Disruption of the gene for hsp30, an alpha-crystallin-related heat shock protein of Neurospora crassa, causes defects in thermotolerance.

The alpha-crystallin-related heat shock proteins are produced by all eukaryotes, but the role of these proteins in thermoprotection remains unclear. To investigate the function of one of these proteins, we disrupted expression of the single-copy hsp30 gene of Neurospora crassa, using repeat-induced point mutagenesis, and we generated and characterized mutant strains that were deficient in hsp30 synthesis. These strains could grow at high temperature and they acquired thermotolerance from a heat shock. However, the hsp30-defective strains proved to be extremely sensitive to the combined stresses of high temperature and carbohydrate limitation, enforced by the addition of a nonmetabolizable glucose analogue. Under these conditions, their survival was reduced by 90% compared with wild-type cells. This sensitive phenotype was reversed by reintroduction of a functional hsp30 gene into the mutant strains. The mutant cells contained mitochondria from which a 22-kDa protein was readily extracted with detergents, in contrast to its retention by the mitochondria of wild-type cells. Antibodies against hsp30 coimmunoprecipitated a protein also of approximately 22 kDa from wild-type cells. Results of this study suggest that hsp30 may be important for efficient carbohydrate utilization during high temperature stress and that it may interact with other mitochondrial membrane proteins and function as a protein chaperone.

Heat shock protein hsp90 regulates dioxin receptor function in vivo.

The dioxin (aryl hydrocarbon) receptor is a ligand-dependent basic helix-loop-helix (bHLH) factor that binds to xenobiotic response elements of target promoters upon heterodimerization with the bHLH partner factor Arnt. Here we have replaced the bHLH motif of the dioxin receptor with a heterologous DNA-binding domain to create fusion proteins that mediate ligand-dependent transcriptional enhancement in yeast (Saccharomyces cerevisiae). Previously, our experiments indicated that the ligand-free dioxin receptor is stably associated with the 90-kDa heat shock protein, hsp90. To investigate the role of hsp90 in dioxin signaling we have studied receptor function in a yeast strain where hsp90 expression can be down-regulated to about 5% relative to wild-type levels. At low levels of hsp90, ligand-dependent activation of the chimeric dioxin receptor construct was almost completely inhibited, whereas the activity of a similar chimeric construct containing the structurally related Arnt factor was not affected. Moreover, a chimeric dioxin receptor construct lacking the central ligand- and hsp90-binding region of the receptor showed constitutive transcriptional activity in yeast that was not impaired upon down-regulation of hsp90 expression levels. Thus, these data suggest that hsp90 is a critical determinant of conditional regulation of dioxin receptor function in vivo via the ligand-binding domain.

Degradation of sigma 32, the heat shock regulator in Escherichia coli, is governed by HflB.

The heat shock response in Escherichia coli is governed by the concentration of the highly unstable sigma factor sigma 32. The essential protein HflB (FtsH), known to control proteolysis of the phage lambda cII protein, also governs sigma 32 degradation: an HflB-depleted strain accumulated sigma 32 and induced the heat shock response, and the half-life of sigma 32 increased by a factor up to 12 in mutants with reduced HflB function and decreased by a factor of 1.8 in a strain overexpressing HflB. The hflB gene is in the ftsJ-hflB operon, one promoter of which is positively regulated by heat shock and sigma 32. The lambda cIII protein, which stabilizes sigma 32 and lambda cII, appears to inhibit the HflB-governed protease. The E. coli HflB protein controls the stability of two master regulators, lambda cII and sigma 32, responsible for the lysis-lysogeny decision of phage lambda and the heat shock response of the host.

Effect of Si additions and heat treatment on the mechanical behaviour of an Al-5Mg casting alloy

Adding 1%Si to binary Al-5Mg alloy slightly increases the yield stress in comparison with the Si free alloy but dramatically reduces the ductility and tensile strength due to the formation of brittle eutectic Mg2Si and pi-Al8FeMg3Si6 particles. Adding 3%Si slightly reduces the yield stress, presumably due to some of the Mg being tied up in the Mg2Si, and further reduces the ductility due to the increased volume fraction of intermetallics. Solution heat treatment at 436degreesC decreases the yield stress of both Si containing alloys, and slightly increases the ductility in the alloy with 3%Si. Subsequent ageing at 180degreesC has no further effects on the strength or ductility. The loss in strength of the heat treated alloys seems to be due to overageing Of Mg2Si precipitates dispersed in the bulk of the alloy. (C) 2004 W. S. Maney Son Ltd.

Large-eddy simulation of heat transfer downstream of a backward-facing step

Large-eddy simulation is used to predict heat transfer in the separated and reattached flow regions downstream of a backward-facing step. Simulations were carried out at a Reynolds number of 28 000 (based on the step height and the upstream centreline velocity) with a channel expansion ratio of 1.25. The Prandtl number was 0.71. Two subgrid-scale models were tested, namely the dynamic eddy-viscosity, eddy-diffusivity model and the dynamic mixed model. Both models showed good overall agreement with available experimental data. The simulations indicated that the peak in heat-transfer coefficient occurs slightly upstream of the mean reattachment location, in agreement with experimental data. The results of these simulations have been analysed to discover the mechanisms that cause this phenomenon. The peak in heat-transfer coefficient shows a direct correlation with the peak in wall shear-stress fluctuations. It is conjectured that the peak in these fluctuations is caused by an impingement mechanism, in which large eddies, originating in the shear layer, impact the wall just upstream of the mean reattachment location. These eddies cause a 'downwash', which increases the local heat-transfer coefficient by bringing cold fluid from above the shear layer towards the wall.

Experimental and analytical study of the effect of contact angle on liquid convective heat transfer in microchannels

In this paper we examine the effect of contact angle (or surface wettability) on the convective heat transfer coefficient in microchannels. Slip flow, where the fluid velocity at the wall is non-zero, is most likely to occur in microchannels due to its dependence on shear rate or wall shear stress. We show analytically that for a constant pressure drop, the presence of slip increases the Nusselt number. In a microchannel heat exchanger we modified the surface wettability from a contact angle of 20 degrees-120 degrees using thin film coating technology. Apparent slip flow is implied from pressure and flow rate measurements with a departure from classical laminar friction coefficients above a critical shear rate of approximately 10,000 s(-1). The magnitude of this departure is dependant on the contact angle with higher contact angles surfaces exhibiting larger pressure drop decreases. Similarly, the non-dimensional heat flux is found to decrease relative to laminar non-slip theory, and this decrease is also a function of the contact angle. Depending on the contact angle and the wall shear rate, variations in the heat transfer rate exceeding 10% can be expected. Thus the contact angle is an important consideration in the design of micro, and even more so, nano heat exchangers. (c) 2006 Elsevier Ltd. All rights reserved.

PerR controls Mn-dependent resistance to oxidative stress in Neisseria gonorrhoeae

In previous studies it has been established that resistance to superoxide by Neisseria gonorrhoeae is dependent on the accumulation of Mn(II) ions involving the ABC transporter, MntABC. A mutant strain lacking the periplasmic binding protein component (MntC) of this transport system is hypersensitive to killing by superoxide anion. In this study the mntC mutant was found to be more sensitive to H2O2 killing than the wild-type. Analysis of regulation of MntC expression revealed that it was de-repressed under low Mn(II) conditions. The N. gonorrhoeae mntABC locus lacks the mntR repressor typically found associated with this locus in other organisms. A search for a candidate regulator of mntABC expression revealed a homologue of PerR, a Mn-dependent peroxide-responsive regulator found in Gram-positive organisms. A perR mutant expressed more MntC protein than wild-type, and expression was independent of Mn(II), consistent with a role for PerR as a repressor of mntABC expression. The PerR regulon of N. gonorrhoeae was defined by microarray analysis and includes ribosomal proteins, TonB-dependent receptors and an alcohol dehydrogenase. Both the mntC and perR mutants had reduced intracellular survival in a human cervical epithelial cell model.

Short-term hyperthermic treatment of Penaeus monodon increases expression of heat shock protein 70 (HSP70) and reduces replication of gill associated virus (GAV)

Disease is the result of interactions amongst pathogens, the environment and host organisms. To investigate the effect of stress on Penaeus monodon, juvenile shrimp were given short term exposure to hypoxic, hyperthermic and osmotic stress twice over a 1-week period and estimates of total haemocyte count (THC), heat shock protein (HSP) 70 expression and load of gill associated virus (GAV) were determined at different time points. While no significant differences were observed in survival and THC between stressed and control shrimp (P>0.05), HSP 70 expression and GAV load changed significantly (P

Cleavage initiation in the intercritically reheated coarse-grained heat-affected zone:Part I. Fractographic evidence

Tensile, crack opening displacement (COD), blunt notch, and Charpy impact tests were used to investigate cleavage initiation in the intercritically reheated coarse-grained heat-affected zone (IC CG HAZ) of three steels. The steels were chosen to provide different distributions and morphologies of MA (high-carbon martensite with some retained austenite) particles within the IC CG HAZ structure. Observation of minimum impact toughness values for the IC CG HAZ was found to be associated with a particular microstructure containing a near-connected grain boundary network of blocky MA particles, the MA particles being significantly harder than the internal grain microstructure. The initiation mechanism for this structure was determined to be from a combination of an overlap of residual transformational induced stress fields, due to the formation of the MA particles, between two closely spaced particles and stress concentration effects resulting from debonding of the particles. © 1994 The Minerals, Metals and Materials Society, and ASM International.

Repeated evolution of heat responsiveness among Brassicaceae COPIA transposable elements

Eukaryotic genomes contain repetitive DNA sequences. This includes simple repeats and more complex transposable elements (TEs). Many TEs reach high copy numbers in the host genome, owing to their amplification abilities by specific mechanisms. There is growing evidence that TEs contribute to gene transcriptional regulation. However, excess of TE activity may lead to reduced genome stability. Therefore, TEs are suppressed by the transcriptional gene silencing machinery via specific chromatin modifications. In contrary, effectiveness of the epigenetic silencing mechanisms imposes risk for TE survival in the host genome. Therefore, TEs may have evolved specific strategies for bypassing epigenetic control and allowing the emergence of new TE copies. Recent studies suggested that the epigenetic silencing can be, at least transiently, attenuated by heat stress in A. thaliana. Heat stress induced strong transcriptional activation of COPIA78 family LTR-retrotransposons named ONSEN, and even their transposition in mutants deficient in siRNA-biogenesis. ONSEN transcriptional activation was facilitated by the presence of heat responsive elements (HREs) within the long terminal repeats, which serve as a binding platform for the HEAT SHOCK FACTORs (HSFs). This thesis focused on the evolution of ONSEN heat responsiveness in Brassicaceae. By using whole-transcriptome sequencing approach, multiple Arabidopsis lyrata ONSENs with conserved heat response were found and together with ONSENs from other Brassicaceae were used to reconstruct the evolution of ONSEN HREs. This indicated ancestral situation with two, in palindrome organized, HSF binding motifs. In the genera Arabidopsis and Ballantinia, a local duplication of this locus increased number of HSF binding motifs to four, forming a high-efficiency HRE. In addition, whole transcriptome analysis revealed novel heat-responsive TE families COPIA20, COPIA37 and HATE. Notably, HATE represents so far unknown COPIA family which occurs in several Brassicaceae species but is absent in A. thaliana. Putative HREs were identified within the LTRs of COPIA20, COPIA37 and HATE of A. lyrata, and could be preliminarily validated by transcriptional analysis upon heat induction in subsequent survey of Brassicaeae species. Subsequent phylogenetic analysis indicated a repeated evolution of heat responsiveness within Brassicaceae COPIA LTR-retrotransposons. This indicates that acquisition of heat responsiveness may represent a successful strategy for survival of TEs within the host genome.

Characterization of heat waves affecting mortality rates of broilers between 29 days and market age

Climate may affect broiler production, especially where there are heat waves, which may cause high mortality rates due to the heat stress. Heat wave prediction and characterization may allow early mitigation actions to be taken. Data Mining is one of the tools used for such a characterization, particularly when a large number of variables is involved. The objective of this study was to classify heat waves that promote broiler chicken mortality in poultry houses equipped with minimal environmental control. A single day of heat, a heat-shock day, is capable of producing high broiler mortality. In poultry houses equipped with fans and evaporative cooling, the characterization of heat waves affecting broiler mortality between 29 days of age and market age presented 89.34% Model Accuracy and 0.73 Class Precision for high mortality. There was no influence on high mortality (HM) of birds between 29 and 31 days of age. Maximum temperature humidity index (THI) above 30.6 ºC was the main characteristic of days when there was a heat wave, causing high mortality in broilers older than 31 days. The high mortality of broilers between 31 and 40 days of age occurred when maximum THI was above 30.6 ºC and maximum temperature of the day was above 34.4 ºC. There were two main causes of high mortality of broilers older than 40 days: 1) maximum THI above 30.6 ºC and minimum THI equal or lower than 15.5 ºC; 2) maximum THI above 30.6 ºC, minimum THI lower than 15.5 ºC, and the time of maximum temperature later than 15:00h. The heat wave influence on broiler mortality lasted an average of 2.7 days.

Tonsils of the soft palate do not mediate the response of pigs to oral vaccination with heat-inactivated Mycobacterium bovis

Mycobacterium bovis causes animal tuberculosis (TB) in cattle, humans, and other mammalian species, including pigs. The goal of this study was to experimentally assess the responses of pigs with and without a history of tonsillectomy to oral vaccination with heat-inactivated M. bovis and challenge with a virulent M. bovis field strain, to compare pig and wild boar responses using the same vaccination model as previously used in the Eurasian wild boar (Sus scrofa), to evaluate the use of several enzyme-linked immunosorbent assays (ELISAs) and lateral flow tests for in vivo TB diagnosis in pigs, and to verify if these tests are influenced by oral vaccination with inactivated M. bovis. At necropsy, the lesion and culture scores were 20% to 43% higher in the controls than those in the vaccinated pigs. Massive M. bovis growth from thoracic tissue samples was observed in 4 out of 9 controls but in none of the 10 vaccinated pigs. No effect of the presence or absence of tonsils was observed on these scores, suggesting that tonsils are not involved in the protective response to this vaccine in pigs. The serum antibody levels increased significantly only after challenge. At necropsy, the estimated sensitivities of the ELISAs and dual path platform (DPP) assays ranged from 89% to 94%. In the oral mucosa, no differences in gene expression were observed in the control group between the pigs with and without tonsils. In the vaccinated group, the mRNA levels for chemokine (C-C motif) receptor 7 (CCR7), interferon beta (IFN-β), and methylmalonyl coenzyme A mutase (MUT) were higher in pigs with tonsils. Complement component 3 mRNA levels in peripheral blood mononuclear cells (PBMC) increased with vaccination and decreased after M. bovis challenge. This information is relevant for pig production in regions that are endemic for M. bovis and for TB vaccine research.

Protection against Tuberculosis in Eurasian Wild Boar Vaccinated with Heat-Inactivated Mycobacterium bovis

Tuberculosis (TB) caused by Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex continues to affect humans and animals worldwide and its control requires vaccination of wildlife reservoir species such as Eurasian wild boar (Sus scrofa). Vaccination efforts for TB control in wildlife have been based primarily on oral live BCG formulations. However, this is the first report of the use of oral inactivated vaccines for controlling TB in wildlife. In this study, four groups of 5 wild boar each were vaccinated with inactivated M. bovis by the oral and intramuscular routes, vaccinated with oral BCG or left unvaccinated as controls. All groups were later challenged with a field strain of M. bovis. The results of the IFN-gamma response, serum antibody levels, M. bovis culture, TB lesion scores, and the expression of C3 and MUT genes were compared between these four groups. The results suggested that vaccination with heat-inactivated M. bovis or BCG protect wild boar from TB. These results also encouraged testing combinations of BCG and inactivated M. bovis to vaccinate wild boar against TB. Vaccine formulations using heat-inactivated M. bovis for TB control in wildlife would have the advantage of being environmentally safe and more stable under field conditions when compared to live BCG vaccines. The antibody response and MUT expression levels can help differentiating between vaccinated and infected wild boar and as correlates of protective response in vaccinated animals. These results suggest that vaccine studies in free-living wild boar are now possible to reveal the full potential of protecting against TB using oral M. bovis inactivated and BCG vaccines

HEAT MANAGEMENT IN THE PUMPING INFRASTRUCTURE AREA OF AN ESTERHAZY MINE

Heat management in mines is a growing issue as mines expand physically in size and depth and as the infrastructure grows that is required to maintain them. Heat management is a concern as it relates to the health and safety of the workers as set by the regulations of governing bodies as well as the heat sensitive equipment that may be found throughout the mine workings. In order to reduce the exposure of working in hot environments there are engineering and management systems that can monitor and control the environmental conditions within the mine. The successful implementation of these methods can manage the downtime caused by heat stress environments, which can increase overall production. This thesis introduces an approach to monitoring and data based heat management. A case study is presented with an in depth approach to data collection. Data was collected for a period of up to and over one year. Continuous monitoring was conducted by equipment that was developed both commercially and within the mine site. The monitoring instrumentation was used to assess the environmental conditions found within the study area. Analysis of the data allowed for an engineering assessment of viable options in order to control and manage the environment heat stress. An option is developed and presented which allows for the greatest impact on the heat stress conditions within the case study area and is economically viable for the mine site.