Functional Diversity of Heat-labile Toxins (LT) Produced by Enterotoxigenic Escherichia coli

Heat-labile toxins (LTs) have ADP-ribosylation activity and induce the secretory diarrhea caused by enterotoxigenic Escherichia coli (ETEC) strains in different mammalian hosts. LTs also act as adjuvants following delivery via mucosal, parenteral, or transcutaneous routes. Previously we have shown that LT produced by human-derived ETEC strains encompass a group of 16 polymorphic variants, including the reference toxin (LT1 or hLT) produced by the H10407 strain and one variant that is found mainly among bacterial strains isolated from pigs (LT4 or pLT). Herein, we show that LT4 ( with six polymorphic sites in the A (K4R, K213E, and N238D) and B (S4T, A46E, and E102K) subunits) displays differential in vitro toxicity and in vivo adjuvant activities compared with LT1. One in vitro generated LT mutant (LTK4R), in which the lysine at position 4 of the A subunit was replaced by arginine, showed most of the LT4 features with an similar to 10-fold reduction of the cytotonic effects, ADP-ribosylation activity, and accumulation of intracellular cAMP in Y1 cells. Molecular dynamic studies of the A subunit showed that the K4R replacement reduces the N-terminal region flexibility and decreases the catalytic site crevice. Noticeably, LT4 showed a stronger Th1-biased adjuvant activity with regard to LT1, particularly concerning activation of cytotoxic CD8(+) T lymphocytes when delivered via the intranasal route. Our results further emphasize the relevance of LT polymorphism among human-derived ETEC strains that may impact both the pathogenicity of the bacterial strain and the use of these toxins as potential vaccine adjuvants.

Distinctive Immunomodulatory and Inflammatory Properties of the Escherichia coli Type II Heat-Labile Enterotoxin LT-IIa and Its B Pentamer following Intradermal Administration

The type I and type II heat-labile enterotoxins (LT-I and LT-II) are strong mucosal adjuvants when they are coadministered with soluble antigens. Nonetheless, data on the parenteral adjuvant activities of LT-II are still limited. Particularly, no previous study has evaluated the adjuvant effects and induced inflammatory reactions of LT-II holotoxins or their B pentameric subunits after delivery via the intradermal (i.d.) route to mice. In the present report, the adjuvant and local skin inflammatory effects of LT-IIa and its B subunit pentamer (LT-IIaB(5)) were determined. When coadministered with ovalbumin (OVA), LT-IIa and, to a lesser extent, LT-IIaB(5) exhibited serum IgG adjuvant effects. In addition, LT-IIa but not LT-IIaB(5) induced T cell-specific anti-OVA responses, particularly in respect to induction of antigen-specific cytotoxic CD8(+) T cell responses. LT-IIa and LT-IIaB(5) induced differential tissue permeability and local inflammatory reactions after i.d. injection. Of particular interest was the reduced or complete lack of local reactions, such as edema and tissue induration, in mice i.d. inoculated with LT-IIa and LT-IIaB(5), respectively, compared with mice immunized with LT-I. In conclusion, the present results show that LT-IIa and, to a lesser extent, LT-IIaB(5) exert adjuvant effects when they are delivered via the i.d. route. In addition, the low inflammatory effects of LT-IIa and LT-IIaB(5) in comparison to those of LT-I support the usefulness of LT-IIa and LT-IIaB(5) as parenterally delivered vaccine adjuvants.

Functional and immunological characterization of a natural polymorphic variant of a heat-labile toxin (LT-I) produced by enterotoxigenic Escherichia coli (ETEC)

Heat-labile toxins (LT) encompass at least 16 natural polymorphic toxin variants expressed by wild-type enterotoxigenic Escherichia coli (ETEC) strains isolated from human beings, but only one specific form, produced by the reference ETEC H10407 strain (LT1), has been intensively studied either as a virulence-associated factor or as a mucosal/transcutaneous adjuvant. In the present study, we carried out a biological/immunological characterization of a natural LT variant (LT2) with four polymorphic sites at the A subunit (S190L, G196D, K213E, and S224T) and one at the B subunit (T75A). The results indicated that purified LT2, in comparison with LT1, displayed similar in vitro toxic activities (adenosine 3`,5`-cyclic monophosphate accumulation) on mammalian cells and in vivo immunogenicity following delivery via the oral route. Nonetheless, the LT2 variant showed increased adjuvant action to ovalbumin when delivered to mice via the transcutaneous route while antibodies raised in mice immunized with LT2 displayed enhanced affinity and neutralization activity to LT1 and LT2. Taken together, the results indicate that the two most frequent LT polymorphic forms expressed by wild ETEC strains share similar biological features, but differ with regard to their immunological properties.

Leishmania amazonensis META2 protein confers protection against heat shock and oxidative stress

The META cluster of Leishmania amazonensis contains both META1 and META2 genes, which are upregulated in metacyclic promastigotes and encode proteins containing the META domain. Previous studies defined META2 as a 48.0-kDa protein, which is conserved in other Leishmania species and in Trypanosoma brucei. In this work, we demonstrate that META2 protein expression is regulated during the Leishmania life cycle but constitutive in T. brucei. META2 protein is present in the cytoplasm and flagellum of L amazonensis promastigotes. Leishmania META2-null replacement mutants are more sensitive to oxidative stress and, upon heat shock, assume rounded morphology with shortened flagella. The increased susceptibility of null parasites to heat shock is reversed by extra-chromosomal expression of the META2 gene. Defective Leishmania promastigotes exhibit decreased ability to survive in macrophages. By contrast, META2 expression is decreased by 80% in RNAi-induced T. brucei bloodstream forms with no measurable effect on survival or resistance to heat shock. (C) 2010 Elsevier Inc. All rights reserved.

Patterns of water and heat flux across a biome gradient from tropical forest to savanna in Brazil

We investigated the seasonal patterns of water vapor and sensible heat flux along a tropical biome gradient from forest to savanna. We analyzed data from a network of flux towers in Brazil that were operated within the Large-Scale Biosphere-Atmosphere Experiment in Amazonia (LBA). These tower sites included tropical humid and semideciduous forest, transitional forest, floodplain (with physiognomies of cerrado), and cerrado sensu stricto. The mean annual sensible heat flux at all sites ranged from 20 to 38 Wm(-2), and was generally reduced in the wet season and increased in the late dry season, coincident with seasonal variations of net radiation and soil moisture. The sites were easily divisible into two functional groups based on the seasonality of evaporation: tropical forest and savanna. At sites with an annual precipitation above 1900 mm and a dry season length less than 4 months (Manaus, Santarem and Rondonia), evaporation rates increased in the dry season, coincident with increased radiation. Evaporation rates were as high as 4.0 mm d(-1) in these evergreen or semidecidous forests. In contrast, ecosystems with precipitation less than 1700 mm and a longer dry season (Mato Grosso, Tocantins and Sao Paulo) showed clear evidence of reduced evaporation in the dry season. Evaporation rates were as low as 2.5 mm d(-1) in the transitional forests and 1 mm d(-1) in the cerrado. The controls on evapotranspiration seasonality changed along the biome gradient, with evaporative demand (especially net radiation) playing a more important role in the wetter forests, and soil moisture playing a more important role in the drier savannah sites.

Pulsating strings in Lunin-Maldacena backgrounds

We consider pulsating strings in Lunin-Maldacena backgrounds, specifically in deformed Minkowski spacetime and deformed AdS(5) x S(5). We find the relation between the energy and the oscillation number of the pulsating string when the deformation is small. Since the oscillation number is an adiabatic invariant it can be used to explore the regime of highly excited string states. We then quantize the string and look for such a sector. For the deformed Minkowski background we find a precise match with the classical results if the oscillation number is quantized as an even number. For the deformed AdS(5) x S(5) we find a contribution which depends on the deformation parameter.

TL dosimetry of natural quartz sensitized by heat-treatment and high dose irradiation

The aim of this paper is to report the sensitization of the TL peak appearing at 270 degrees C in the glow curve of natural quartz by using the combined effect of heat-treatments and irradiation with high gamma doses. For this, thirty discs with 6 x 1 mm(2) were prepared from plates parallell to a rhombolledral crystal face. The specimens were separated into four lots according to its TL read out between 160 and 320 degrees C. One lot was submitted to gamma doses of Co-60 radiation starting at 2 kGy and going up until a cumulative dose of 25 kGy. The other three lots were initially heal-treated at 500, 800 and 1000 degrees C and then irradiated with a single dose of 25kGy. The TL response of each lot was determined as a function of test-doses ranging from 0.1 to 30 mGy. As a result, it was observed that heat-treatments themselves did not produce the strong peak at 270 degrees C that was observed after the administration of high gamma doses. This peak is associated with the optical absorption band appearing at 470 rim which is due to the formation of [AlO4]degrees acting as electron-hole recombination centers. The formation of the 270 degrees C peak was preliminary analyzed in relation to aluminum- and oxygen-vacancy-related centers found in crystalline quartz. (C) 2008 Elsevier Ltd. All rights reserved.

Influence of pre-heat treatment and different light-curing units on Vickers hardness of a microhybrid composite resin

The aim of this study was to evaluate the hardness of a dental composite resin submitted to temperature changes before photo-activation with two light-curing unite (LCUs). Five samples (4 mm in diameter and 2 mm in thickness) for each group were made with pre-cure temperatures of 37, 54, and 60A degrees C. The samples were photo-activated with a conventional quartz-tungsten-halogen (QTH) and blue LED LCUs during 40 s. The hardness Vickers test (VHN) was performed on the top and bottom surfaces of the samples. According to the interaction between light-curing unit and different pre-heating temperatures of composite resin, only the light-curing unit provided influences on the mean values of initial Vickers hardness. The light-curing unit based on blue LED showed hardness mean values more homogeneous between the top and bottom surfaces. The hardness mean values were not statistically significant difference for the pre-cure temperature used. According to these results, the pre-heating of the composite resin provide no influence on Vickers hardness mean values, however the blue LED showed a cure more homogeneous than QTH LCU.

Glucose-induced heat production, insulin secretion and lactate production in isolated Wistar rat pancreatic islets

Transplantation of pancreatic islets is efficient in improving the metabolic control and quality of life and in preventing severe hypoglycemia in patients with brittle type I diabetes mellitus. More accurate methods to assess islet viability would be extremely useful in designing target interventions for islet cytoprorection and in reducing the number of islets required to achieve insulin independence. Here we report on an application of calorimetry to evaluate the metabolic response of pancreatic islets to glucose stimulation. A significant increase in metabolic heat was produced by islet samples when consecutively subjected to 2.8 and 16.3 mmol L-1 glucose. Under these glucose concentrations, 1000 islets released average heat values of 9.16 +/- 0.71 mJ and 14.90 +/- 1.21 mJ over 50 min, respectively. Additionally, the glucose stimulation indexes were 1.67 +/- 0.30 for insulin. 1.72 +/- 0.13 for heat and 2.91 +/- 0.50 for lactate, raising the important possibility of substituting the secreted insulin index/ratio by the index/ratio of the heat released in the evaluation of Langerhans islets viability for transplantation. Altogether, Our results demonstrate the applicability of calorimetry to assess the quality of isolated pancreatic islets and to study vital islet functions. (c) 2008 Published by Elsevier B.V.

Stoichiometry and thermodynamics of the interaction between the C-terminus of human 90 kDa heat shock protein Hsp90 and the mitochondrial translocase of outer membrane Tom70

A large majority of the 1000-1500 proteins in the mitochondria are encoded by the nuclear genome, and therefore, they are translated in the cytosol in the form and contain signals to enable the import of proteins into the organelle. The TOM complex is the major translocase of the outer membrane responsible for preprotein translocation. It consists of a general import pore complex and two membrane import receptors, Tom20 and Tom70. Tom70 contains a characteristic TPR domain, which is a docking site for the Hsp70 and Hsp90 chaperones. These chaperones are involved in protecting cytosolic preproteins from aggregation and then in delivering them to the TOM complex. Although highly significant, many aspects of the interaction between Tom70 and Hsp90 are still uncertain. Thus, we used biophysical tools to study the interaction between the C-terminal domain of Hsp90 (C-Hsp90), which contains the EEVD motif that binds to TPR domains, and the cytosolic fragment of Tom70. The results indicate a stoichiometry of binding of one monomer of Tom70 per dimer of C-Hsp90 with a K(D) of 360 30 nM, and the stoichiometry and thermodynamic parameters obtained suggested that Tom70 presents a different mechanism of interaction with Hsp90 when compared with other TPR proteins investigated. (C) 2011 Elsevier Inc. All rights reserved.

Calucaltion of waste heat from hot rolled steel coils at SSAB and its recovery

Hot rolling process is heat input process. The heat energy in hot rolled steel coils can be utilized. At SSAB Strip Product Borlänge when the hot rolled steel coils came out of the hot rolling mill they are at the temperature range of 500°C to 800°C. Heat energy contained by the one hot rolled steel coil is about 1981Kwh whereas the total heat energy for the year 2008 is 230 GWh/year.The potential of heat is too much but the heat dissipation rate is too slow. Different factors on which heat dissipation rate depends are discussed.Three suggestions are proposed to collect the waste heat from hot rolled steel coils.The 2nd proposal in which water basin is suggested would help not only to collect the waste heat but to decrease in the cooling time.

Heat losses and thermal performance of commercial combined solar and pellet heating systems

Various pellet heating systems are marketed in Sweden, some of them in combination with a solar heating system. Several types of pellet heating units are available and can be used for a combined system. This article compares four typical combined solar and pellet heating systems: System 1 and 2 two with a pellet stove, system 3 with a store integrated pellet burner and system 4 with a pellet boiler. The lower efficiency of pellet heaters compared to oil or gas heaters increases the primary energy demand. Consequently heat losses of the various systems have been studied. The systems have been modeled in TRNSYS and simulated with parameters identified from measurements. For almost all systems the flue gas losses are the main heat losses except for system 3 where store heat losses prevail. Relevant are also the heat losses of the burner and the boiler to the ambient. Significant leakage losses are noticed for system 3 and 4. For buildings with an open internal design system 1 is the most efficient solution. Other buildings should preferably apply system 3. The right choice of the system depends also on whether the heater is placed inside or outside of the heated are. A large potential for system optimization exist for all studied systems, which when applied could alter the relative merits of the different system types.

Identifiering av parametrar till ackumulatortanken Solus1050 från consolar - en metodbeskrivning

This report describes the work done creating a computer model of a kombi tank from Consolar. The model was created with Presim/Trnsys and Fittrn and DF were used to identify the parameters. Measurements were carried out and were used to identify the values of the parameters in the model. The identifications were first done for every circuit separately. After that, all parameters are normally identified together using all the measurements. Finally the model should be compared with other measurements, preferable realistic ones. The two last steps have not yet been carried out, because of problems finding a good model for the domestic hot water circuit.The model of the domestic hot water circuit give relatively good results for low flows at 5 l/min, but is not good for higher flows. In the report suggestions for improving the model are given. However, there was not enough time to test this within the project as much time was spent trying to solve problems with the model crashing. Suggestions for improving the model for the domestic circuit are given in chapter 4.4. The improved equations that are to be used in the improved model are given by equation 4.18, 4.19 and 4.22.Also for the boiler circuit and the solar circuit there are improvements that can be done. The model presented here has a few shortcomings, but with some extra work, an improved model can be created. In the attachment (Bilaga 1) is a description of the used model and all the identified parameters.A qualitative assessment of the store was also performed based on the measurements and the modelling carried out. The following summary of this can be given: Hot Water PreparationThe principle for controlling the flow on the primary side seems to work well in order to achieve good stratification. Temperatures in the bottom of the store after a short use of hot water, at a coldwater temperature of 12°C, was around 28-30°C. This was almost independent of the temperature in the store and the DHW-flow.The measured UA-values of the heat exchangers are not very reliable, but indicates that the heat transfer rates are much better than for the Conus 500, and in the same range as for other stores tested at SERC.The function of the mixing valve is not perfect (see diagram 4.3, where Tout1 is the outlet hot water temperature, and Tdhwo and Tdhw1 is the inlet temperature to the hot and cold side of the valve respectively). The outlet temperature varies a lot with different temperatures in the storage and is going down from 61°C to 47°C before the cold port is fully closed. This gives a problem to find a suitable temperature setting and gives also a risk that the auxiliary heating is increased instead of the set temperature of the valve, when the hot water temperature is to low.Collector circuitThe UA-value of the collector heat exchanger is much higher than the value for Conus 500, and in the same range as the heat exchangers in other stores tested at SERC.Boiler circuitThe valve in the boiler circuit is used to supply water from the boiler at two different heights, depending on the temperature of the water. At temperatures from the boiler above 58.2°C, all the water is injected to the upper inlet. At temperatures below 53.9°C all the water is injected to the lower inlet. At 56°C the water flow is equally divided between the two inlets. Detailed studies of the behaviour at the upper inlet shows that better accuracy of the model would have been achieved using three double ports in the model instead of two. The shape of the upper inlet makes turbulence, that could be modelled using two different inlets. Heat lossesThe heat losses per m3 are much smaller for the Solus 1050, than for the Conus 500 Storage. However, they are higher than those for some good stores tested at SERC. The pipes that are penetrating the insulation give air leakage and cold bridges, which could be a major part of the losses from the storage. The identified losses from the bottom of the storage are exceptionally high, but have less importance for the heat losses, due to the lower temperatures in the bottom. High losses from the bottom can be caused by air leakage through the insulation at the pipe connections of the storage.