Shock Protection Research Articles

Chlamydia trachomatis responds to heat shock, penicillin induced persistence, and IFN-gamma persistence by altering levels of the extracytoplasmic stress response protease HtrA

BackgroundChlamydia trachomatis, an obligate intracellular human pathogen, is the most prevalent bacterial sexually transmitted infection worldwide and a leading cause of preventable blindness. HtrA is a virulence and stress response periplasmic serine protease and molecular chaperone found in many bacteria. Recombinant purified C. trachomatis HtrA has been previously shown to have both activities. This investigation examined the physiological role of Chlamydia trachomatis HtrA.ResultsThe Chlamydia trachomatis htrA gene complemented the lethal high temperature phenotype of Escherichia coli htrA- (>42°C). HtrA levels were detected to increase by western blot and immunofluorescence during Chlamydia heat shock experiments. Confocal laser scanning microscopy revealed a likely periplasmic localisation of HtrA. During penicillin induced persistence of Chlamydia trachomatis, HtrA levels (as a ratio of LPS) were initially less than control acute cultures (20 h post infection) but increased to more than acute cultures at 44 h post infection. This was unlike IFN-γ persistence where lower levels of HtrA were observed, suggesting Chlamydia trachomatis IFN-γ persistence does not involve a broad stress response.ConclusionThe heterologous heat shock protection for Escherichia coli, and increased HtrA during cell wall disruption via penicillin and heat shock, indicates an important role for HtrA during high protein stress conditions for Chlamydia trachomatis.

Heat Shock Proteins and Protection of the Nervous System

Manipulation of the cellular stress response offers strategies to protect brain cells from damage induced by ischemia and neurodegenerative diseases. Overexpression of Hsp70 reduced ischemic injury in the mammalian brain. Investigation of the domains within Hsp70 that confers ischemic neuroprotection revealed the importance of the carboxyl-terminal domain. Arimoclomol, a coinducer of heat shock proteins, delayed progression of amyotrophic lateral sclerosis (ALS) in a mouse model in which motor neurons in the spinal cord and motor cortex degenerate. Celastrol, a promising candidate as an agent to counter neurodegenerative diseases, induced expression of a set of Hsps in differentiated neurons grown in tissue culture. Heat shock "preconditioning" protected the nervous system at the functional level of the synapse and selective overexpression of Hsp70 enhanced the level of synaptic protection. Following hyperthermia, constitutively expressed Hsc70 increased in synapse-rich areas of the brain where it associates with Hsp40 to form a complex that can refold denatured proteins. Stress tolerance in neurons is not solely dependent on their own Hsps but can be supplemented by Hsps from adjacent glial cells. Hence, application of exogenous Hsps at neural injury sites is an effective strategy to maintain neuronal viability.

Geldanamycin, an inhibitor of Hsp90, Blocks cytoplasmic retention of progesterone receptors and glucocorticoid receptors via their respective ligand binding domains

Steroid hormone receptors (SHRs), such as glucocorticoid receptors (GR) and progesterone receptors (PR), are shuttling proteins that undergo continuous nuclear import and export. Various mechanisms have been proposed to explain the localization of SHRs. It has been suggested that the ligand-binding domain (LBD) of SHRs is important in determining the subcellular localization. We have studied the localization of GR-LBD and PR-LBD alone, as well as of full-length GR and PR in the presence of geldanamycin (GA), a benzoquinoid ansamycin that specifically inhibits heat shock protection (Hsp90), using transient transfections and fluorescent microscopy. Our studies have indicated that GR-LBD and PR-LBD are retained in the cytoplasm via interaction with Hsp90. It was observed that in the unliganded state, treatment with GA translocates these LBDs to the nucleus. Similar results were obtained for full-length PR and GR. Additionally, it was found that after ligand induction, GA accelerated reexport of SHRs after ligand washout, implicating Hsp90 in nuclear retention of SHRs in the washout state. We also propose that a recently found "export" signal present in the LBD of SHRs is involved in interactions with Hsp90 and hence cytoplasmic retention of these receptors. After ligand induction, Hsp90 also may play a role in nuclear retention of SHRs following hormone washout.

Design and Analysis of a Disk Drive Actuator for Commutational Ramp Loading

A voice-coil motor actuator system is designed to perform a unique disk drive ramp load/unload operation. The design eliminates the necessity of increased material requirements common in ramp load disk drives. Therefore, disk drives with lower cost, higher performance actuators can realize the linear shock protection benefits of ramp loading. This study offers a complete, systematic design procedure together with required analysis for manufacture of a commutational ramp load/unload drive. The actuator is designed and optimized to meet specific move-time performance requirements. When used for ramp load/unload, however, there exists a location where input current has no influence on actuator motion. An input polarity reversal is required within the uncontrollable region to sustain the direction of actuator motion. A magnetic, restorative bias is designed that prevents the actuator from resting in the uncontrollable region while providing resistance to rotational shock effects. A state trajectory is designed that, when tracked, moves the actuator through the uncontrollable set for a successful load onto the disk at the desired load velocity. A ramp load controller is designed to track the trajectory and handle the nonlinear effects from bias and friction torque. The unique disk drive is manufactured and experiments are performed to demonstrate effectiveness of the complete design strategy.

Micromachined integrated shock protection for MEMS

This paper reports the design and analysis of two concepts for shock protection of MEMS using nonlinear springs and soft coatings. The two concepts (nonlinear springs and soft coatings) could improve shock protection by reducing the secondary impact of a moveable MEMS device to shocks stops and by dissipating its energy in a reliable manner. We analyze the merits and demerits of each concept using simulation and ranges of shock design parameters are suggested to enhance performance. Simulations reveal that the nonlinear spring shock stops are capable of reducing impulses by over 90% while soft coatings may reduce impulses by 40%.

Induction of the 72-kilodalton heat shock protein and protection from ultraviolet B–induced cell death in human keratinocytes by repetitive exposure to heat shock or 15-deoxy-Δ12,14-prostaglandin J2

It has been demonstrated that hyperthermia protects keratinocytes from ultraviolet B (UVB)-induced cell death in culture and in vivo. This effect is mediated by the antiapoptotic effect of heat shock proteins that are transiently induced after exposure to heat at sublethal temperatures. Consequently, induction of Hsp has been proposed as a novel means of photoprotection. However, in the face of daily UVB exposure of human skin in vivo, this approach would not be useful if keratinocytes become less sensitive to Hsp induction with repeated exposure to the inducing agent. The aim of this study was to investigate whether repeated exposure to hyperthermia or to the stress protein activating cyclopentenone prostaglandin 15-deoxy-delta(12,14)-prostaglandin J2 (15dPGJ2) leads to adaptation of the cells, attenuation of the heat shock response, and abrogation of the protective effect. Normal human epidermal keratinocytes (NHEK) and the carcinoma-derived cell line A431 were exposed to either 42 degrees C or to 15dPGJ2 for 4 hours at 24-hour intervals for 4 consecutive days. The intracellular level of the 72-kDa heat shock protein (Hsp72) was determined by enzyme-linked immunosorbent assay (ELISA). Cells were exposed to UVB from a metal halide source after the last heat or 15dPGJ2 treatment, and survival was determined 24 hours after exposure by a MTT assay. Our results demonstrate that (1) heat shock and 15dPGJ2 are potent inducers of Hsp72 expression and lead to increased resistance to UVB-induced cell death in human keratinocytes; (2) re-exposure to heat shock leads to a superinduction without attenuation of the absolute increase in Hsp72 and of its UVB-protective effect; (3) the UVB tolerance induced by 15dPGJ2 is enhanced by repeated exposure without a further increase of Hsp72; (4) repeated heat shock and 15dPGJ2 up to a concentration of 1 microg/mL have no influence on cell growth over a period of 4 days. We conclude that through repeated exposure to Hsp-inducing factors, stress tolerance can be maintained without additional toxicity in human keratinocytes. These results provide a basis for the development of nontoxic Hsp inducers that can be repeatedly applied without loss of effect.

Global Transcriptional and Proteomic Analysis of the Sig1 Heat Shock Regulon ofDeinococcus radiodurans

The sig1 gene, predicted to encode an extracytoplasmic function-type heat shock sigma factor of Deinococcus radiodurans, has been shown to play a central role in the positive regulation of the heat shock operons groESL and dnaKJ. To determine if Sig1 is required for the regulation of additional heat shock genes, we monitored the global transcriptional and proteomic profiles of a D. radiodurans R1 sig1 mutant and wild-type cells in response to elevated temperature stress. Thirty-one gene products were identified that showed heat shock induction in the wild type but not in the sig1 mutant. Quantitative real-time PCR experiments verified the transcriptional requirement of Sig1 for the heat shock induction of the mRNA of five of these genes-dnaK, groES, DR1314, pspA, and hsp20. hsp20 appears to encode a new member of the small heat shock protein superfamily, DR1314 is predicted to encode a hypothetical protein with no recognizable orthologs, and pspA is predicted to encode a protein involved in maintenance of membrane integrity. Deletion mutation analysis demonstrated the importance in heat shock protection of hsp20 and DR1314. The promoters of dnaKJE, groESL, DR1314, pspA, and hsp20 were mapped and, combined with computer-based pattern searches of the upstream regions of the 26 other Sig1 regulon members, these results suggested that Sig1 might recognize both sigma70-type and sigma(W)-type promoter consensus sequences. These results expand the D. radiodurans Sig1 heat shock regulon to include 31 potential new members, including not only factors with cytoplasmic functions, such as groES and dnaK, but also those with extracytoplasmic functions, like pspA.

보안등의 감전 위험성에 관한 실태조사

본 논문에서는 보안등의 감전 위험성의 실태를 파악하기 위하여 세 지역에서 보안등을 조사하였다. 취득된 데이터에 의거하여 보안등의 실태를 감전 위험성 측면에서 정밀하게 분석하였다. 조사결과, 많은 보안등은 규정이 미비하여 올바르게 설치되어 있지 않기 때문에 감전의 위험성이 증가되는 것으로 밝혀졌다. 따라서 감전 예방의 관점에서 보안등 시설의 설치 및 관리에 대한 구체적인 규정을 설정하는 것이 필요하다.

FEA for designing of floating raft shock-resistant system

Choosing the equipment with good shock-resistant performance and taking shock protection measures while designing the onboard settings, the safety of onboard settings can be assured when warships, especially submarine subjected to non-contact underwater explosion, that is, these means can be used to limit the rattlespace (i. e., the maximum displacement of the equipment relative to the base) and the peak acceleration experienced by the equipment. Using shock-resistant equipments is one of shock protection means. The shock-resistant performance of the shock-resistant equipments should be verified in the design phase of the equipments. The FEA (finite element analysis) software, for example, MSC. NASTRAN®, can be used to verify the shock-resistant performance. MSC. PATRAN® and MSC. NASTRAN are used for modeling and analyzing the floating raft vibration isolating equipment. The model of the floating raft and the floating raft vibration isolating system are theoretically analyzed and calculated, and the analysis results are in agreement with the test results. The transient response analysis of the system model follows the modal analysis of the floating raft vibration isolating system. And it is used to verify the shock-resistant performance. The analysis and calculation method used in this paper can be used to analyze the shock-resistant performance of onboard shock-resistant equipments.

Heat Shock Suppresses the Permeability Transition in Rat Liver Mitochondria

Heat shock proteins inhibit apoptotic and necrotic cell death in various cell types. However, the specific mechanism underlying protection by heat shock proteins remains unclear. To test the hypothesis that heat shock proteins inhibit cell death by blocking opening of mitochondrial permeability transition (MPT) pores, mitochondria from heat-preconditioned rat livers were isolated by differential centrifugation. Heat shock inhibited MPT pore opening induced by 50 microm CaCl(2) plus 5 microm HgCl(2) or 1 microm mastoparan and by 200 microm CaCl(2) alone. Half-maximal swelling was delayed 15 min or more after heat shock compared with control. Heat shock also increased the threshold of unregulated (Ca(2+)-independent and cyclosporin A-insensitive) MPT pore opening induced by higher doses of HgCl(2) and mastoparan. Heat shock treatment decreased mitochondrial reactive oxygen species formation by 27% but did not change mitochondrial respiration, membrane potential, Ca(2+) uptake, or total glutathione in mitochondrial and cytosolic extracts of liver. Western blot analysis showed that mitochondrial Hsp25 increased, whereas Hsp10, Hsp60, Hsp70, Hsp75, cyclophilin D, and voltage-dependent anion channel did not change after heat shock. These results indicate that heat shock causes resistance to opening of MPT pores, which may contribute to heat shock protection against cellular injury.

Role of interleukin-6 in hepatic heat shock protein expression and protection against acetaminophen-induced liver disease

Recent experimental data suggest that the idiosyncratic nature of drug-induced liver disease (DILD) may be due in part to a deficiency of one or more hepatoprotective factors. In this study we have investigated whether interleukin (IL)-6 may also be one of these factors. Following the induction of liver injury with acetaminophen (APAP), a time-dependent increase in liver mRNA expression of IL-6 and its family members IL-11, leukemia inhibitory factor, and oncostatin M was observed in wild type (WT) mice, suggesting a possible hepatoprotective role played by this cytokine family. Indeed, mice lacking IL-6 ( IL-6 −/−) were more susceptible than were WT mice to APAP-induced liver injury. The increased susceptibility of the IL-6 −/− mice was associated with a deficiency in the expression of hepatic heat shock protein (HSP)25, 32, and 40 as well as inducible HSP70 following APAP treatment. These results suggest that IL-6 and possibly other family members may protect the liver from injury, at least in part, by up-regulating the hepatic expression of several cytoprotective HSPs.

Expression of a 72-kDa heat shock protein and protection in gastric mucosa of portal hypertensive rats

Expression of a 72-kDa heat shock protein and protection in gastric mucosa of portal hypertensive rats

Involvement of two putative alternative sigma factors in stress response of the radioresistant bacterium Deinococcus radiodurans.

Two genes bearing similarity to alternative sigma factors were identified in the Deinococcus radiodurans genome sequence and designated sig1 and sig2. These genes were cloned and inactivated, and both were found to be important for survival during heat and ethanol stress, although the sig1 mutants displayed a more severe phenotype than the sig2 mutants. Reporter gene fusions to the groESL and dnaKJ operons transformed into these mutant backgrounds indicated that sig1 is required for the heat shock induction of groESL and dnaKJ, whereas sig2 mutants show a more moderate defect in dnaKJ induction and are not impaired for groESL induction. Essentiality tests suggested that neither sig1 nor sig2 is essential under all conditions. Sequence comparisons demonstrated that the sig1 gene product is classed distinctly with extracytoplasmic function (ECF) sigma factors, whereas Sig2 appears to be a more divergent sigma factor ortholog. These results suggest that sig1 encodes the major ECF-derived heat shock sigma factor in D. radiodurans and that it plays a central role in the positive regulation of heat shock genes. sig2, in contrast, appears to play a more minor role in heat shock protection and may serve to modulate the expression of some heat protective genes.

Assessment of the noise exposure of call centre operators.

Call centres now play a major role in the daily operations of financial, technology and utility companies, as well as public bodies. It is predicted that 2002 will see 2.3% of the total British workforce employed in call centres. However, local authority enforcement officers, unions, voluntary organizations, employers and employees have all expressed concern that there are hazards to health and safety unique to this new and developing industry. One of the potential hazards reported in the press is hearing damage from using headsets. In a Health & Safety Executive funded project, the noise exposure of 150 call centre operators was evaluated, in call centres which included financial services, home shopping and telecommunications services. The results show that the daily personal noise exposure of these call centre operators is unlikely to exceed the 85 dB(A) action level defined in the Noise at Work Regulations 1989. The risk of hearing damage is therefore extremely low. Exposure to higher noise levels is possible, for example from fax tones, holding tones and high pitched tones from mobile phones. However, the duration of these events is likely to be short and they are therefore unlikely to have a significant effect on the operators' overall noise exposure. A practical method of limiting exposure to unexpected high noises from headsets is to ensure that the headsets incorporate acoustic shock protection that meets the requirements of the Department of Trade and Industry specification 85/013. In the UK, this limiter ensures any noise above 118 dB is not transmitted through the headset. Operators should receive regular training on the headset and telephone equipment they are using. This training should include correct use of the headset and the volume control facilities, and advice on how and when to clean and maintain the headsets.

Prolonged nuclear retention of activated extracellular signal-regulated protein kinase promotes cell death generated by oxidative toxicity or proteasome inhibition in a neuronal cell line.

In the HT22 mouse hippocampal cell line and primary immature embryonic rat cortical neurons, glutamate-induced oxidative toxicity is associated with a delayed but chronic activation of extracellular signal-regulated kinase-1/2 (ERK-1/2). ERK-1/2 is also activated in HT22 cells that undergo caspase-dependent cell death upon inhibition of proteasome-dependent protein degradation brought about by MG132 treatment. As in glutamate-treated HT22 cells and primary neurons, inhibition of MEK-1, an upstream activator of ERK-1/2 protects against MG132-induced toxicity. Furthermore, activated ERK-1/2 is retained within the nucleus in glutamate- and MG132-treated HT22 cells. Although previous studies suggested that ERK-1/2 activation was downstream of many cell death-inducing signals in HT22 cells, we show here that cycloheximide, the Z-vad caspase inhibitor, and a nonlethal heat shock protect against glutamate- and MG132-induced toxicity without diminishing ERK-1/2 activation. In these cases, ERK-1/2, although chronically activated, is not retained within the nucleus but accumulates within the cytoplasm. Thus, persistent nuclear retention of activated ERK-1/2 may be a critical factor in eliciting proapoptotic effects in neuronal cells subjected to oxidative stress or proteasome inhibition.

Accelerated metabolism and exclusion of 4-hydroxynonenal through induction of RLIP76 and hGST5.8 is an early adaptive response of cells to heat and oxidative stress.

To explore the role of lipid peroxidation (LPO) products in the initial phase of stress mediated signaling, we studied the effect of mild, transient oxidative or heat stress on parameters that regulate the cellular concentration of 4-hydroxynonenal (4-HNE). When K562 cells were exposed to mild heat shock (42 degrees C, 30 min) or oxidative stress (50 microM H2O2, 20 min) and allowed to recover for 2 h, there was a severalfold induction of hGST5.8, which catalyzes the formation of glutathione-4-HNE conjugate (GS-HNE), and RLIP76, which mediates the transport of GS-HNE from cells (Awasthi, S., Cheng, J., Singhal, S. S., Saini, M. K., Pandya, U., Pikula, S., Bandorowicz-Pikula, J., Singh, S. V., Zimniak, P., and Awasthi, Y. C. (2000) Biochemistry 39, 9327-9334). Enhanced LPO was observed in stressed cells, but the major antioxidant enzymes and HSP70 remained unaffected. The stressed cells showed higher GS-HNE-conjugating activity and increased efflux of GS-HNE. Stress-pre-conditioned cells with induced hGST5.8 and RLIP76 acquired resistance to 4-HNE and H2O2-mediated apoptosis by suppressing a sustained activation of c-Jun N-terminal kinase and caspase 3. The protective effect of stress pre-conditioning against apoptosis was abrogated by coating the cells with anti-RLIP76 IgG, which inhibited the efflux of GS-HNE from cells, indicating that the cells acquired resistance to apoptosis by metabolizing and excluding 4-HNE at a higher rate. Induction of hGST5.8 and RLIP76 by mild, transient stress and the resulting resistance of stress-pre-conditioned cells to apoptosis appears to be a general phenomenon since it was not limited to K562 cells but was also evident in lung cancer cells, H-69, H-226, human leukemia cells, HL-60, and human retinal pigmented epithelial cells. These results strongly suggest a role of LPO products, particularly 4-HNE, in the initial phase of stress mediated signaling.

The loop domain of heat shock transcription factor 1 dictates DNA-binding specificity and responses to heat stress.

Eukaryotic heat shock transcription factors (HSF) regulate an evolutionarily conserved stress-response pathway essential for survival against a variety of environmental and developmental stresses. Although the highly similar HSF family members have distinct roles in responding to stress and activating target gene expression, the mechanisms that govern these roles are unknown. Here we identify a loop within the HSF1 DNA-binding domain that dictates HSF isoform specific DNA binding in vitro and preferential target gene activation by HSF family members in both a yeast transcription assay and in mammalian cells. These characteristics of the HSF1 loop region are transposable to HSF2 and sufficient to confer DNA-binding specificity, heat shock inducible HSP gene expression and protection from heat-induced apoptosis in vivo. In addition, the loop suppresses formation of the HSF1 trimer under basal conditions and is required for heat-inducible trimerization in a purified system in vitro, suggesting that this domain is a critical part of the HSF1 heat-stress-sensing mechanism. We propose that this domain defines a signature for HSF1 that constitutes an important determinant for how cells utilize a family of transcription factors to respond to distinct stresses.

Heat shock proteins and protection against ischemic injury

Heat shock proteins present a complex family of proteins exerting chaperone-like activities that are classified according to their molecular weight. We especially explored protective functions of inducible heat shock protein 70, the mitochondrial heat shock protein 60 and 10, and the small heat shock proteins HSP27 and αB-crystallin against ischemic, reoxygenation-mediated injury using transgenic animals and hearts under in vivo conditions and in isolated cardiac myocyte-derived cells using adenoviral vectors. We noted with great interest that differential protective effects are exerted by specific hsps. For example, alpha-B-crystallin and constitutive hsp70 markedly protect microtubular structure in cardiac myocytes from ischemia-induced injury. Inducible hsp70, hsp60 and hspl0 when coexpressed, and hsp27 and αB-crystallin have an overall protective effect against ischemic injury as determined by the release of enzymes like creatine kinase and LDH. We did not note inflammatory or immune responses elicited by the expression of hsps in transgenic animals and cardiac myocytes. The specific cell types in which hsps are expressed may contribute to the protective effect of hsps versus their inflammatory and immunogenic effects when expressed in other cell types. Infect. Dis. Obstet. Gynecol. 7:55–57, 1999.

An experimental approach to production of peptide-like compounds in the early terrestrial planets

A mixture of six DL-amino acids in prebiotic proportions were immersed in heavy mineral oil, warmed up to 240°C and exposed to the action of spark discharges on a mixture of H 2O and CO 2 gases. Water soluble products were analyzed by Fourier transformed infrared spectroscopy (FTIR) showing yields of peptide-like compounds characterized by amidine linkages and other molecules consisting of aromatic and non-aromatic alcohols and nitrogenous aromatics. Non-soluble products analyzed by scanning electron microscopy (SEM) revealed the presence of a material comparable to tholins whereas the products soluble in water showed the presence of an amphiphilic component. The recovered amounts and the FTIR spectra corresponding to heavy mineral oil exposed to simulated experimental conditions including UVC-radiations, clearly show no difference relative to initial sample indicating that the decomposition rate of these molecules should be very slow at the time. Therefore, we consider plausible that alkane environments ⩾ n-C 18 occurring on the early terrestrial planets could offer several advantages for the synthesis and survival of nitrogenous molecules, particularly towards the end or after the age of the large impacts, such as protection against oxidizing atmospheres, high temperatures generated by greenhouse effects or shock waves and protection against decomposition effects caused by high UVC-radiations, corona discharges and lightning. Currently, these facts could be happening in other regions of our solar system and in extrasolar planets allowing the formation and accumulation of complex nitrogenous molecules.

Performance Metrics for Composite Integral Armor

Future combat systems necessarily focus on lightweight, highly mobile and transportable armored vehicles. Lightweight composite integral armor systems are being developed to meet these needs. The goal of this paper is to centrally document the myriad design requirements for composite integral armors that serve multifunctional roles including ballistic, structural, shock, electromagnetic, and fire protection. Structural and ballistic performance requirements as well as manufacturing and life-cycle performance issues of integral armor are presented. Specific areas addressed include high-strain-rate testing and modeling, ballistic testing and modeling, low-cycle fatigue, damage tolerance, repair, reduced-step processing, through-thickness reinforcement, energy dissipation and rate-dependent failure mechanisms, and non-linear mechanics.