Physical interaction between bacterial heat shock protein (Hsp) 90 and Hsp70 chaperones mediates their cooperative action to refold denatured proteins.

In eukaryotes, heat shock protein 90 (Hsp90) is an essential ATP-dependent molecular chaperone that associates with numerous client proteins. HtpG, a prokaryotic homolog of Hsp90, is essential for thermotolerance in cyanobacteria, and in vitro it suppresses the aggregation of denatured proteins efficiently. Understanding how the non-native client proteins bound to HtpG refold is of central importance to comprehend the essential role of HtpG under stress. Here, we demonstrate by yeast two-hybrid method, immunoprecipitation assays, and surface plasmon resonance techniques that HtpG physically interacts with DnaJ2 and DnaK2. DnaJ2, which belongs to the type II J-protein family, bound DnaK2 or HtpG with submicromolar affinity, and HtpG bound DnaK2 with micromolar affinity. Not only DnaJ2 but also HtpG enhanced the ATP hydrolysis by DnaK2. Although assisted by the DnaK2 chaperone system, HtpG enhanced native refolding of urea-denatured lactate dehydrogenase and heat-denatured glucose-6-phosphate dehydrogenase. HtpG did not substitute for DnaJ2 or GrpE in the DnaK2-assisted refolding of the denatured substrates. The heat-denatured malate dehydrogenase that did not refold by the assistance of the DnaK2 chaperone system alone was trapped by HtpG first and then transferred to DnaK2 where it refolded. Dissociation of substrates from HtpG was either ATP-dependent or -independent depending on the substrate, indicating the presence of two mechanisms of cooperative action between the HtpG and the DnaK2 chaperone system.

Conception rate of artificially inseminated Holstein cows affected by cloudy vaginal mucus, under intense heat conditions

The objective of this work was to obtain prevalence estimates of cloudy vaginal mucus in artificially inseminated Holstein cows raised under intense heat, in order to assess the effect of meteorological conditions on its occurrence during estrus and to determine its effect on conception rate. In a first study, an association was established between the occurrence of cloudy vaginal mucus during estrus and the conception rate of inseminated cows (18,620 services), raised under intense heat (mean annual temperature of 22°C), at highly technified farms, in the arid region of northern Mexico. In a second study, data from these large dairy operations were used to assess the effect of meteorological conditions throughout the year on the occurrence of cloudy vaginal mucus during artificial insemination (76,899 estruses). The overall rate of estruses with cloudy vaginal mucus was 21.4% (16,470/76,899; 95% confidence interval = 21.1-21.7%). The conception rate of cows with clean vaginal mucus was higher than that of cows with abnormal mucus (30.6 vs. 22%). Prevalence of estruses with cloudy vaginal mucus was strongly dependent on high ambient temperature and markedly higher in May and June. Acceptable conception rates in high milk-yielding Holstein cows can only be obtained with cows showing clear and translucid mucus at artificial insemination.

Simulation of Boundary Layers and Heat Transfer in the Entrance Region of Pipes

The study of fluid flow in pipes is one of the main topic of interest for engineers in industries. In this thesis, an effort is made to study the boundary layers formed near the wall of the pipe and how it behaves as a resistance to heat transfer. Before few decades, the scientists used to derive the analytical and empirical results by hand as there were limited means available to solve the complex fluid flow phenomena. Due to the increase in technology, now it has been practically possible to understand and analyze the actual fluid flow in any type of geometry. Several methodologies have been used in the past to analyze the boundary layer equations and to derive the expression for heat transfer. An integral relation approach is used for the analytical solution of the boundary layer equations and is compared with the FLUENT simulations for the laminar case. Law of the wall approach is used to derive the empirical correlation between dimensionless numbers and is then compared with the results from FLUENT for the turbulent case. In this thesis, different approaches like analytical, empirical and numerical are compared for the same set of fluid flow equations.

Flow phenomena, heat and mass transfer in microchannel reactors

The studies of flow phenomena, heat and mass transfer in microchannel reactors are beneficial to estimate and evaluate the ability of microchannel reactors to be operated for a given process reaction such as Fischer-Tropsch synthesis. The flow phenomena, for example, the flow regimes and flow patterns in microchannel reactors for both single phase and multiphase flow are affected by the configuration of the flow channel. The reviews of the previous works about the analysis of related parameters that affect the flow phenomena are shown in this report. In order to predict the phenomena of Fischer-Tropsch synthesis in microchannel reactors, the 3-dimensional computational fluid dynamic simulation with commercial software package FLUENT was done to study the flow phenomena and heat transfer for gas phase Fischer-Tropsch products flow in rectangular microchannel with hydraulic diameter 500 ¿m and length 15 cm. Numerical solution with slip boundary condition was used in the simulation and the flowphenomena and heat transfer were determined.

Chemical chaperones regulate molecular chaperones in vitro and in cells under combined salt and heat stresses.

Salt and heat stresses, which are often combined in nature, induce complementing defense mechanisms. Organisms adapt to high external salinity by accumulating small organic compounds known as osmolytes, which equilibrate cellular osmotic pressure. Osmolytes can also act as "chemical chaperones" by increasing the stability of native proteins and assisting refolding of unfolded polypeptides. Adaptation to heat stress depends on the expression of heat-shock proteins, many of which are molecular chaperones, that prevent protein aggregation, disassemble protein aggregates, and assist protein refolding. We show here that Escherichia coli cells preadapted to high salinity contain increased levels of glycine betaine that prevent protein aggregation under thermal stress. After heat shock, the aggregated proteins, which escaped protection, were disaggregated in salt-adapted cells as efficiently as in low salt. Here we address the effects of four common osmolytes on chaperone activity in vitro. Systematic dose responses of glycine betaine, glycerol, proline, and trehalose revealed a regulatory effect on the folding activities of individual and combinations of chaperones GroEL, DnaK, and ClpB. With the exception of trehalose, low physiological concentrations of proline, glycerol, and especially glycine betaine activated the molecular chaperones, likely by assisting local folding in chaperone-bound polypeptides and stabilizing the native end product of the reaction. High osmolyte concentrations, especially trehalose, strongly inhibited DnaK-dependent chaperone networks, such as DnaK+GroEL and DnaK+ClpB, likely because high viscosity affects dynamic interactions between chaperones and folding substrates and stabilizes protein aggregates. Thus, during combined salt and heat stresses, cells can specifically control protein stability and chaperone-mediated disaggregation and refolding by modulating the intracellular levels of different osmolytes.

Heat Transfer Analysis of the EPR Core Catcher Test Facility Volley

Uusi EPR-reaktorikonsepti on suunniteltu selviytymään tapauksista, joissa reaktorinsydän sulaa ja sula puhkaisee paineastian. Suojarakennuksen sisälle on suunniteltu alue, jolle sula passiivisesti kerätään, pidätetään ja jäähdytetään. Alueelle laaditaan valurautaelementeistä ns.sydänsieppari, joka tulvitetaan vedellä. Sydänsulan tuottama jälkilämpö siirtyyveteen, mistä se poistetaan suojarakennuksen jälkilämmönpoistojärjestelmän kautta. Suuri osa lämmöstä poistuu sydänsulasta sen yläpuolella olevaan veteen, mutta lämmönsiirron tehostamiseksi myös sydänsiepparin alapuolelle on sijoitettu vedellä täytettävät jäähdytyskanavat. Jotta sydänsiepparin toiminta voitaisiin todentaa, on Lappeenrannan Teknillisellä Yliopistolla rakennettu Volley-koelaitteisto tätä tarkoitusta varten. Koelaitteisto koostuu kahdesta täysimittaisesta valuraudasta tehdystä jäähdytyskanavasta. Sydänsulan tuottamaa jälkilämpöä simuloidaan koelaitteistossa sähkövastuksilla. Tässä työssä kuvataan simulaatioiden suorittaminen ja vertaillaan saatuja arvoja mittaustuloksiin. Työ keskittyy sydänsiepparista jäähdytyskanaviin tapahtuvan lämmönsiirron teoriaan jamekanismeihin. Työssä esitetään kolme erilaista korrelaatiota lämmönsiirtokertoimille allaskiehumisen tapauksessa. Nämä korrelaatiot soveltuvat erityisesti tapauksiin, joissa vain muutamia mittausparametreja on tiedossa. Työn toinen osa onVolley 04 -kokeiden simulointi. Ensin käytettyä simulointitapaa on kelpoistettuvertaamalla tuloksia Volley 04 ja 05 -kokeisiin, joissa koetta voitiin jatkaa tasapainotilaan ja joissa jäähdytteen käyttäytyminen jäähdytyskanavassa on tallennettu myös videokameralla. Näiden simulaatioiden tulokset ovat hyvin samanlaisiakuin mittaustulokset. Korkeammilla lämmitystehoilla kokeissa esiintyi vesi-iskuja, jotka rikkoivat videoinnin mahdollistavia ikkunoita. Tämän johdosta osassa Volley 04 -kokeita ikkunat peitettiin metallilevyillä. Joitakin kokeita jouduttiin keskeyttämään laitteiston suurten lämpöjännitysten johdosta. Tällaisten testien simulaatiot eivät ole yksinkertaisia suorittaa. Veden pinnan korkeudesta ei ole visuaalista havaintoa. Myöskään jäähdytteen tasapainotilanlämpötiloista ei ole tarkkaa tietoa, mutta joitakin oletuksia voidaan tehdä samoilla parametreilla tehtyjen Volley 05 -kokeiden perusteella. Mittaustulokset Volley 04 ja 05 -kokeista, jotka on videoitu ja voitu ajaa tasapainotilaan saakka, antoivat simulaatioiden kanssa hyvin samankaltaisia lämpötilojen arvoja. Keskeytettyjen kokeiden ekstrapolointi tasapainotilaan ei onnistunut kovin hyvin. Kokeet jouduttiin keskeyttämään niin paljon ennen termohydraulista tasapainoa, ettei tasapainotilan reunaehtoja voitu ennustaa. Videonauhoituksen puuttuessa ei veden pinnan korkeudesta saatu lisätietoa. Tuloksista voidaan lähinnä esittää arvioita siitä, mitä suuruusluokkaa mittapisteiden lämpötilat tulevat olemaan. Nämä lämpötilat ovat kuitenkin selvästi alle sydänsiepparissa käytettävän valuraudan sulamislämpötilan. Joten simulaatioiden perusteella voidaan sanoa, etteivät jäähdytyskanavien rakenteet sula, mikäli niissä on pienikin jäähdytevirtaus, eikä useampia kuin muutama vierekkäinen kanava ole täysin kuivana.

Numerical and ExperimentalModelling of Gas Flow and Heat Transfer in the Air Gap of An Electric Machine

This work deals with the cooling of high-speed electric machines, such as motors and generators, through an air gap. It consists of numerical and experimental modelling of gas flow and heat transfer in an annular channel. Velocity and temperature profiles are modelled in the air gap of a high-speed testmachine. Local and mean heat transfer coefficients and total friction coefficients are attained for a smooth rotor-stator combination at a large velocity range. The aim is to solve the heat transfer numerically and experimentally. The FINFLO software, developed at Helsinki University of Technology, has been used in the flow solution, and the commercial IGG and Field view programs for the grid generation and post processing. The annular channel is discretized as a sector mesh. Calculation is performed with constant mass flow rate on six rotational speeds. The effect of turbulence is calculated using three turbulence models. The friction coefficient and velocity factor are attained via total friction power. The first part of experimental section consists of finding the proper sensors and calibrating them in a straight pipe. After preliminary tests, a RdF-sensor is glued on the walls of stator and rotor surfaces. Telemetry is needed to be able to measure the heat transfer coefficients at the rotor. The mean heat transfer coefficients are measured in a test machine on four cooling air mass flow rates at a wide Couette Reynolds number range. The calculated values concerning the friction and heat transfer coefficients are compared with measured and semi-empirical data. Heat is transferred from the hotter stator and rotor surfaces to the coolerair flow in the air gap, not from the rotor to the stator via the air gap, althought the stator temperature is lower than the rotor temperature. The calculatedfriction coefficients fits well with the semi-empirical equations and precedingmeasurements. On constant mass flow rate the rotor heat transfer coefficient attains a saturation point at a higher rotational speed, while the heat transfer coefficient of the stator grows uniformly. The magnitudes of the heat transfer coefficients are almost constant with different turbulence models. The calibrationof sensors in a straight pipe is only an advisory step in the selection process. Telemetry is tested in the pipe conditions and compared to the same measurements with a plain sensor. The magnitudes of the measured data and the data from the semi-empirical equation are higher for the heat transfer coefficients than thenumerical data considered on the velocity range. Friction and heat transfer coefficients are presented in a large velocity range in the report. The goals are reached acceptably using numerical and experimental research. The next challenge is to achieve results for grooved stator-rotor combinations. The work contains also results for an air gap with a grooved stator with 36 slots. The velocity field by the numerical method does not match in every respect the estimated flow mode. The absence of secondary Taylor vortices is evident when using time averagednumerical simulation.

Use of physiological constraints to identify quantitative design principles for gene expression in yeast adaptation to heat shock

Background: Understanding the relationship between gene expression changes, enzyme activity shifts, and the corresponding physiological adaptive response of organisms to environmental cues is crucial in explaining how cells cope with stress. For example, adaptation of yeast to heat shock involves a characteristic profile of changes to the expression levels of genes coding for enzymes of the glycolytic pathway and some of its branches. The experimental determination of changes in gene expression profiles provides a descriptive picture of the adaptive response to stress. However, it does not explain why a particular profile is selected for any given response. Results: We used mathematical models and analysis of in silico gene expression profiles (GEPs) to understand how changes in gene expression correlate to an efficient response of yeast cells to heat shock. An exhaustive set of GEPs, matched with the corresponding set of enzyme activities, was simulated and analyzed. The effectiveness of each profile in the response to heat shock was evaluated according to relevant physiological and functional criteria. The small subset of GEPs that lead to effective physiological responses after heat shock was identified as the result of the tuning of several evolutionary criteria. The experimentally observed transcriptional changes in response to heat shock belong to this set and can be explained by quantitative design principles at the physiological level that ultimately constrain changes in gene expression. Conclusion: Our theoretical approach suggests a method for understanding the combined effect of changes in the expression of multiple genes on the activity of metabolic pathways, and consequently on the adaptation of cellular metabolism to heat shock. This method identifies quantitative design principles that facilitate understating the response of the cell to stress.

Heat shock response in yeast involves changes in both transcription rates and mRNA stabilities

We have analyzed the heat stress response in the yeast Saccharomyces cerevisiae by determining mRNA levels and transcription rates for the whole transcriptome after a shift from 25uC to 37uC. Using an established mathematical algorithm, theoretical mRNA decay rates have also been calculated from the experimental data. We have verified the mathematical predictions for selected genes by determining their mRNA decay rates at different times during heat stress response using the regulatable tetO promoter. This study indicates that the yeast response to heat shock is not only due to changes in transcription rates, but also to changes in the mRNA stabilities. mRNA stability is affected in 62% of the yeast genes and it is particularly important in shaping the mRNA profile of the genes belonging to the environmental stress response. In most cases, changes in transcription rates and mRNA stabilities are homodirectional for both parameters, although some interesting cases of antagonist behavior are found. The statistical analysis of gene targets and sequence motifs within the clusters of genes with similar behaviors shows that both transcriptional and post-transcriptional regulons apparently contribute to the general heat stress response by means of transcriptional factors and RNA binding proteins.

Consensus recommendations on training and competing in the heat.

Exercising in the heat induces thermoregulatory and other physiological strain that can lead to impairments in endurance exercise capacity. The purpose of this consensus statement is to provide up-to-date recommendations to optimize performance during sporting activities undertaken in hot ambient conditions. The most important intervention one can adopt to reduce physiological strain and optimize performance is to heat acclimatize. Heat acclimatization should comprise repeated exercise-heat exposures over 1-2 weeks. In addition, athletes should initiate competition and training in a euhydrated state and minimize dehydration during exercise. Following the development of commercial cooling systems (e.g., cooling vest), athletes can implement cooling strategies to facilitate heat loss or increase heat storage capacity before training or competing in the heat. Moreover, event organizers should plan for large shaded areas, along with cooling and rehydration facilities, and schedule events in accordance with minimizing the health risks of athletes, especially in mass participation events and during the first hot days of the year. Following the recent examples of the 2008 Olympics and the 2014 FIFA World Cup, sport governing bodies should consider allowing additional (or longer) recovery periods between and during events for hydration and body cooling opportunities when competitions are held in the heat.

Sprint performance under heat stress: A review.

Training and competition in major track-and-field events, and for many team or racquet sports, often require the completion of maximal sprints in hot (>30 °C) ambient conditions. Enhanced short-term (<30 s) power output or single-sprint performance, resulting from transient heat exposure (muscle temperature rise), can be attributed to improved muscle contractility. Under heat stress, elevations in skin/core temperatures are associated with increased cardiovascular and metabolic loads in addition to decreasing voluntary muscle activation; there is also compelling evidence to suggest that large performance decrements occur when repeated-sprint exercise (consisting of brief recovery periods between sprints, usually <60 s) is performed in hot compared with cool conditions. Conversely, poorer intermittent-sprint performance (recovery periods long enough to allow near complete recovery, usually 60-300 s) in hotter conditions is solely observed when exercise induces marked hyperthermia (core temperature >39 °C). Here we also discuss strategies (heat acclimatization, precooling, hydration strategies) employed by "sprint" athletes to mitigate the negative influence of higher environmental temperatures.

Use of the heat dissipation method for sap flow measurement in citrus nursery trees1

Sap flow could be used as physiological parameter to assist irrigation of screen house citrus nursery trees by continuous water consumption estimation. Herein we report a first set of results indicating the potential use of the heat dissipation method for sap flow measurement in containerized citrus nursery trees. 'Valencia' sweet orange [Citrus sinensis (L.) Osbeck] budded on 'Rangpur' lime (Citrus limonia Osbeck) was evaluated for 30 days during summer. Heat dissipation probes and thermocouple sensors were constructed with low-cost and easily available materials in order to improve accessibility of the method. Sap flow showed high correlation to air temperature inside the screen house. However, errors due to natural thermal gradient and plant tissue injuries affected measurement precision. Transpiration estimated by sap flow measurement was four times higher than gravimetric measurement. Improved micro-probes, adequate method calibration, and non-toxic insulating materials should be further investigated.

Application of Information Statistical Theory to the Description of the Effect of Heat Conduction on the Chemical Reaction Rate in Gases

The effect of the heat flux on the rate of chemical reaction in dilute gases is shown to be important for reactions characterized by high activation energies and in the presence of very large temperature gradients. This effect, obtained from the second-order terms in the distribution function (similar to those obtained in the Burnett approximation to the solution of the Boltzmann equation), is derived on the basis of information theory. It is shown that the analytical results describing the effect are simpler if the kinetic definition for the nonequilibrium temperature is introduced than if the thermodynamic definition is introduced. The numerical results are nearly the same for both definitions