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PubMed1986

Possible cytoskeletal association of 69,000- and 68,000-dalton heat shock proteins and structural relations among heat shock proteins in murine mastocytoma cells.

When murine mastocytoma cells (FMA 1) were heat shocked (42 degrees C for 4 h), nine heat shock proteins (HSPs) were detected by two-dimensional gel electrophoresis. Their apparent molecular weights were 100, 85, 69, 68, 32, 30, and 23 kDa (3 of 23 kDa). The structural homology of 4, 69, 68, 32, and 30 kDa, was demonstrated by two-dimensional tryptic peptide mapping. The 69- and 68-kDa HSPs were purified and rabbit antisera against these HSPs were prepared. A small fraction (less than 10%) of the 69- and 68-kDa HSPs were copurified with the microtubules and were present in the Triton X-100/KCl cytoskeletal fraction as shown by immunoblotting with the antiserum and by peptide mapping. Our results are consistent with the hypothesis of a cytoskeletal role for HSPs.

Animals↗
PubMed1996

The heat shock response and major heat shock proteins of Tritrichomonas mobilensis and Tritrichomonas augusta.

The responses to heat shock in Tritrichomonas mobilensis, a squirrel monkey parasite and Tritrichomonas augusta, an amphibian trichomonad, were evaluated by means of metabolic labeling with [35S]methionine. Electrophoretically separated trichomonad proteins synthesized at different temperatures were visualized by autoradiography and the label incorporation quantitated by a trichloroacetic acid precipitation procedure. A considerable difference in thermotolerance between the two species was found as the protein synthesis reached a maximum at 41 C in T. mobilensis and 37 C in T. augusta. The latter tolerated temperature increases 13 C above normal cultivation temperatures as compared to only 4 C thermotolerance range above normal in T. mobilensis. Major heat shock proteins (Hsps) were expressed in both T. mobilensis (with apparent Mr 94, 72, and 58 kDa) and T. augusta (Mr 94, 70, and 56 kDa) as revealed by autoradiography. Western blot analysis with polyclonal antibody against DnaK of Escherichia coli showed the presence of antigenic Hsp70 homologs in both trichomonads. Similarly, a polyclonal antibody against Hsp60 with broad interspecies cross-reactivity detected Hsp60 homologs in both T. mobilensis and T. augusta. The anti-DnaK antibody cross-reacted with a T. mobilensis protein localized in Golgi apparatus as demonstrated by immunoelectron microscopy. Immunocytochemistry on trichomonad frozen sections revealed the presence of the Hsp60 homolog in light-microscopic granules corresponding to hydrogenosomes.

Animals↗
PubMed1996

Construction of cDNA library from liver RNA of heat shocked rats and DNA sequence analysis of the clone containing the 3'-end untranslated region (3'UTR) of the heat inducible gene hsp70.2.

cDNA library constructed from liver RNA of rats subjected to hyperthermia was used to isolate divergent 3' end untranslated regions (3'UTR) of heat inducible hsp70.1 and hsp70.2 genes. As a result of a double selection procedure with the use of DNA-DNA hybridization and PCR analysis 9 clones containing cDNA sequences derived from the 3'UTR of the hsp70.2 gene were selected. Nucleotide sequence of the cloned inserts was established and the Northern blot analysis was performed to identify the heat inducible transcript encoded by the hsp70.2 gene.

Animals↗
PubMed2002

Interaction between Arabidopsis heat shock transcription factor 1 and 70 kDa heat shock proteins.

The activity of the Arabidopsis heat shock transcription factor (HSF) is repressed at normal conditions but activated by cellular stresses. Circumstantial evidence suggests that HSP70 may function as a negative feedback regulator of HSF activity. Here the interaction between HSF and HSP70 is reported using electrophoretic mobility shift and yeast two-hybrid assays. Subdomain mapping indicates an interaction of the activation domain and DNA-binding domain of HSF1 with HSP70.

Arabidopsis↗
PubMed1999

Heat shock proteins and heat shock protein-antibody complexes in placental tissues.

OBJECTIVE: The relationship between pregnancy outcome and expression of the heat shock proteins (hsps) or hsp-antibody complexes of 60kD (hsp60), 70kD (hsp70), and 90kD (hsp90) in placental tissue and circulating antibodies to hsps was evaluated. METHOD: Expression of hsp60, hsp70, and hsp90 in placentae from 12 women with preterm birth, eight with intrauterine growth restriction (IUGR), and 10 with term birth, as well as the presence of the corresponding antibodies, was investigated by a new carbocyanine double fluorescence technique. Results were compared with microbiological findings and circulating antibodies to hsps in sera. RESULTS: In each placental specimen examined, hsp60, hsp70, and hsp90 were identified. However, hsp70-antibody complexes were detected in only four of the preterm labor cases. Similarly, hsp60-antibody complexes were detected in only five preterm labor patients and in one patient with IUGR. None of the placentae contained hsp90-antibody complexes. In the preterm birth group, all patients with hsp60-antibody complexes were also positive for circulating antibodies to hsp60. The presence of hsp70-antibody complexes also correlated with hsp70 antibody in sera. CONCLUSIONS: Formation of hsp60- and hsp70-antibody complexes in the placenta may contribute to the induction of preterm birth. Women sensitized to these antibodies may be at increased risk for adverse pregnancy outcome.

Antigen-Antibody Complex↗
PubMed2006

Involvement of calcium in the differential induction of heat shock protein 70 by heat shock protein 90 inhibitors, geldanamycin and radicicol, in human non-small cell lung cancer H460 cells.

Both geldanamycin (GA) and radicicol (RA) are HSP90 binding agents that possess antitumour activities. Although the in vitro data indicated that the inhibitory constant of RA is much bigger than that of GA, the in vivo data on drug efficacy might reveal different results. We have recently shown that treatment with GA induces a heat-shock response and that calcium mobilization may be involved in the process. By using induction of HSP70 as the endpoint assay, we found changes in upstream signaling mediators, including HSF1 and calcium mobilization, as well as possible involvement of protein kinase in human non-small cell lung cancer H460 cells treated with GA and RA. Our results demonstrated that calcium mobilization, a calcium dependent and H7-sensitive protein kinase, along with HSF1 activation by phosphorylation, are all involved in the HSP70 induction process triggered by the drugs. However, only GA, but not RA, can provoke a rapid calcium mobilization and thereby result in an instant induction of HSP70. Furthermore, the rapid calcium influx, followed by instant HSP induction, could be achieved in GA- or RA-treated cells placed in a medium containing excessive calcium while the response was completely abolished in cells depleted of calcium. Taken together, our findings suggest that differential calcium signaling may account for the differential induction of HSP and the action of GA and RA.

Benzoquinones↗
PubMed2004

Heat stress response in plants: a complex game with chaperones and more than twenty heat stress transcription factors.

Compared to the overall multiplicity of more than 20 plant Hsfs, detailed analyses are mainly restricted to tomato and Arabidopsis and to three important representatives of the family (Hsfs A1, A2 and B1). The three Hsfs represent examples of striking functional diversification specialized for the three phases of the heat stress (hs) response (triggering, maintenance and recovery). This is best illustrated for the tomato Hsf system: (i) HsfA1a is the master regulator responsible for hs-induced gene expression including synthesis of HsfA2 and HsfB1. It is indispensible for the development of thermotolerance. (ii) Although functionally equivalent to HsfA1a, HsfA2 is exclusively found after hs induction and represents the dominant Hsf, the "working horse" of the hs response in plants subjected to repeated cycles of hs and recovery in a hot summer period. Tomato HsfA2 is tightly integrated into a network of interacting proteins (HsfA1a, Hsp17-CII, Hsp17-CI) influencing its activity and intracellular distribution. (iii) Because of structural peculiarities, HsfB1 acts as coregulator enhancing the activity of HsfA1a and/or HsfA2. But in addition, it cooperates with yet to be identified other transcription factors in maintaining and/or restoring housekeeping gene expression.

Amino Acid Sequence↗
PubMed1999

Polymorphism in the regulatory sequence of the human hsp70-1 gene does not affect heat shock factor binding or heat shock protein synthesis.

A bi-allelic polymorphism found in the regulatory region of the human heat shock (HS) protein (HSP) hsp70-1 gene, which comprises an A-->C transversion, 3 bp upstream of the HS element (HSE), has been associated with extended HLA haplotypes. In view of the chaperoning and protective functions of Hsp70, we investigated whether this hsp70-1 bi-allelic polymorphism could modulate the stress response, which may relate to enhanced resistance or susceptibility to certain diseases. We compared the basal and HS-induced HS factor (HSF)-binding activity of the two polymorphic HSEs, hsp70-1 mRNA accumulation and HSP expression in two human Epstein Barr virus (EBV)-transformed B cell lines typed for hsp70-1 promoter alleles. Our results suggest that hsp70-1 promoter polymorphism does not influence HSF-binding activity, hsp70 mRNA accumulation or synthesis in human EBV-transformed B cell lines.

Alleles↗
PubMed2001

Mediator, not holoenzyme, is directly recruited to the heat shock promoter by HSF upon heat shock.

Activators of RNA polymerase II (Pol II) transcription have been shown to bind several coactivators and basal factors in vitro. Whether such interactions play a primary regulatory role in recruiting these factors to activator-associated chromosomal target sites in living cells remains unclear. Here, we show that upon heat shock the Pol II-free form of Mediator is rapidly recruited to HSF binding sites. Unlike the TAFs and Pol II, the interaction between Mediator and HSF on chromosomal loci is direct and mechanistically separable from the preinitiation complex assembly step. Therefore, the activator-Mediator interaction likely underlies the initiation of signal transfer from enhancer-bound activators to the basal transcription machinery.

Amanitins↗
PubMed2001

Arabidopsis and the heat stress transcription factor world: how many heat stress transcription factors do we need?

Sequencing of the Arabidopsis genome revealed a unique complexity of the plant heat stress transcription factor (Hsf) family. By structural characteristics and phylogenetic comparison, the 21 representatives are assigned to 3 classes and 14 groups. Particularly striking is the finding of a new class of Hsfs (AtHsfC1) closely related to Hsf1 from rice and to Hsfs identified from frequently found expressed sequence tags of tomato, potato, barley, and soybean. Evidently, this new type of Hsf is well expressed in different plant tissues. Besides the DNA binding and oligomerization domains (HR-A/B region), we identified other functional modules of Arabidopsis Hsfs by sequence comparison with the well-characterized tomato Hsfs. These are putative motifs for nuclear import and export and transcriptional activation (AHA motifs). There is intriguing flexibility of size and sequence in certain parts of the otherwise strongly conserved N-terminal half of these Hsfs. We have speculated about possible exon-intron borders in this region in the ancient precursor gene of plant Hsfs, similar to the exon-intron structure of the present mammalian Hsf-encoding genes.

Amino Acid Sequence↗
PubMed1991

Heated fats. Part 3. Biological effect and effect of heating and tempering oils on fatty acid composition of liver, heart and serum lipids of rats.

The study deals with the biological effect caused by changes in fats during heating. The study includes feeding experiments and extraction of serum, liver, and heart from the animals tested. The biological study reveals that animals fed heated oil showed retardation of growth, poor efficiencies, rough, greasy mottled coats, and shortened life span.

Animals↗
PubMed1992

Stimulation of the nigrostriatal dopamine system inhibits both heat production and heat loss mechanisms in rats.

The effects of stimulating the pars compacta of the substantia nigra (SNC) on thermoregulation were assessed in normal rats, in rats with chemical lesion of the SNC dopamine (DA) pathways and in rats with striatal DA receptor blockade. Electrical stimulation of the SNC produced hypothermia, decreased metabolism and/or cutaneous vasoconstriction in rats at ambient temperatures (Ta) below 22 degrees C, as well as hyperthermia and cutaneous vasoconstriction in rats at Ta of 30 degrees C. Microinjection of an excitotoxic amino acid (kainic acid) at the same brain sites also produced the same thermal responses. In vivo voltammetric studies revealed that electrical or chemical stimulation of the SNC produced an increase in striatal DA release. The enhanced striatal DA release induced by SNC stimulation was attenuated in rats after selective destruction of the nigrostriatal DA pathway by administration of 6-hydroxydopamine into the medial forebrain bundle. In addition, the magnitude of the thermal responses produced by the SNC stimulation in the cold was attenuated by selective bilateral destruction of the nigrostriatal DA pathways or selective blockade of the striatal DA produced by intrastriatal infusion of haloperidol, a DA receptor antagonist. The results indicate that stimulation of the SNC inhibits both heat production and heat loss mechanisms in the rat.

Animals↗
PubMed1988

A Neurospora crassa heat-shocked cell lysate translates homologous and heterologous messenger RNA efficiently, without preference for heat shock messages.

Cell-free protein synthesis systems were prepared from normally-grown (N-lysate) and heat-shocked (HS-lysate) Neurospora crassa mycelium. Although both lysates translated homologous mRNA, the HS-lysate was more active, yielding a higher incorporation of [35S]-methionine into hot TCA-insoluble material and a vastly superior protein synthesis profile. The optimal temperature for translation by both lysates was 21 degrees C; the HS-lysate did not translate heat-shock mRNA preferentially at any temperature tested. Fortuitously, heterologous messenger RNAs from diverse eukaryotic and viral sources - Drosophila, dog pancreas, rabbit globin mRNA, brome mosaic virus, tobacco mosaic virus - were translated by the HS-lysate with an efficiency comparable to that of the commercial rabbit reticulocyte system and superior to the wheat germ system. The cap analogues, m7G(5')ppp(5')G and m7G(5')Gm, inhibited translation significantly.

Animals↗
PubMed1990

Solar heat load: heat balance during exercise in clothed subjects.

Six subjects exercised for 60 min on a cycle ergometer. Their backs were exposed to an artificial 'sun' with a spectral distribution similar to sunlight and an intensity of 724 W m-2. Each subject took part in four experiments in random order: wearing suits of polyester (insulation value = 0.5 clo), white (WP) or black (BP), or cotton (0.6 clo), white (WC) or black (BC). Measured by partitional calorimetry, the calculated heat losses and gains for the four conditions balanced within less than 10%. The differences between the short-wave radiation gains of subjects in white or black garments were small. This is due to the transparency of the white materials, which allows a larger percentage of the radiation to penetrate the clothing. The surface temperatures of the sun-exposed areas were very high, especially in the black suits. This promotes dry heat loss. Therefore the sweat loss in the black suits and the differences between the black and white clothes became relatively small. The physiological strain in steady-state exercise, as expressed by average heart rates, was 142 (WP), 154 (BP), 151 (WC), and 160 (BC) beats min-1; the sweat losses were 649 (WP), 666 (BP), 704 (WC), and 808 (BC) g. For both of these measures values for white polyester were significantly less than those for black cotton.

Adult↗
PubMed1978

Model studies on the heating of food proteins--heat-induced oligomerisation of ribonuclease.

Commercial ribonuclease was heated at temperatures between 80 degrees C and 180 degrees C for 1--24 h. A stoichiometric oligomerisation was observed. Dimeric ribonuclease appeared after 1 h at 80 degrees C. With increasing time and/or temperature the number of oligomers formed rose at first. The highest oligomer that could be detected was the hexamer (140 degrees C, 8 and 16 h). A further increase in time and/or temperature resulted in a decrease of the number of oligomers and a rise in the polymer fraction. The importance of this reaction in the changes produced by heating food proteins and various reaction mechanisms are discussed.

Dietary Proteins↗
PubMed1992

Comparison of hydrophobic heat and moisture exchangers with heated humidifier during prolonged mechanical ventilation.

Inspired gases must be warmed and humidified during mechanical ventilation. In a prospective randomized study we compared the performance of a heated humidifier (HH) (Draegger Aquaport) and a heat and moisture exchanger (HME) (Pall Filter BB 2215). A total of 116 patients requiring mechanical ventilation (Servo 900 C Siemens) were enrolled into the study and were randomly assigned to 2 groups. Patients in group I were ventilated with a traditional breathing circuit with HH and patients in group II using a simplified circuit with HME. Pre-existing and hospital acquired atelectasis and pneumonia, occurrence of endotracheal tube (ET) occlusion and ventilatory parameters (respiratory rate, tidal volume) were studied. No statistical difference was found between groups for each parameter except the greater frequency of ET occlusions in the II group (0/61 vs 9/55) (p = 0.0008). Pall Filter (PF), a hydrophobic filter, humidifies the dry gases from the condensed water which is put down on the HME surfaces during cooling of saturated expired gases. This purely physical property is linked to the magnitude of the thermic gradient between the expired gases and the ambiant temperature. Performance impairment of PF in our study might be due to high ambiant temperature in the intensive care unit (usually around 28 degrees C) which reduces thermic gradient and water exchanges. We conclude that efficiency of PF may be weak in some conditions of ambiant temperature.

Adult↗
PubMed1975

Dry-heat inactivation of Bacillus subtilis spores by means of infra-red heating.

An experumental equipment for dry-heat inactivation of bacterial spores in an open system using Infrared (IR) radiation for energy transfer was deveoped. The dry-heat-inactivation kinetics for Bacillus subtilis ATCC 6633 spores were studied in the temperature range of 120-180 C. The z value (z = 23C) was constant in the temperature range investigated. The advantages offered by using IR radiation in sterilization systems are pointed out.

Bacillus subtilis↗
PubMed2002

Comparison of the effects of heat and moisture exchangers and heated humidifiers on ventilation and gas exchange during non-invasive ventilation.

OBJECTIVE: To compare the short-term effects of a heat and moisture exchanger (HME) and a heated humidifier (HH) during non-invasive ventilation (NIV). DESIGN: Prospective, clinical investigation. SETTING: Intensive care unit of a university hospital. PATIENTS: Twenty-four patients with acute respiratory failure (ARF). INTERVENTION: Each patient was studied with a HME and a HH in a random order during two consecutive 20min periods of NIV. MEASUREMENTS AND RESULTS: Respiratory rate (RR), expiratory tidal volume (VTe) and expiratory minute ventilation (VE) were measured during the last 5 min of each period and blood gases were measured. Mean pressure support and positive end-expiratory pressure levels were, respectively, 15+/-4 and 6+/-2 cmH(2)O. VE was significantly greater with HME than with HH (14.8+/-4.8 vs 13.2+/-4.3 l/min; p<0.001). This increase in VE was the result of a greater RR for HME than for HH (26.5+/-10.6 vs 24.1+/-9.8 breaths/min; p=0.002), whereas the VT for HME was similar to that for HH (674+/-156 vs 643+/-148 ml; p=0.09). Arterial partial pressure of carbon dioxide (PaCO(2)) was significantly higher with a HME than with a HH (43.4+/-8.9 vs 40.8+/-8.2 mmHg; p<0.005), without significantly changing oxygenation. CONCLUSION: During NIV the increased dead space of a HME can negatively affect ventilatory function and gas exchange. The effect of HME dead space may decrease efficiency of NIV in patients with ARF.

Acute Disease↗