Insights into the biochemical strategies of adaptation to heat stress in the hyperthermophilic archaeon Pyrococcus furiosus

Organisms that thrive optimally at temperatures above 80°C are called hyperthermophiles. These prokaryotes have been isolated from a variety of hot environments, such as marine geothermal areas, hence they are usually slightly halophilic. Like other halophiles, marine hyperthermophiles have to cope with fluctuations in the salinity of the external medium and generally use low-molecular mass organic compounds to adjust cell turgor pressure. These compounds can accumulate to high levels without interfering with cell metabolism, thereby deserving the designation of compatible solutes. Curiously, the accumulation of compatible solutes also occurs in response to supraoptimal temperatures.(...)

Exploring the relationship between toxin and spore production in the human enteric pathogen Clostridium difficile

RESUMO: Clostridium difficile é presentemente a principal causa de doença gastrointestinal associada à utilização de antibióticos em adultos. C. difficile é uma bactéria Gram-positiva, obrigatoriamente anaeróbica, capaz de formar endósporos. Tem-se verificado um aumento dos casos de doença associada a C. difficile com sintomas mais severos, elevadas taxas de morbilidade, mortalidade e recorrência, em parte, devido à emergência de estirpes mais virulentas, mas também devido à má gestão do uso de antibióticos. C. difficile produz duas toxinas, TcdA e TcdB, que são os principais fatores de virulência e responsáveis pelos sintomas da doença. Estas são codificadas a partir do Locus de Patogenicidade (PaLoc) que codifica ainda para um regulador positivo, TcdR, uma holina, TcdE, e um regulador negativo, TcdC. Os esporos resistentes ao oxigénio são essenciais para a transmissão do organismo e recorrência da doença. A expressão dos genes do PaLoc ocorre em células vegetativas, no final da fase de crescimento exponencial, e em células em esporulação. Neste trabalho construímos dois mutantes de eliminação em fase dos genes tcdR e tcdE. Mostrámos que a auto-regulação do gene tcdR não é significativa. No entanto, tcdR é sempre necessário para a expressão dos genes presentes no PaLoc. Trabalho anterior mostrou que, com a exceção de tcdC, os demais genes do PaLoc são expressos no pré-esporo. Mostrámos aqui que TcdA é detectada à superfície do esporo maduro e que a eliminação do tcdE não influencia a acumulação de TcdA no meio de cultura ou em associação às células ou ao esporo. Estas observações têm consequências para o nosso entendimento do processo infecioso: sugeremque o esporo possa ser também um veículo para a entrega da toxina nos estágios iniciais da infecção, que TcdA possa ser libertada durante a germinação do esporo, e que o esporo possa utilizar o mesmo receptor reconhecido por TcdA para a ligação à mucosa do cólon.---------------------------ABSTRACT: Clostridium difficile is currently the major cause of antibiotic-associated gastrointestinal diseases in adults. This is a Gram-positive bacterium, endospore-forming and an obligate anaerobe that colonizes the gastrointestinal tract. Recent years have seen a rise in C. difficile associated disease (CDAD) cases, associated with more severe disease symptoms, higher rates of morbidity, mortality and recurrence, which were mostly caused due to the emergence of “hypervirulent” strains but also due to changing patterns of antibiotics use. C. difficile produces two potent toxins, TcdA and TcdB, which are the main virulence factors and the responsible for the disease symptoms. These are codified from a Pathogenicity Locus (PaLoc), composed also by the positive regulator, TcdR, the holin-like protein, TcdE, and a negative regulator, TcdC. Besides the toxins, the oxygen-resistant spores are also essential for transmission of the organism through diarrhea; moreover, spores can accumulate in the environment or in the host, which will cause disease recurrence. The expression of the PaLoc genes occurs in vegetative cells, at the end of the exponential growth phase, and in sporulating cells. In this work, we constructed two in-frame deletion mutants of tcdR and tcdE. We showed that the positive auto regulation of tcdR is not significant. However, tcdR is always necessary for the expression of the PaLoc genes. A previous work showed that, except tcdC, all the PaLoc genes are expressed in the forespore. Here, we detected TcdA at the spore surface. Furthermore, we showed that the in-frame deletion of tcdE does not affect the accumulation of TcdA in the culture medium or in association with cells or spores. This data was important for us to conclude about the infeccious process: it suggests that the spore may be the vehicle for the delivery of TcdA in early stages of infection, that TcdA may be released during spores germination and that this spore may use the same receptor recognized by TcdA to bind to the colonic mucosa.

Microcalorimetry Assay for Rapid Detection of Voriconazole Resistance in Aspergillus fumigatus.

We describe a calorimetric assay for detection of voriconazole-resistant Aspergillus fumigatus within 8 h. Among 27 genetically distinct strains, all 21 resistant and all 6 susceptible strains were correctly identified by measurement of fungal heat production in the presence of voriconazole. This proof-of-concept study demonstrates the potential of microcalorimetry for rapid detection of azole resistance in A. fumigatus.

Seasonal changes in behavioural and thermoregulatory responses to hypoxia in the Eastern Chipmunk (Tamias striatus) /

Mammalian heterotherms, such as hibemators, are known to be more tolerant of low oxygen tensions than their homeothermic counterparts. It has been suggested that this relative hypoxia tolerance is related to their ability to deal with dramatic changes in body temperature during entry to and arousal from torpor. However, hibemators demonstrate dramatic seasonality in both daily heterothermy and overall torpor expression. It was of interest to test if seasonal comparisons of normothermic individuals within a single species with the capacity to hibernate produce changes in the response to hypoxia that would reflect a seasonal change in tolerance to low oxygen. In particular, the species studied, the Eastern chipmunk {Tamias striatus), is known to enter into torpor exclusively in the winter. To test for seasonal differences in the metabolic and thermoregulatory responses to hypoxia, flow-through respirometry was used to compare metabolic rate, minimum thermal conductance, body temperature, and a thermal gradient used to assess selected ambient temperature in response to hypoxia in both summer and winter acclimated animals. Although the animals periodically expressed torpor throughout the winter, no differences between season in resting metabolic rate, body temperature or minimum thermal conductance were observed in normoxia. The metabolic trials indicated that chipmunks are less responsive to hypoxia in the winter than they are in the summer. Although body temperature dropped in response to hypoxia in both seasons, the decrease was less in the winter, and there was no corresponding decrease in metabolic rate. Providing the animals with a choice of ambient temperatures in hypoxia resulted in a blunting of the drop in body temperature in both seasons, suggesting that the reported fall in body temperature set point in hypoxia is not fully manifested in the behavioural pathways responsible for thermoregulation in chipmunks. Instead, body temperature in hypoxia appears to be highly dependent on ambient temperature and oxygen concentration. The results of this study suggest that the season in which the responses to hypoxia are measured is important, especially in a heterotherm where seasonality can affect the degree to 1 which the animal is tolerant of hypoxia. Winter-acclimated chipmunks appear more capable of defending metabolic heat production in hypoxia, a response consistent with the increased thermogenic capacity observed in animals that must periodically enter and arouse from torpor during hibernation.

Mechanisms of endothelin-1 induced reactive oxygen species production in vascular adventitial fibroblasts

With the relationship between endothelin-1 (ET-1) stimulation and reactive oxygen species (ROS) production unknown in adventitial fibroblasts, I examined the ROS response to ET-1 and angiotensin (Ang II). ET-1 -induced ROS peaked following 4 hrs of ET-1 stimulation and was inhibited by an ETA receptor antagonist (BQ 123, 1 uM) an extracellular signal-regulated kinase (ERK) 1/2 inhibitor (PD98059, 10 uM), and by both a specific, apocynin (10 uM), and non-specific, diphenyleneiodonium (10 uM), NAD(P)H oxidase inhibitor. NOX2 knockout fibroblasts did not produce an ET-1 induced change in ROS levels. Ang II treatment increased ROS levels in a biphasic manner, with the second peak occurring 6 hrs following stimulation. The secondary phase of Ang II induced ROS was inhibited by an ATi receptor antagonist, Losartan (100 uM) and BQ 123. In conclusion, ET-1 induces ROS production primarily through an ETA-ERKl/2 NOX2 pathway, additionally, Ang II-induced ROS production also involves an ETa pathway.

Estimation of Steel Exports & Imports to analyze the Impact of Free Trade on Production in the Steel Industry.

Rapport de recherche

An Economic Analysis of Wind Energy Production in the Remote Areas of Quebec.

Rapport de recherche

Photothermal deflection measurement on heat transport in GaAs epitaxial layers

In this paper, we report the in-plane and cross-plane measurements of the thermal diffusivity of double epitaxial layers of n-type GaAs doped with various concentrations of Si and a p-type Be-doped GaAs layer grown on a GaAs substrate by the molecular beam epitaxial method, using the laser-induced nondestructive photothermal deflection technique. The thermal diffusivity value is evaluated from the slope of the graph of the phase of the photothermal deflection signal as a function of pump-probe offset. Analysis of the data shows that the cross-plane thermal diffusivity is less than that of the in-plane thermal diffusivity. It is also seen that the doping concentration has a great influence on the thermal diffusivity value. Measurement of p-type Be-doped samples shows that the nature of the dopant also influences the effective thermal diffusivity value. The results are interpreted in terms of a phonon-assisted heat transfer mechanism and the various scattering process involved in the propagation of phonons.

Photothermal deflection measurement on heat transport in GaAs epitaxial layers

In this paper, we report the in-plane and cross-plane measurements of the thermal diffusivity of double epitaxial layers of n-type GaAs doped with various concentrations of Si and a p-type Be-doped GaAs layer grown on a GaAs substrate by the molecular beam epitaxial method, using the laser-induced nondestructive photothermal deflection technique. The thermal diffusivity value is evaluated from the slope of the graph of the phase of the photothermal deflection signal as a function of pump-probe offset. Analysis of the data shows that the cross-plane thermal diffusivity is less than that of the in-plane thermal diffusivity. It is also seen that the doping concentration has a great influence on the thermal diffusivity value. Measurement of p-type Be-doped samples shows that the nature of the dopant also influences the effective thermal diffusivity value. The results are interpreted in terms of a phonon-assisted heat transfer mechanism and the various scattering process involved in the propagation of phonons

Photothermal deflection measurement on heat transport in GaAs epitaxial layers

In this paper, we report the in-plane and cross-plane measurements of the thermal diffusivity of double epitaxial layers of n-type GaAs doped with various concentrations of Si and a p-type Be-doped GaAs layer grown on a GaAs substrate by the molecular beam epitaxial method, using the laser-induced nondestructive photothermal deflection technique. The thermal diffusivity value is evaluated from the slope of the graph of the phase of the photothermal deflection signal as a function of pump-probe offset. Analysis of the data shows that the cross-plane thermal diffusivity is less than that of the in-plane thermal diffusivity. It is also seen that the doping concentration has a great influence on the thermal diffusivity value. Measurement of p-type Be-doped samples shows that the nature of the dopant also influences the effective thermal diffusivity value. The results are interpreted in terms of a phonon-assisted heat transfer mechanism and the various scattering process involved in the propagation of phonons

Photothermal deflection studies on heat transport in intrinsic and extrinsic InP

Laser induced transverse photothermal deflection technique has been employed to determine the thermal parameters of InP doped with Sn, S and Fe as well as intrinsic InP. The thermal diffusivity values of these various samples are evaluated from the slope of the curve plotted between the phase of photothermal deflection signal and pump-probe offset. Analysis of the data shows that heat transport and hence the thermal diffusivity value, is greatly affected by the introduction of dopant. It is also seen that the direction of heat flow with respect to the plane of cleavage of semiconductor wafers influences the thermal diffusivity value. The results are explained in terms of dominating phonon assisted heat transfer mechanism in semiconductors.

Studies on Heat Diffusion in Selected Conducting Polymers using Probe Beam Deflection and Photoacoustic Techniques

In the present study, radio frequency plasma polymerization technique is used to prepare thin films of polyaniline, polypyrrole, poly N-methyl pyrrole and polythiophene. The thermal characterization of these films is carried out using transverse probe beam deflection method. Electrical conductivity and band gaps are also determined. The effect of iodine doping on electrical conductivity and the rate of heat diffusion is explored.Bulk samples of poyaniline and polypyrrole in powder form are synthesized by chemical route. Open photoacoustic cell configuration is employed for the thermal characterization of these samples. The effect of acid doping on heat diffusion in these bulk samples of polyaniline is also investigated. The variation of electrical conductivity of doped polyaniline and polypyrrole with temperature is also studied for drawing conclusion on the nature of conduction in these samples. In order to improve the processability of polyaniline and polypyrrole, these polymers are incorporated into a host matrix of poly vinyl chloride. Measurements of thermal diffusivity and electrical conductivity of these samples are carried out to investigate the variation of these quantities as a function of the content of polyvinyl chloride.