma membrane. Through the feedback effect of this proposed mechanism, plasma 5HT controls its own concentration levels by modulating the uptake properties of platelet SERT.

At diabetes-like concentration, glucose down-regulates the placental serotonin transport system in a cell-cycle-dependent manner.

Serotonin [5-hydroxytryptamine (5HT)] is a vasoconstrictor that also acts as a developmental signal early in embryogenesis. The 5HT transporter (SERT) on the membranes of the placental trophoblast cells controls 5HT levels in the maternal bloodstream to maintain stable transplacental blood flow and simultaneously provide 5HT to the embryo. The 5HT uptake rate of placental SERT is important for both the mother and the developing embryo. The impact of glucose on the placental SERT system during diabetic pregnancy is not known. The present in vitro study investigated this important issue in human placental choriocarcinoma (JAR) cells that were cultured for 24-96 h in a medium containing either 5.5 (physiologic concentration) or 25 mmol/L D-glucose (diabetic-like concentration). The 5HT uptake rates of the cultured cells were not altered at exogenous D-glucose concentrations in the range of 5.5-15 mmol/L, but were decreased significantly at a diabetic-like concentration (>or=25 mmol/L). To understand better the role of glucose on the placental 5HT system, we first characterized SERT in JAR cells at different cell-cycle phases and then determined the expression levels of SERT on the plasma membrane and in the intracellular pools of JAR cells at the late-S and G2 phases, where the uptake rates were decreased 73% under diabetic-like glucose concentrations. Finally, the importance of self-association of SERT molecules was examined. In JAR cells co-expressing Flag- and myc-tagged SERT, myc-antibody precipitated 70% of Flag-SERT, indicating that a large percentage of SERT proteins exist as oligomers in situ. Under diabetic conditions, myc-antibody no longer precipitated Flag-SERT, suggesting a disruption in the aggregation of SERT molecules. Therefore, we propose that under uncontrolled diabetic conditions, glucose down-regulates 5HT uptake rates of placental SERT by interfering with its functional expression in a cell-cycle-dependent manner.

Tracheoinnominate artery fistula following tracheostomy.

Tracheoinnominate artery fistula is a relatively rare but highly lethal complication occurring in patients with long-standing tracheostomies. Early evaluation of this problem and prompt aggressive therapy are necessary. When massive haemorrhage begins, immediate arterial compression, control of the airway and subsequent treatment of the injured artery may be lifesaving. Immediate surgical exploration through a median sternotomy is necessary to control the proximal and distal innominate artery. After the damaged artery has been excised, vascular reconstruction can be performed to preserve the connection between the proximal and distal ends of the innominate artery. A pedicled pericardial patch was successfully used for the tracheal reconstruction.