Sample records for bio dosemeters hsp

  1. HSP as bio dosemeters; HSP como biodosimetros

    Energy Technology Data Exchange (ETDEWEB)

    Manzanares A, E.; Vega C, H.R. [Grupo Academico de Radiobiologia, UAZ, A.P. 336, 98000 Zacatecas (Mexico)


    A review of the more important passages for the field of the radiological protection, from the discovery, beginnings of the utilization of the radiations emitting, passing to the legislation proposals and the transformations that had been suffer to establish the international standards of radiological protection. The paradigms on the effects and chronologically the emanated discoveries of the cellular research and the measurement of cellular products in the species and the diverse alive beings making emphasis in evaluating the expression of the heat-shock proteins caused by low doses of gamma radiation, these are used as radiation bio monitors and of other agents that cause cellular stress and lastly a prospective of the future of the radiobiology. (Author)

  2. Review of international standards for dosemeters. (United States)

    Behrens, R; Ambrosi, P


    International standards for radiation protection dosemeters are published by the International Electrotechnical Commission and the International Organization for Standardization. Several standards exist side by side, although they treat the same measuring task, and specify different requirements, so that dosemeters of different quality result. In this paper, the quality of dosemeters is compared by calculating the uncertainty of dose measurements for dosemeters, which just basely fulfil the respective standard. The results are related to general yardsticks on uncertainty laid down by international organisations. Furthermore, technical differences are standards and addressed and a method to make them conform is presented.

  3. Passive neutron dosemeter with activation detector

    Energy Technology Data Exchange (ETDEWEB)

    Valero L, C.; Banuelos F, A.; Guzman G, K. A.; Borja H, C. G.; Hernandez D, V. M.; Vega C, H. R., E-mail: [Universidad Autonoma de Zacatecas, Unidad Academica de Estudios Nucleares, Calle Cipres No. 10, Fracc. La Penuela, 98068 Zacatecas (Mexico)


    A passive neutron dosemeter with {sup 197}Au activation detector has been developed. The area dosemeter was made as a 20.5 {phi} x 20.5 cm{sup 2} polyethylene moderator, with a polyethylene pug where a {sup 197}Au foil can be located either parallel or perpendicular to moderator axis. Using Monte Carlo methods, with the MCNP5 code. With the fluence response and the fluence-to-equivalent dose conversion coefficients from ICRP-74, responses to H*(10) were also calculated, these were compared against responses of commercially available neutron area monitors and dosemeters. (Author)

  4. The relative merits of discriminating and non-discriminating dosemeters

    DEFF Research Database (Denmark)

    Marshal, T. O.; Christensen, Palle; Julius, H. W.;


    The need for discriminating and non-discriminating personal dosemeters in the field of radiological protection is examined. The ability of various types of dosemeter to meet these needs is also discussed. It is concluded that there is a need for discriminating dosemeters but in the majority of ca...

  5. Colour dosemeters for high level radiation dosimetry

    DEFF Research Database (Denmark)

    Schönbacher, H.; Coninckx, F.; Miller, A.;


    interpretation or a dose measurement with a simple instrument such as a portable reflecting densitometer in the range of 10(3) to 10(6) Gy. Two projects were investigated: (1) a thin plastic film with a self adhesive tape containing a radiochromic dye which induces a colour change when exposed to ionising...... radiation; and (2) a paint containing a base substance with a pigment. The paint dosemeter remained unaffected by irradiation up to 3 x 10(4) Gy while the film dosemeter showed a measurable colour change from 10(4) Gy to 5 x 10(5) Gy. Above 10(6) Gy the film dosemeter is destroyed by radiation. Samples...

  6. Comparison of main thermoluminescent properties of some TL dosemeters

    DEFF Research Database (Denmark)

    Prokic, M.; Bøtter-Jensen, L.


    Different types of TL dosemeters that are attractive for dosimetry practice were investigated and compared. The aim of this study was to evaluate the TL characteristics of different TL dosemeters using two basically different types of commercially available TL readers. The findings demonstrate...

  7. The increasing need for active dosemeters in the nuclear industry

    Energy Technology Data Exchange (ETDEWEB)

    Hirning, C.R. [Ontario Hydro, Whitby, ON (Canada). Health Physics Dept.


    As a legal and administrative dose limits come down, there is an increasing need in the nuclear industry for greater accuracy in dose measurement, for lower detection limits, and for more rapid reporting of doses, Active (i.e. electronic) dosemeters offer the possibility of substantial improvements in these areas. The advantages of electronic dosemeters are reviewed, some impediments to their more rapid adoption considered, some needs for research and development suggested, and the implications of a change from passive to active dosemeters looked at. A review of the present status of electronic dosemeter testing, standard-setting and system implementation is given. The next major era in the history of radiation dosimetry appears to be well under way. (Author).

  8. Response of Alanine Dosemeter to Heavy Ions

    Institute of Scientific and Technical Information of China (English)

    LiWenjian; SuXu; YangYingjie; YuanJianlei; DangBingrong; WangXiao; MaQiufeng; ZhouLibin; HaoJifang; MaoShuhong


    The amino acid L-α-alanine has been investigated for use as a radiation detector in low and high LET radiation fields[1]. The radiatioa detector is cheap and easy to handle. The radiation inducing free radicals are stable at normal laboratory conditions for doses below 104 Gy over a long period of time, which makes the detector useful for intercomparison and documentation purposes. The dosimetric features of alanine-based electron spin resonance (ESR) detectors in high energy electron beams used in radiotherapy were considered[2]. The 5 mm long alanine detectors were found to be the most suitable for carrying out in vivo dosimetry on patients undergoing electron beam radiotherapy. However, data concerning dosimetry of the alanine dosemeter to heavy charged particles are lacking, especially in China.

  9. Prediction analysis of dose equivalent responses of neutron dosemeters used at a MOX fuel facility. (United States)

    Tsujimura, N; Yoshida, T; Takada, C


    To predict how accurately neutron dosemeters can measure the neutron dose equivalent (rate) in MOX fuel fabrication facility work environments, the dose equivalent responses of neutron dosemeters were calculated by the spectral folding method. The dosemeters selected included two types of personal dosemeter, namely a thermoluminescent albedo neutron dosemeter and an electronic neutron dosemeter, three moderator-based neutron survey meters, and one special instrument called an H(p)(10) monitor. The calculations revealed the energy dependences of the responses expected within the entire range of neutron spectral variations observed in neutron fields at workplaces.

  10. Potential clinical utility of a fibre optic-coupled dosemeter for dose measurements in diagnostic radiology. (United States)

    Jones, A Kyle; Hintenlang, David


    Many types of dosemeters have been investigated for absorbed dose measurements in diagnostic radiology, including ionisation chambers, metal-oxide semiconductor field-effect transistor dosemeters, thermoluminescent dosemeters, optically stimulated luminescence detectors, film and diodes. Each of the aforementioned dosemeters suffers from a critical limitation, either the need to interrogate, or read, the dosemeter to retrieve dose information or large size to achieve adequate sensitivity. This work presents an evaluation of a fibre optic-coupled dosemeter (FOCD) for use in diagnostic radiology dose measurement. This dosemeter is small, tissue-equivalent and capable of providing true real-time dose information. The FOCD has been evaluated for dose linearity, angular dependence, sensitivity and energy dependence at energies, beam qualities and beam quantities relevant to diagnostic radiology. The FOCD displayed excellent dose linearity and high sensitivity, while exhibiting minimal angular dependence of response. However, the dosemeter does exhibit positive energy dependence, and is subject to attenuation of response when bent.

  11. Graphite mixed magnesium borate TL dosemeters for beta ray dosimetry

    DEFF Research Database (Denmark)

    Prokic, M; Christensen, Poul


    Sintered MgB4O7:Dy dosemeters with graphite contents from 1 to 10% were investigated for application for personnel dosimetry. Data are given on dose response, dose threshold, reproducibility, beta energy response and fading. Furthermore, results from practical field experiments are presented...

  12. Radiological environmental monitoring with LiF-700-H dosemeters

    Energy Technology Data Exchange (ETDEWEB)

    Scarnichia, E.; Levanon, I.; Andres, P.; Miani, C.; Ramirez, S., E-mail: [Comision Nacional de Energia Atomica, Centro Atomico Bariloche, Grupo Proteccion Radiologica, Exequiel Bustillo 5AV 9500, R8402AGP San Carlos de Bariloche, Rio Negro (Argentina)


    Since 2008 a radiological environmental monitoring with LiF-700-H has been carried out as a result of increasing the Ra-6 research reactor core power. The information obtained is used to evaluate and to quantify analytically the air kerma rate, the fading and the associated uncertainty by developing software tools (deconvolution and uncertainty algorithms). LiF-700-H dosemeters were chosen because of their high sensitivity to low air kerma rates. They show a very good stability and a negligible fading for two-month working periods. The air kerma rate detection limit (based on the 3{sigma} criterion) during these working periods is about 0.4 n Gy/h. Air kerma rates of about 70 n Gy/h are measured with this detection limit. Following the Nist guidelines, an algorithm was developed in order to find the associated uncertainty. It considers several aspects, such as the source activity decay, distance source-dosemeter during the calibration procedure, irradiation time, calibration factor, dosemeter readout, dosemeter sensitivity, TLD reader stability and fading. The associated uncertainty is found to be about 25% for a 95% confidence interval (k = 2.025), which can be considered acceptable when taking into account the very low air kerma rates estimated. The LiF-700-H response to different energies and its relationship with climate changes over the calendar year are planned as future tasks. (Author)

  13. Household and workplace chemicals as retrospective luminescence dosemeters

    DEFF Research Database (Denmark)

    Thomsen, Kristina Jørkov; Bøtter-Jensen, L.; Murray, A.S.


    In the development of techniques for the retrospective assessment of the close absorbed by communities living and working adjacent to the site of a nuclear accident, attention has concentrated on the use Of natural minerals such as quartz and feldspar as dosemeters. These minerals are widely found...

  14. Evaluation of BICRON NE MCP DXT-RAD passive extremity dosemeter

    CERN Document Server

    Yuen, P S; Frketich, G; Rotunda, J


    Passive extremity dosemeters currently used in dosimetry communities worldwide have shortcomings. In general, an extremity dosemeter has too thick a detector element, and the dosemeter response is highly energy dependent for beta rays with energies ranging from 200 keV to 2 MeV. It often does not have dosemeter identification, causing problems in the chain of custody. It is often read manually, rendering reading/packing operations very labour intensive. As a result of collaboration between AECL and BICRON NE, a new extremity dosemeter, incorporating a highly sensitive LiF:Mg,Cu,P TLD and tentatively code named MCP DXT-RAD, was developed. It has been evaluated for radiological performance against an ISO draft standard for extremity dosemeters in twelve categories: homogeneity, detection threshold, beta ray energy response, beta angular response, photon energy response, photon angular response, reproducibility, stability under various climatic conditions, linearity, residue, self irradiation, and effect of ligh...

  15. Photon and fast neutron dosimetry using aluminium oxide thermoluminescence dosemeters. (United States)

    Santos, J P; Fernandes, A C; Gonçalves, I C; Marques, J G; Carvalho, A F; Santos, L; Cardoso, J; Osvay, M


    Al(2)O(3):Mg,Y thermoluminescence (TL) dosemeters were used to measure photon and fast neutron doses in a fast neutron beam recently implemented at the Portuguese Research Reactor, Nuclear and Technological Institute, Portugal. The activation of Al(2)O(3):Mg,Y by fast neutrons provides information about the fast neutron component by measuring the activity of the reaction products and the self-induced TL signal. Additionally, the first TL reading after irradiation determines the photon dose. The elemental composition of the dosemeters was determined by instrumental neutron activation analysis and by particle induced X-ray emission. Results demonstrate that Al(2)O(3):Mg,Y is an adequate material to discriminate photon and fast neutron fields for reactor dosimetry purposes.

  16. Theoretical aspects of the design of a passive radon dosemeter. (United States)

    Wilkinson, P; Saunders, B J


    Some mathematical aspects of the development and design of a passive radon dosemeter are considered. In particular, a mathematical model is presented that is concerned with the gaseous diffusion of radon into a confined region bounded by a plastic material of known diffusion coefficient. The relationship between the time-integrated radon concentrations, inside and outside a sealed plastic container are derived. Estimates of the exposure of people to radon can be made using the time integrated radon concentration inside a calibrated container containing a CR-39 etched-track device. As a consequence of the analysis, it is possible to design a passive radon dosemeter that will be accurate, resistant to moisture and whose response will be independent of rapid variations in radon concentration. The possibility of using a container of this type for the measurement of diffusion coefficients is discussed.

  17. Development of active environmental and personal neutron dosemeters. (United States)

    Nakamura, T; Nunomiya, T; Sasaki, M


    For neutron dosimetry in the radiation environment surrounding nuclear facilities, two types of environmental neutron dosemeters, the high-sensitivity rem counter and the high-sensitivity multi-moderator, the so-called Bonner ball, have been developed and the former is commercially available from Fuji Electric Co. By using these detectors, the cosmic ray neutrons at sea level have been sequentially measured for about 3 y to investigate the time variation of neutron spectrum and ambient dose equivalent influenced by cosmic and terrestrial effects. Our Bonner ball has also been selected as the neutron detector in the International Space Station and has already been used to measure neutrons in the US experimental module. The real time wide-range personal neutron dosemeter which uses two silicon semiconductor detectors has been developed for personal dosimetry and is commercially available from Fuji Electric Co. This dosemeter has good characteristics, fitted to the fluence-to-dose conversion factor in the energy range from thermal energies to several tens of mega-electron-volts and is now widely used in various nuclear facilities.

  18. Peptide substrate identification for yeast Hsp40 Ydj1 by screening the phage display library

    Directory of Open Access Journals (Sweden)

    Li Jingzhi


    Full Text Available We have identified a peptide substrate for molecular chaperone Hsp40 Ydj1 by utilizing the combination of phage display library screening and isothemol titration calirimetry (ITC. The initial peptide substrate screening for Hsp40 Ydj1 has been carried out by utilizing a 7-mer phage display library. The peptide sequences from the bio-panning were synthesized and object to the direct affinity measurement for Hsp40 Ydj1 by isothemol titration calirimetry studies. The peptide which has the measurable affinity with Ydj1 shows enriched hydrophobic residues in the middle of the substrate fragment. The peptide substrate specificity for molecular chaperone Hsp40 has been analyzed.

  19. New concept of IEC standards for radiation protection dosemeters. (United States)

    Ambrosi, P; Behrens, R


    The International Electrotechnical Commission (IEC) develops new standards for radiation protection dosemeters which follow a new concept. They are much more flexible in detail, but still ensure the same measurement quality. They are, for example, no longer specific for the detector type, but rather specific for the measurement task, e.g. for individual monitoring with active direct-reading instruments. Another example is that they are flexible with respect to the ranges of influence quantities. The conceptual changes are described in this paper, together with the advantages this new concept provides for manufacturers, users and legislators.

  20. Type testing of the Siemens Plessey electronic personal dosemeter

    Energy Technology Data Exchange (ETDEWEB)

    Hirning, C.R. [Ontario Hydro, Health Physics Dept., Whitby, Ontario (Canada); Yuen, P.S


    This paper presents the results of a laboratory assessment of the performance of a new type of personal dosimeter, the Electronic Personal Dosemeter made by Siemens Plessey Controls Limited. Twenty pre-production dosimeters and a reader were purchased by Ontario Hydro for the assessment. Tests were performed on radiological performance, including reproducibility, accuracy, linearity, detection threshold, energy response, angular response, neutron response, and response time. There were also tests on the effects of a variety of environmental factors, such as temperature, humidity, pulsed magnetic and electric fields, low- and high-frequency electromagnetic fields, light exposure, drop impact, vibration, and splashing. Other characteristics that were tested were alarm volume, clip force, and battery life. The test results were compared with the relevant requirements of three standards: an Ontario Hydro standard for personal alarming dosimeters, an International Electrotechnical Commission draft standard for direct reading personal dose monitors, and an International Electrotechnical Commission standard for thermoluminescence dosimetry systems for personal monitoring. In general, the performance of the Electronic Personal Dosemeter was found to be quite acceptable: it met most of the relevant requirements of the three standards. However, the following deficiencies were found: slow response time; sensitivity to high-frequency electromagnetic fields; poor resistance to dropping and an alarm that was not loud enough. In addition, the response of the electronic personal dosimeter to low-energy beta rays may be too low for some applications. Problems were experienced with the reliability of operation of the pre-production dosimeters used in these tests. (author)

  1. Tupe testing of the Siemens Plessey electronic personal dosemeter

    Energy Technology Data Exchange (ETDEWEB)

    Hirning, C.R. [Ontario Hydro, Whitby, Ontario (Canada); Yuen, P.S. [Chalk River Laboratories, Chalk River, Ontario (Canada)


    This paper presents the results of a laboratory assessment of the performance of a new type of personal dosimeter, the Electronic Personal Dosemeter made by Siemens Plessey Controls Limited. Twenty pre-production dosimeters and a reader were purchased by Ontario Hydro for the assessment. Tests were performed on radiological performance, including reproducibility, accuracy, linearity, detection threshold, energy response, angular response, neutron response, and response time. There were also tests on the effects of a variety of environmental factors, such as temperature, humidity, pulsed magnetic and electric fields, low- and high-frequency electromagnetic fields, light exposure, drop impact, vibration, and splashing. Other characteristics that were tested were compared with the relevant requirements of three standards: an Ontario Hydro standard for personal alarming dosimeters, an International Electrotechnical Commission draft standard for direct reading personal dose monitors, and an International Electrotechnical Commission standard for thermoluminescence dosimetry systems for personal monitoring. In general, the performance of the Electronic Personal Dosemeter was found to be quite acceptable: it met most of the relevant requirements of the three standards. However, the following deficiencies were found: slow response time; sensitivity to high-frequency electromagnetic fields; poor resistance to dropping; and an alarm that was not loud enough. In addition, the response of the electronic personal dosimeter to low-energy beta rays may be too low for some applications. Problems were experienced with the reliability of operation of the pre-production dosimeters used in these tests. 11 refs., 8 figs., 24 tabs.

  2. Type testing of the Siemens Plessey electronic personal dosemeter. (United States)

    Hirning, C R; Yuen, P S


    This paper presents the results of a laboratory assessment of the performance of a new type of personal dosimeter, the Electronic Personal Dosemeter made by Siemens Plessey Controls Limited. Twenty pre-production dosimeters and a reader were purchased by Ontario Hydro for the assessment. Tests were performed on radiological performance, including reproducibility, accuracy, linearity, detection threshold, energy response, angular response, neutron response, and response time. There were also tests on the effects of a variety of environmental factors, such as temperature, humidity, pulsed magnetic and electric fields, low- and high-frequency electromagnetic fields, light exposure, drop impact, vibration, and splashing. Other characteristics that were tested were alarm volume, clip force, and battery life. The test results were compared with the relevant requirements of three standards: an Ontario Hydro standard for personal alarming dosimeters, an International Electrotechnical Commission draft standard for direct reading personal dose monitors, and an International Electrotechnical Commission standard for thermoluminescence dosimetry systems for personal monitoring. In general, the performance of the Electronic Personal Dosemeter was found to be quite acceptable: it met most of the relevant requirements of the three standards. However, the following deficiencies were found: slow response time; sensitivity to high-frequency electromagnetic fields; poor resistance to dropping; and an alarm that was not loud enough. In addition, the response of the electronic personal dosimeter to low-energy beta rays may be too low for some applications. Problems were experienced with the reliability of operation of the pre-production dosimeters used in these tests.

  3. Hsp104 and prion propagation. (United States)

    Romanova, Nina V; Chernoff, Yury O


    High-ordered aggregates (amyloids) may disrupt cell functions, cause toxicity at certain conditions and provide a basis for self-perpetuated, protein-based infectious heritable agents (prions). Heat shock proteins acting as molecular chaperones counteract protein aggregation and influence amyloid propagation. The yeast Hsp104/Hsp70/Hsp40 chaperone complex plays a crucial role in interactions with both ordered and unordered aggregates. The main focus of this review will be on the Hsp104 chaperone, a molecular "disaggregase".

  4. Dosimetric characteristics of a TLD dosemeter with extremities; Caracteristicas dosimetricas de un dosimetro TLD de extremidades

    Energy Technology Data Exchange (ETDEWEB)

    Molina P, D.; Diaz B, E.; Lien V, R. [Centro de Proteccion e Higiene de las Radiaciones, CPHR, Apdo.Postal 6195, Habana 6, CP 10600, Ciudad Habana (Cuba)


    It was designed a TLD dosemeter for the monitoring of the extremities. This one consists in a metallic ring with a circular orifice where is arranged a T L detector of LiF: Mg,Ti (Model JR1152C) 5 x 5 x 0.8 mm{sup 3} covered by a polyethylene fine layer. In this work were studied the dosimetric properties of the dosemeter for its application in the dosimetry of extremities for photonic radiation. the results obtained allow conclude that the designed dosemeter can be used for the extremities monitoring. (Author)

  5. Type testing of personal dosemeters for photon energy and angular response

    DEFF Research Database (Denmark)

    Julius, H.W.; Marshall, T.O.; Christensen, Poul;


    Performance criteria for personal dosemeters have been recently proposed in the CEC document 'Technical Recommendations for Monitoring Individuals Occupationally Exposed to External Radiation' (EUR 5287, Revised). The current experiment was carried out to demonstrate that TLD-based dosimetry...

  6. Determination of relevant parameters for the use of electronic dosemeters in pulsed fields of ionising radiation. (United States)

    Zutz, H; Hupe, O; Ambrosi, P; Klammer, J


    Active electronic dosemeters using counting techniques are used for radioprotection purposes in pulsed radiation fields in X-ray diagnostics or therapy. The disadvantage of the limited maximum measurable dose rate becomes significant in these radiation fields and leads to some negative effects. In this study, a set of relevant parameters for a dosemeter is described, which can be used to decide whether it is applicable in a given radiation field or not. The determination of these relevant parameters-maximum measurable dose rate in the radiation pulse, dead time of the dosemeter, indication per counting event and measurement cycle time-is specified. The results of the first measurements on the determination of these parameters for an electronic personal dosemeter of the type Thermo Fisher Scientific EPD Mk2 are shown.

  7. Sensitivity of a LR-115 based radon dosemeter

    CERN Document Server

    Bagnoli, F; Bucci, S


    The first results of a study on the sensitivity of a LR-115 based radon dosemeter as a function of the absorber thickness are presented. The theoretical sensitivity was analytically calculated considering a constant detector response to alpha particles within a given energy range and up to a critical angle of incidence. The results are presented in two extreme situations: i) both radon and its decay products uniformly distributed in the chamber volume; ii) radon decay products uniformly deposited on the chamber walls. The agreement with the experimental curve shape appears better in the former case, suggesting that either the parameter values of the model could be different from the chosen values, or the model was too simplified.

  8. 1983 ORNL intercomparison of personnel neutron and gamma dosemeters

    Energy Technology Data Exchange (ETDEWEB)

    Swaja, R.E.; Sims, C.S.; Greene, R.T.


    The Ninth Personnel Dosimetry Intercomparison Study was conducted during April 19-21, 1983, at the Oak Ridge National Laboratory. Dosemeters from 33 participating agencies were mounted on water-filled polyethylene elliptical phantoms and exposed to a range of low-level dose equivalents (0.02-0.45 mSv gamma and 0.49-11.14 mSv neutron) which could be encountered during routine personnel monitoring in mixed radiation fields. The Health Physics Research Reactor served as the radiation source for six separate exposures which used four different shield conditions: unshielded and shielded with steel, steel/concrete, and concrete. Results of the neutron measurements indicate that it is not unusual for dose equivalent estimates made under the same conditions by different agencies to differ by more than a factor of 2. Albedo systems, which were the most popular neutron monitors in this study, provided the most accurate results with CR-39 recoil track being least accurate. Track and film neutron systems exhibited problems providing measurable indication of neutron exposure at dose equivalents of about 0.50 mSv. Gamma measurements showed that TLD and film systems generally overestimated dose equivalents in the mixed radiation fields with film exhibiting significant problems providing measurable indication of gamma exposure at dose equivalents lower than about 0.15 mSv. Under the conditions of this study in which exposures were carefully controlled and participants had information concerning exposure conditions and incident spectra prior to dosemeter analysis, only slightly more than half of all neutron and gamma dose equivalent estimates met regulatory accuracy standards relative to reference values. These results indicate that continued improvement of mixed-field personnel dosimetry is required by many participating organizations. 15 references, 30 tables.

  9. Mechanisms of HSP72 release

    Indian Academy of Sciences (India)

    Alexzander Asea


    Currently two mechanisms are recognized by which heat shock proteins (HSP) are released from cells; a passive release mechanism, including necrotic cell death, severe blunt trauma, surgery and following infection with lytic viruses, and an active release mechanism which involves the non classical protein release pathway. HSPs are released both as free HSP and within exosomes. This review covers recent findings on the mechanism by which stress induces the release of HSP72 into the circulation and the biological significance of circulating HSP72 to host defense against disease.

  10. Hsp40s specify functions of Hsp104 and Hsp90 protein chaperone machines.

    Directory of Open Access Journals (Sweden)

    Michael Reidy


    Full Text Available Hsp100 family chaperones of microorganisms and plants cooperate with the Hsp70/Hsp40/NEF system to resolubilize and reactivate stress-denatured proteins. In yeast this machinery also promotes propagation of prions by fragmenting prion polymers. We previously showed the bacterial Hsp100 machinery cooperates with the yeast Hsp40 Ydj1 to support yeast thermotolerance and with the yeast Hsp40 Sis1 to propagate [PSI+] prions. Here we find these Hsp40s similarly directed specific activities of the yeast Hsp104-based machinery. By assessing the ability of Ydj1-Sis1 hybrid proteins to complement Ydj1 and Sis1 functions we show their C-terminal substrate-binding domains determined distinctions in these and other cellular functions of Ydj1 and Sis1. We find propagation of [URE3] prions was acutely sensitive to alterations in Sis1 activity, while that of [PIN+] prions was less sensitive than [URE3], but more sensitive than [PSI+]. These findings support the ideas that overexpressing Ydj1 cures [URE3] by competing with Sis1 for interaction with the Hsp104-based disaggregation machine, and that different prions rely differently on activity of this machinery, which can explain the various ways they respond to alterations in chaperone function.

  11. Numerical and experimental results of the operational neutron dosemeter 'Saphydose-N'. (United States)

    Lahaye, T; Chau, Q; Ménard, S; Ndontchueng-Moyo, M; Bolognese-Milsztajn, T; Rannou, A


    Since 1993, the Institute for Radiological Protection and Nuclear Safety (IRSN) has lead, in association with Electricité de France (EDF), a R&D study of a neutron personal electronic dosemeter. This dosemeter, called 'Saphydose-N', is manufactured by the SAPHYMO company. This paper presents first the optimisation of some detector components using Monte Carlo calculations, and second the test of the manufactured Saphydose-N under radiation following the IEC 1323 standard's recommendations for active personal neutron dosemeters. The measurements with the manufactured dosemeter were performed on the one hand at PTB (Physikalisch-Technische Bundesanstalt) in mono-energetic neutron fields and, on the other hand at IRSN in neutron fields generated by a thermal facility (SIGMA), radionuclide ISO sources and a realistic spectrum (CANEL/T400). The manufactured dosemeter Saphydose-N was also tested during measurement campaigns of the European programme EVIDOS ('Evaluation of Individual Dosimetry in Mixed Neutron and Photon Radiation Fields') at different nuclear workplaces. The study showed that Saphydose-N complies with the recommendations of standard IEC 1323 and can be used at any workplace with no previous knowledge of the neutron field characteristics.

  12. Eurados trial performance test for personal dosemeters for external beta radiation

    DEFF Research Database (Denmark)

    Christensen, P.; Bordy, J.M.; Ambrosi, P.


    was accomplished, This paper deals with the part of the performance test concerned with exposure to beta radiation. Fifteen dosimetric services participated with whole-body dosemeters intended to measure beta doses (H-p(0.07)) of which 13 used thermoluminescent (TL) detectors and two used photographic films. Eight......On the initiative of the European Dosimetry Group (EURADOS) action group 'Harmonisation and Dosimetric Quality Assurance in Individual Monitoring for External Radiation' a trial performance test for whole-body and extremity personal dosemeters broadly representative of those in use in the EU...... the results obtained from the exercise. In particular, based on the replies to a questionnaire issued to each participant, the results are analysed in relation to important design characteristics of the dosemeters taking part in the test....

  13. Independent evaluation of optically stimulated luminescence (OSL) 'dot' dosemeters for environmental monitoring. (United States)

    Timilsina, Bindu; Gesell, Thomas F


    Optically stimulated luminescence (OSL) 'dot' dosemeters (manufactured by Landauer®) are reported to have a high degree of environmental stability, high level of sensitivity and provide wide range of dose measuring capabilities from 0.05 mGy to 100 Gy. The optical read out method is fast and relatively simple and permits repeated read out, but few studies have been performed about its application in monitoring radiation in the environment. This study was initiated to independently test the performance of OSL dot dosemeters for the application of measuring doses of radiation in the outdoor environment. Testing was performed in the laboratory to evaluate reproducibility and stability and in the field to evaluate accuracy relative to calibrated high-pressure ionisation chambers. The results showed that OSL dot dosemeters had good reproducibility and stability in both laboratory and field tests and met the performance requirements of standards of the American National Standards Institute.

  14. Mixed field personnel dosimetry using a nearly tissue-equivalent multi-element thermoluminescence dosemeter

    Energy Technology Data Exchange (ETDEWEB)

    Moscovitch, M.; Velbeck, K.J.; Bencke, G.M. (Engelhard Corp., Harshaw Crystal and Electronic Products, Solon, OH (USA))


    The method of response interpretation for determining dose equivalents of a new multi-element thermoluminescence dosemeter in mixed photon-beta and photon-neutron fields is described. The four TL chips are LiF:Mg,Ti, each covered by its own unique filter. Experiments to determine the response of the dosemeter and to test the dose calculation algorithm were performed according to the standard established by the US Department of Energy Laboratory Accreditation Program (DOELAP). It has been shown that the dosemeter response can be used to identify the mixture type. The accuracy of the system has been proven in blind testing to be well within DOELAP tolerance limits in all test categories. (author).

  15. Theoretical study and calculation of the response of a fast neutron dosemeter based on track detection

    Energy Technology Data Exchange (ETDEWEB)

    Decossas, J.L.; Vareille, J.C.; Moliton, J.P.; Teyssier, J.L. (Limoges Univ., 87 (France). Lab. d' Electronique des Polymeres sous Faisceaux Ioniques)


    A fast neutron dosemeter is generally composed of a radiator in which n-p elastic scattering occurs and a detector which registers protons. A theoretical study, and the calculation (FORTRAN program) of the response of such a dosemeter is presented involving two steps: 1) The proton flux emerging from a thick radiator on which monoenergetic neutrons are normally incident is studied. This is characterised by its energy spectrum depending on the neutron energy and on the radiator thickness. 2) Proton detection being achieved with a solid state nuclear track detector whose performance is known, the number of registered tracks are calculated. The dosemeter sensitivity (tracks cm/sup -2/. Sv/sup -1/) is deduced. Then, the calculations show that it is possible to optimise the radiator thickness to obtain the smallest variation in sensitivity with neutron energy. The theoretical results are in good agreement with the experimental ones found in the literature.

  16. Elimination of Cerenkov interference in a fibre-optic-coupled radiation dosemeter. (United States)

    Justus, Brian L; Falkenstein, Paul; Huston, Alan L; Plazas, Maria C; Ning, Holly; Miller, Robert W


    An optical fibre point dosemeter based on the gated detection of the luminescence from a Cu(1+)-doped fused quartz detector effectively eliminated errors due to Cerenkov radiation and native fibre fluorescence. The gated optical fibre dosemeter overcomes serious problems faced by scintillation and optically stimulated luminescence approaches to optical fibre point dosimetry. The dosemeter was tested using an external beam radiotherapy machine that provided pulses of 6 MV X rays. Gated detection was used to discriminate the signal collected during the radiation pulses, which included contributions from Cerenkov radiation and native fibre fluorescence, from the signal collected between the radiation pulses, which contained only the long-lived luminescence from the Cu(1+)-doped fused quartz detector. Gated detection of the luminescence provided accurate, real-time dose measurements that were linear with absorbed dose, independent of dose rate and that were accurate for all field sizes studied.


    Sokolov, Alexey; Fehrenbacher, Georg; Radon, Torsten


    For the radiation survey at intermediate and high-energy accelerators, there is a need for a neutron dosemeter which provides reliable readings of the neutron dose in a wide energy range for continuous and pulsed radiation. The objective of this development is to find a dosemeter that fulfils the necessary requirements and can be reliably used to prove that the radiation levels in areas around accelerators are in accordance with the limits of the respective radiation protection legislation. A simple layout with small dimensions and light weight as well as the usage of common materials to lower the production costs is to be achieved.

  18. The assembly and intermolecular properties of the Hsp70-Tomm34-Hsp90 molecular chaperone complex. (United States)

    Trcka, Filip; Durech, Michal; Man, Petr; Hernychova, Lenka; Muller, Petr; Vojtesek, Borivoj


    Maintenance of protein homeostasis by molecular chaperones Hsp70 and Hsp90 requires their spatial and functional coordination. The cooperation of Hsp70 and Hsp90 is influenced by their interaction with the network of co-chaperone proteins, some of which contain tetratricopeptide repeat (TPR) domains. Critical to these interactions are TPR domains that target co-chaperone binding to the EEVD-COOH motif that terminates Hsp70/Hsp90. Recently, the two-TPR domain-containing protein, Tomm34, was reported to bind both Hsp70 and Hsp90. Here we characterize the structural basis of Tomm34-Hsp70/Hsp90 interactions. Using multiple methods, including pull-down assays, fluorescence polarization, hydrogen/deuterium exchange, and site-directed mutagenesis, we defined the binding activities and specificities of Tomm34 TPR domains toward Hsp70 and Hsp90. We found that Tomm34 TPR1 domain specifically binds Hsp70. This interaction is partly mediated by a non-canonical TPR1 two-carboxylate clamp and is strengthened by so far unidentified additional intermolecular contacts. The two-carboxylate clamp of the isolated TPR2 domain has affinity for both chaperones, but as part of the full-length Tomm34 protein, the TPR2 domain binds specifically Hsp90. These binding properties of Tomm34 TPR domains thus enable simultaneous binding of Hsp70 and Hsp90. Importantly, we provide evidence for the existence of an Hsp70-Tomm34-Hsp90 tripartite complex. In addition, we defined the basic conformational demands of the Tomm34-Hsp90 interaction. These results suggest that Tomm34 represents a novel scaffolding co-chaperone of Hsp70 and Hsp90, which may facilitate Hsp70/Hsp90 cooperation during protein folding.

  19. Testing the direct ion storage dosemeter for personal dosimetry in a nuclear research centre and a hospital. (United States)

    Vanhavere, F; Covens, P


    The direct ion storage (DIS) dosemeter can have some clear advantages in personal dosimetry. Before introducing the DIS into practice in the dosimetry service, a series of tests was performed on the linearity, angular and energy dependence, temperature influences and hard resets. After that, for several months, the DIS dosemeters were worn in parallel with the legal dosemeters (thermoluminescent badge) in a nuclear research centre and in several departments of a university hospital. The conclusions are that the DIS has good characteristics to be used as legal personal dosemeter, and that the comparison with the TLD badge is good. Only in interventional radiology and cardiology fields the DIS gives significant lower values than the TLD badge.


    Cardoso, J; Santos, L; Carvalhal, G; Oliveira, C


    The calibration of electronic personal dosemeters at the Portuguese ionizing radiation metrology laboratory uses the standard IEC 61526 for calibration methodology. This standard describes the irradiation geometry for testing and indicates that the standard ISO 4037-1, 2, 3 and 4 should be used. The ISO 4037 establishes that the reference point of test is a point in the radiation monitor, known or established, and the calibration phantom should be placed on its back in order to simulate the trunk body. Recently, ISO published another standard, the ISO 29661, that changes the reference point from the radiation monitor to the front face of the calibration phantom. The aim of this work is to present the result of the comparison of these two methodologies on personal dosemeters from five different manufacturers. The work shows differences in the Hp(10) response up to 4% resulting from the two different reference point concepts.

  1. Measurement models for passive dosemeters in view of uncertainty evaluation using the Monte Carlo method. (United States)

    van Dijk, J W E


    Two measurement models for passive dosemeters such as thermoluminescent dosemeter, optically stimulated luminescence, radio-photoluminescence, photographic film or track etch are discussed. The first model considers the dose evaluation with the reading equipment as a single measurement, the one-stage model. The second model considers the build-up of a latent signal or latent image in the detector during exposure and the evaluation using a reader system as two separate measurements, the two-stage model. It is discussed that the two-stage model better reflects the cause and effect relations and the course of events in the daily practice of a routine dosimetry service. The one-stage model will be non-linear in crucial input quantities which can give rise to erroneous behavior of the uncertainty evaluation based on the law of propagation of uncertainty. Input quantities that show an asymmetric probability distributions propagate through the one-stage model in a physically not relevant way.

  2. Application of TL dosemeters for dose distribution measurements at high temperatures in nuclear reactors. (United States)

    Osvay, M; Deme, S


    Al2O3:Mg,Y ceramic thermoluminescence dosemeters were developed at the Institute of Isotopes for high dose applications at room temperatures. The glow curve of Al2O3:Mg,Y exhibits two peaks--one at 250 degrees C (I) and another peak at approximately 400 degrees C (II). In order to extend the application of these dosemeters to high temperatures, the effect of irradiation temperature was investigated using temperature controlled heating system during high dose irradiation at various temperatures (20-100 degrees C). The new calibration and measuring method has been successfully applied for dose mapping within the hermetic zone of the Paks Nuclear Power Plant even at high temperature parts of blocks.

  3. Whey protein hydrolysate enhances HSP90 but does not alter HSP60 and HSP25 in skeletal muscle of rats.

    Directory of Open Access Journals (Sweden)

    Carolina Soares Moura

    Full Text Available Whey protein hydrolysate (WPH intake has shown to increase HSP70 expression. The aim of the present study was to investigate whether WPH intake would also influences HSP90, HSP60 and HSP25 expression, as well as associated parameters. Forty-eight male Wistar rats were divided into sedentary (unstressed and exercised (stressed groups, and were fed with three different sources of protein: whey protein (WP, whey protein hydrolysate (WPH and casein (CAS as a control, based on the AIN93G diet for 3 weeks. WPH intake increased HSP90 expression in both sedentary and exercised animals compared to WP or CAS, however no alteration was found from exercise or diet to HSP60 or HSP25. Co-chaperone Aha1 and p-HSF1 were also increased in the exercised animals fed with WPH in comparison with WP or CAS, consistent with enhanced HSP90 expression. VEGF and p-AKT were increased in the WPH exercised group. No alteration was found in BCKDH, PI3-Kinase (p85, GFAT, OGT or PGC for diet or exercise. The antioxidant system GPx, catalase and SOD showed different responses to diet and exercise. The data indicate that WPH intake enhanced factors related to cell survival, such as HSP90 and VEGF, but does not alter HSP60 or HSP25 in rat skeletal muscle.

  4. ENEA extremity dosemeter based on LiF(Mg,Cu,P) to evaluate Hp(3,alpha). (United States)

    Mariotti, F; Fantuzzi, E; Morelli, B; Gualdrini, G; Botta, M C; Uleri, G; Bordy, J M; Denoziere, M


    Recent epidemiological studies suggest a rather low-dose threshold (ORAMED) Project, whose WP2 is aimed at the quantity H(p)(3) and eye lens dosimetry in practice, are taken into account. The paper summarises the main aspects of the study carried out at ENEA-Radiation Protection Institute (Bologna, Italy) to provide practical solutions (in the use and the design) to evaluate the response of the ENEA TL extremity dosemeter in terms of H(p)(3).


    Silva, E H; Knežević, Ž; Struelens, L; Covens, P; Ueno, S; Vanhavere, F; Buls, N


    Recent studies demonstrated that lens opacities can occur at lower radiation doses than previously accepted. In view of these studies, the International Commission of Radiological Protection recommended in 2011 to reduce the eye lens dose limit from 150 mSv/y to 20 mSv/y. This implies in the need of monitoring doses received by the eye lenses. In this study, small rod radiophotoluminescent glass dosemeters (GD-300 series; AGC, Japan) were characterized in terms of their energy (ISO 4037 X-rays narrow spectrum series, S-Cs and S-Co) and angular dependence (0  up to 90 degrees, with 2 ISO energies: N-60 and S-Cs). All acquisitions were performed at SCK•CEN-Belgium, using the ORAMED proposed cylindrical phantom. For selected energies (N-60, N-80, N-100, N-120 and N-250), the response of dosemeters irradiated on the ISO water slab phantom, at the Ruđer Bošković Institute-Croatia, was compared to those irradiated on the cylindrical phantom. GD-300 series showed good energy dependence, relative to S-Cs, on the cylindrical phantom. From 0 up to 45 degrees, the dosemeters showed no significant angular dependence, regardless whether they were tested when placed vertically or horizontally on the cylindrical phantom. However, at higher angles, some angular dependence was observed, mainly when the dosemeters were irradiated with low-energy photons (N-60). Results showed that GD-300 series have good properties related to Hp(3), although some improvements may be necessary.

  6. Individual monitoring in nuclear medicine therapeutic procedures using extremity dosemeters LiF(Mg, Cu, P). (United States)

    Sarti, G; Del Dottore, F; Fabbri, C; Tassinari, L; Pagan, S; Rustignoli, M; Motta, P


    Unsealed beta-gamma-emitting sources are used (15 GBq (90)Y each session) in nuclear medicine therapeutic procedures. Inside the manipulation cell and while giving the injection to the patient, the skin exposure is very high; electron radiation field is not homogeneous and thus the exposure of the extremities is not uniform. Particular individual monitoring is adopted: single thermoluminescence dosemeter, wrapped in polyethylene film and placed on an adhesive tape, is positioned on the tip of the fingers; 6-10 dosemeters are assigned to each operator per session. The energy and angle response is studied for X-ray spectra, (90)Sr/Y and (204)Tl--a unique mean calibration factor is calculated in order to estimate H(p)(0.07). Performance of dosemeter is analysed according to ISO 62387-1(2007) and the combined uncertainty (calculated using the Monte Carlo method) results lie in the order of 11 %. This method reveals the critical step of manipulation and administration and ensures that dose limits are not exceeded.

  7. The 12B counter: an active dosemeter for high-energy neutrons. (United States)

    Leuschner, A


    High-energy accelerators can produce strong time-structured radiation fields. Such dose shots are generated at linear machines with low duty cycles as well as at circular machines when complete fills are instantaneously lost. The main dose component behind thick shielding is due to high-energy neutrons occurring at that time structure. Dosemeters based on Geiger-Mueller tubes or proportional counters fail here completely. The 12B counter, a novel dosemeter made of a plastic scintillator using carbon activation for event-like exposure, has been introduced. High-energy neutrons activate the carbon nuclei by three inelastic reactions. The decay patterns with half-lives between 20 ms and 20 min can be exploited depending on the time structure of the radiation field. The response of the 12B counter was measured along with some other dosemeters, both active and passive, in the radiation field behind the lateral concrete shielding of a 7.5 GeV proton transfer line.

  8. Hsp10, Hsp70, and Hsp90 immunohistochemical levels change in ulcerative colitis after therapy

    Directory of Open Access Journals (Sweden)

    G. Tomasello


    Full Text Available Ulcerative colitis (UC is a form of inflammatory bowel disease (IBD characterized by damage of large bowel mucosa and frequent extra-intestinal autoimmune comorbidities. The role played in IBD pathogenesis by molecular chaperones known to interact with components of the immune system involved in inflammation is unclear. We previously demonstrated that mucosal Hsp60 decreases in UC patients treated with conventional therapies (mesalazine, probiotics, suggesting that this chaperonin could be a reliable biomarker useful for monitoring response to treatment, and that it might play a role in pathogenesis. In the present work we investigated three other heat shock protein/molecular chaperones: Hsp10, Hsp70, and Hsp90. We found that the levels of these proteins are increased in UC patients at the time of diagnosis and decrease after therapy, supporting the notion that these proteins deserve attention in the study of the mechanisms that promote the development and maintenance of IBD, and as biomarkers of this disease (e.g., to monitor response to treatment at the histological level.

  9. Hsp90 inhibition accelerates cell lysis. Anti-Hsp90 ribozyme reveals a complex mechanism of Hsp90 inhibitors involving both superoxide- and Hsp90-dependent events. (United States)

    Sreedhar, Amere Subbarao; Mihály, Katalin; Pató, Bálint; Schnaider, Tamás; Steták, Attila; Kis-Petik, Katalin; Fidy, Judit; Simonics, Tibor; Maraz, Anna; Csermely, Péter


    The 90 kDa heat shock protein, Hsp90, is an abundant molecular chaperone participating in the cytoprotection of eukaryotic cells. Here we analyzed the involvement of Hsp90 in the maintenance of cellular integrity using partial cell lysis as a measure. Inhibition of Hsp90 by geldanamycin, radicicol, cisplatin, and novobiocin induced a significant acceleration of detergent- and hypotonic shock-induced cell lysis. The concentration and time dependence of cell lysis acceleration was in agreement with the Hsp90 inhibition characteristics of the N-terminal inhibitors, geldanamycin and radicicol. Glutathione and other reducing agents partially blocked geldanamycin-induced acceleration of cell lysis but were largely ineffective with other inhibitors. Indeed, geldanamycin treatment led to superoxide production and a change in membrane fluidity. When Hsp90 content was diminished using anti-Hsp90 hammerhead ribozymes, an accelerated cell lysis was also observed. Hsp90 inhibition-induced cell lysis was more pronounced in eukaryotic (yeast, mouse red blood, and human T-lymphoma) cells than in bacteria. Our results indicate that besides the geldanamycin-induced superoxide production, and a consequent increase in cell lysis, inhibition or lack of Hsp90 alone can also compromise cellular integrity. Moreover, cell lysis after hypoxia and complement attack was also enhanced by any type of Hsp90 inhibition used, which shows that the maintenance of cellular integrity by Hsp90 is important in physiologically relevant lytic conditions of tumor cells.

  10. Effective dose assessment in the maxillofacial region using thermoluminescent (TLD) and metal oxide semiconductor field-effect transistor (MOSFET) dosemeters: a comparative study

    NARCIS (Netherlands)

    Koivisto, J.; Schulze, D.; Wolff, J.; Rottke, D.


    Objectives: The objective of this study was to compare the performance of metal oxide semiconductor field-effect transistor (MOSFET) technology dosemeters with thermoluminescent dosemeters (TLDs) (TLD 100; Thermo Fisher Scientific, Waltham, MA) in the maxillofacial area. Methods: Organ and effective


    Directory of Open Access Journals (Sweden)

    E. N. Sheleenkova


    Full Text Available The article gives a comparative analysis of existing and newly developed thermoluminescent detectors DTG-4, TTLD-580, TLD-1011(TM and DTVS-01 and dosemeters MKD-A and MKD-B for the purposes of individual dosimetric control of skin and eye lens exposure to radiation. The dependence of sensitivity of different thermoluminescent dosemeters, designed for dose equivalents measurements in skin and eye lens on the type (photons and ȕ-particles and energy of radiation was determined experimentally. There are conclusions about the limits of applicability of existing detectors and dosemeters under different radiation exposure conditions. The article evaluates the possibility of different dosimeter use for specific radiation exposure conditions.

  12. Response of neutron dosemeters in radiation protection environments: an investigation of techniques to improve estimates of dose equivalent

    Energy Technology Data Exchange (ETDEWEB)

    Naismith, O.F.; Thomas, D.J. [National Physical Lab., Teddington (United Kingdom); Siebert, B.R.L. [Physikalisch-Technische Bundesanstalt, Braunschweig (Germany)


    The response of practicable neutron dosemeters for routine use generally does not match the conversion function from fluence for radiation protection quantities such as the ambient dose equivalent. As a consequence, significant errors may be encountered when monitoring in a neutron energy spectrum different from that in which the dosemeter was calibrated, which is almost inevitably the case. A database of neutron energy spectra, detector response functions, and dosimetric conversion factors has been developed, and has been used to investigate the extent of this problem. The paper examines various ways of improving dosemeter response by `ranking` spectra and deriving correction factors based upon this ordering. In the case of area monitoring, a combination of two responses (e.g. a rem meter and TEPC) may serve to improve the measurement of dose equivalent. (author).

  13. Characterisation of the Plasmodium falciparum Hsp70-Hsp90 organising protein (PfHop). (United States)

    Gitau, Grace W; Mandal, Pradipta; Blatch, Gregory L; Przyborski, Jude; Shonhai, Addmore


    Malaria is caused by Plasmodium species, whose transmission to vertebrate hosts is facilitated by mosquito vectors. The transition from the cold blooded mosquito vector to the host represents physiological stress to the parasite, and additionally malaria blood stage infection is characterised by intense fever periods. In recent years, it has become clear that heat shock proteins play an essential role during the parasite's life cycle. Plasmodium falciparum expresses two prominent heat shock proteins: heat shock protein 70 (PfHsp70) and heat shock protein 90 (PfHsp90). Both of these proteins have been implicated in the development and pathogenesis of malaria. In eukaryotes, Hsp70 and Hsp90 proteins are functionally linked by an essential adaptor protein known as the Hsp70-Hsp90 organising protein (Hop). In this study, recombinant P. falciparum Hop (PfHop) was heterologously produced in E. coli and purified by nickel affinity chromatography. Using specific anti-PfHop antisera, the expression and localisation of PfHop in P. falciparum was investigated. PfHop was shown to co-localise with PfHsp70 and PfHsp90 in parasites at the trophozoite stage. Gel filtration and co-immunoprecipitation experiments suggested that PfHop was present in a complex together with PfHsp70 and PfHsp90. The association of PfHop with both PfHsp70 and PfHsp90 suggests that this protein may mediate the functional interaction between the two chaperones.

  14. Time-Dependent and Light-Induced Fading in Victoreen Model 2600-80 Aluminum Oxide Thermoluminescence Dosemeters (United States)


    2600-80 Aluminium Oxide Thermoluminescence Dosemeters ":"TIC,7 • :+ " 7-t-•CTE S , E 1P 1993 F’ t LEA l E J. H. Musk AFRRI TR93-6 Cleared for Public...SUBTITLE 5. FUNDING NUMBERS Time-Dependent and Light-Induced Fading in Victoreen0 Model 2600-80 Aluminium Oxide Thermoluminescence PE: QAXM...MODEL 2600-80 ALUMINIUM OXIDE THERMOLUMINESCENCE DOSEMETERS J. H. Musk Accesion For NTIS CRA&I DTIC TAB U.,announced D Justification ........ -.. By

  15. Optical fibre dosemeter systems for clinical applications based on radioluminescence and optically stimulated luminescence from Al2O3:C

    DEFF Research Database (Denmark)

    Marckmann, C.J.; Andersen, C.E.; Aznar, M.C.


    Optical fibre dosemeter systems based on radioluminescence and optically stimulated luminescence (OSL) from carbon-doped aluminium oxide (Al2O3:C) crystals were developed for in vivo real-time dose rate and absorbed dose measurements in radiotherapy and mammography. A technique was also developed...... for making ultra-small dosemeter probes that can easily be placed inside patients in radiation treatment. These probes have shown excellent properties in both head and neck intensity-modulated radiation therapy treatment and in mammography. The dose-response of the OSL signal for the new optical fibre...

  16. Plasmodium falciparum Hsp70-z, an Hsp110 homologue, exhibits independent chaperone activity and interacts with Hsp70-1 in a nucleotide-dependent fashion. (United States)

    Zininga, Tawanda; Achilonu, Ikechukwu; Hoppe, Heinrich; Prinsloo, Earl; Dirr, Heini W; Shonhai, Addmore


    The role of molecular chaperones, among them heat shock proteins (Hsps), in the development of malaria parasites has been well documented. Hsp70s are molecular chaperones that facilitate protein folding. Hsp70 proteins are composed of an N-terminal nucleotide binding domain (NBD), which confers them with ATPase activity and a C-terminal substrate binding domain (SBD). In the ADP-bound state, Hsp70 possesses high affinity for substrate and releases the folded substrate when it is bound to ATP. The two domains are connected by a conserved linker segment. Hsp110 proteins possess an extended lid segment, a feature that distinguishes them from canonical Hsp70s. Plasmodium falciparum Hsp70-z (PfHsp70-z) is a member of the Hsp110 family of Hsp70-like proteins. PfHsp70-z is essential for survival of malaria parasites and is thought to play an important role as a molecular chaperone and nucleotide exchange factor of its cytosolic canonical Hsp70 counterpart, PfHsp70-1. Unlike PfHsp70-1 whose functions are fairly well established, the structure-function features of PfHsp70-z remain to be fully elucidated. In the current study, we established that PfHsp70-z possesses independent chaperone activity. In fact, PfHsp70-z appears to be marginally more effective in suppressing protein aggregation than its cytosol-localized partner, PfHsp70-1. Furthermore, based on coimmunoaffinity chromatography and surface plasmon resonance analyses, PfHsp70-z associated with PfHsp70-1 in a nucleotide-dependent fashion. Our findings suggest that besides serving as a molecular chaperone, PfHsp70-z could facilitate the nucleotide exchange function of PfHsp70-1. These dual functions explain why it is essential for parasite survival.

  17. Expression of heat shock proteins (HSP27, HSP60, HSP70, HSP90,GRP78, GRP94) in hepatitis B virus-related hepatocellular carcinomas and dysplastic nodules

    Institute of Scientific and Technical Information of China (English)

    Seung Oe Lim; Cheol Keun Park; Sung Gyoo Park; Jun-Hi Yoo; Young Min Park; Hie-Joon Kim; Kee-Taek Jang; Jae Won Cho; Byung Chul Yoo; Gu-Hung Jung


    AIM: Expression of heat shock proteins (HSPs) is frequently up-regulated in hepatocellular carcinoma (HCC), which evolves from dysplastic nodule (DN) and early HCC to advanced HCC. However, little is known about the differential expression of HSPs in multistep hepatocarcinogenesis. It was the purpose of this study to monitor the expression of HSPs in multistep hepatocarcinogenesis and to evaluate their prognostic significance in hepatitis B virus (HBV)related HCC.METHODS: Thirty-eight HCC and 19 DN samples were obtained from 52 hepatitis B surface antigen-positive Korean patients. Immunohistochemical and dot immunoblot analyses of HSP27, HSP60, HSP70, HSP90, glucoseregulated protein (GRP)78, and GRP94 were performed and their expression at different stages of HCC development was statistically analyzed.RESULTS: Expression of HSP27, HSP70, HSP90, GRP78, and GRP94 increased along with the stepwise progression of hepatocarcinogenesis. Strong correlation was found only in GRP78 (Spearman's r= 0.802). There was a positive correlation between the expressions of GRP78, GRP94, HSP90, or HSP70 and prognostic factors of HCC. Specifically, the expression of GRP78, GRP94, or HSP90 was associated significantly with vascular invasion and intrahepatic metastasis.CONCLUSION: The expressions of HSPs are commonly up-regulated in HBV-related HCCs and GRP78 might play an important role in the stepwise progression of HBVrelated hepatocarcinogenesis. GRP78, GRP94, and HSP90 may be important prognostic markers of HBV-related HCC, strongly suggesting vascular invasion and intrahepatic metastasis.

  18. Expression of stress response HSP70 gene in Asian paddle crabs, Charybdis japonica, exposure to endocrine disrupting chemicals, bisphenol A (BPA) and 4-nonylphenol (NP) (United States)

    Park, Kiyun; Kwak, Ihn-Sil


    The Asian paddle crab, Charybdis japonica, is a potential bio-indicator reflecting marine sediment toxicity as well as a commercially important species living along coastal areas in Korea. This study investigated its stress response by looking at the heat shock protein (HSP70) gene of C. japonica when the organism is exposed to bisphenol A (BPA) and 4-nonylphenol (NP). We characterized partial sequence of HSP70 as the stressresponse gene of C. japonica. The nucleotide sequence of C. japonica HSP70 is over 90% homologous with the corresponding gene of other crabs. Phylogenetic tree analysis revealed a close relationship between C. japonica HSP70 and HSP70 in other species of lobster and shrimps. HSP70 mRNA transcripts were detected in all the examined tissues of C. japonica, with the highest level in gills, the organ that most frequently came into contact with the external BPA or NP-laden water. As no reference data were available for C. japonica crab exposure, the BPA and NP 24-h LC50 values have not been previously determined. The expression of the C. japonica HSP70 gene to various BPA or NP concentrations during short and longer times was assessed. Gene expression was significantly induced in concentration- and time-dependent manners after BPA or NP exposures. These results support the postulation that crab C. japonica HSP70 could be a potential stress response molecular marker to monitor marine ecosystems.

  19. Pleurotus sajor-caju HSP100 complements a thermotolerance defect in hsp104 mutant Saccharomyces cerevisiae

    Indian Academy of Sciences (India)

    Jin-Ohk Lee; Mi-Jeong Jeong; Tack-Ryun Kwon; Seung-Kon Lee; Myung-Ok Byun; Ill-Min Chung; Soo-Chul Park


    A putative Hsp100 gene was cloned from the fungus Pleurotus sajor-caju. mRNA expression studies demonstrated that this gene (designated PsHsp100) is highly induced by high temperature, induced less strongly by exposure to ethanol, and not induced by drought or salinity. Heat shock induction is detectable at 37°C and reaches a maximum level at 42°C. PsHsp100 mRNAlevels sharply increased within 15 min of exposure to high temperature, and reached a maximum expression level at 2 h that was maintained for several hours. These results indicate that PsHsp100 could work at an early step in thermotolerance. To examine its function, PsHsp100 was transformed into a temperature-sensitive hsp104 deletion mutant Saccharomyces cerevisiae strain to test the hypothesis that PsHSP100 is an protein that functions in thermotolerance. Overexpression of PsHSP100 complemented the thermotolerance defect of the hsp104 mutant yeast, allowing them survive even at 50°C for 4 h. These results indicate that PsHSP100 protein is functional as an HSP100 in yeast and could play an important role in thermotolerance in P. sajor-caju.

  20. Hsp25 and Hsp72 content in rat skeletal muscle following controlled shortening and lengthening contractions. (United States)

    Holwerda, Andrew M; Locke, Marius


    The cytoprotective proteins, Hsp25 and Hsp72, are increased in skeletal muscle after nondamaging, shortening contractions, but the temporal pattern of expression and stimulatory mechanisms remain unclear. Thus, we sought to define the in vivo temporal patterns of expression for Hsp25 and Hsp72 after 2 opposing contractions types. To do this, male Sprague-Dawley rats had 1 tibialis anterior (TA) muscle electrically stimulated (5 sets of 20 repetitions) while being either forcibly lengthened (LC) or shortened (SC). At 2, 8, 24, 48, 72, or 168 h after the contractions both the stimulated and the nonstimulated (contra-lateral control) TA muscles were removed and processed to examine muscle damage (hemotoxylin and eosin staining) and Hsp content (Western blot analyses). Cross-sections from TA muscles subjected to LCs showed muscle fibre damage at 8 h and thereafter. In contrast, no muscle fibre damage was observed at any time point following SCs. When normalized to contra-lateral controls, Hsp25 and Hsp72 content were significantly (P < 0.01) increased at 24 h (3.1- and 3.8-fold, respectively) and thereafter. There were no significant increases in Hsp25 or Hsp72 content at any time point following SC. These data suggest that LCs, but not SCs, result in Hsp accumulation and that the fibre/cellular damage sustained from LCs may be the stimulus for elevating Hsp content.

  1. Pleurotus sajor-caju HSP100 complements a thermotolerance defect in hsp104 mutant Saccharomyces cerevisiae

    Indian Academy of Sciences (India)

    Jin-Ohk Lee; Mi-Jeong Jeong; Tack-Ryun Kwon; Seung-Kon Lee; Myung-Ok Byun; Ill-Min Chung; Soo-Chul Park


    A putative Hsp100 gene was cloned from the fungus Pleurotus sajor-caju. mRNA expression studies demonstrated that this gene (designated PsHsp100) is highly induced by high temperature, induced less strongly by exposure to ethanol, and not induced by drought or salinity. Heat shock induction is detectable at 37°C and reaches a maximum level at 42°C. PsHsp100 mRNAlevels sharply increased within 15 min of exposure to high temperature, and reached a maximum expression level at 2 h that was maintained for several hours. These results indicate that PsHsp100 could work at an early step in thermotolerance. To examine its function, PsHsp100 was transformed into a temperature-sensitive hsp104 deletion mutant Saccharomyces cerevisiae strain to test the hypothesis that PsHSP100 is an protein that functions in thermotolerance. Overexpression of PsHSP100 complemented the thermotolerance defect of the hsp104 mutant yeast, allowing them survive even at 50°C for 4 h. These results indicate that PsHSP100 protein is functional as an HSP100 in yeast and could play an important role in thermotolerance in P. sajor-caju.

  2. Development of advanced-type multi-functional electronic personal dosemeter. (United States)

    Nunomiya, T; Abe, S; Aoyama, K; Nakamura, T


    An advanced-type small, light, multi-functional electronic personal dosemeter has been developed using silicon semiconductor radiation detectors for dose management of workers at nuclear power plants and accelerator facilities. This dosemeter is 62 x 82 x 27 mm(3) in size and approximately 130 g in weight, which is capable of measuring personal gamma ray and neutron dose equivalents, Hp(10), simultaneously. The neutron dose equivalent can be obtained using two types of silicon semiconductors: a slow-neutron sensor (1 MeV). The slow neutron sensor is a 10 x 10 mm(2) p-type silicon on which a natural boron layer is deposited around an aluminium electrode. The fast neutron sensor is also a 10 x 10 mm(2) p-type silicon crystal on which an amorphous silicon hydride is deposited. The neutron energy response corresponding to the fluence-to-dose-equivalent conversion coefficient given by ICRP Publication 74 has been evaluated using a monoenergetic neutron source from 250 keV to 15 MeV at the Fast Neutron Laboratory of Tohoku University. As the result, the Hp(10) response to neutrons in the energy range of 250 keV and 4.4 MeV within +/-50% difference has been obtained.

  3. In-vivo dosimetry in radiotherapy: a comparison of the response of semiconductor and thermoluminescence (TLD700) dosemeters

    Energy Technology Data Exchange (ETDEWEB)

    Vynckier, S. [Universite Catholique de Louvain, Brussels (Belgium). Cliniques Universitaires St. Luc; Greffe, J.L.; Loncol, T.; Vanneste, F.; Octave-Prignot, M.; Denis, J.M.; De Patoul, N.


    Semiconductor dosemeters and thermoluminescence dosemeters were calibrated in view of in-vivo dosimetry. Their response in a 8 MV photon beam and the respective correction factors for the treatment conditions were systematically studied. A total of 249 entrance and exit measurements with this dual detector combination were performed, mainly for treatments of the head and neck region. The resulting entrance and exit doses were compared with the expected doses at these positions, calculated on basis of the treatment and patient parameters. The results at the entrance showed a value of 1.010 (2.8% for the ratio of the measured to the calculated dose by diodes, 1.013) 4.9% for the ratio of the measured to the calculated dose by TLD and 1.003 (3.6% for the ratio of the measured dose by TLD to diodes. With respect the exit dose, the results were 0.998) 4.9%, 1.016 (7.7% and 1.019) 7.0% respectively after correction for the heterogeneity`s. Although the standard deviation for the TLD dosemeters is systematically larger than the standard deviation for the diodes, it is concluded that both dosemeters will yield similar results for-in-vivo dosimetry, if utilized under the same conditions.

  4. Viability of the Fricke dosemeter doped with methylene blue; Viabilidade do dosimetro Fricke dopado com azul de metileno

    Energy Technology Data Exchange (ETDEWEB)

    Souza, V.L.B.; Santos, C.D.A.; Rodrigues, K.R.G.; Cunha, M.S.; Figueiredo, M.D.C.; Melo, R.T. [Centro Regional de Ciencias Nucleares do Nordeste (CRCN-NE/CNEN-PE), Recife, PE (Brazil)


    This work aims to find the possible utilization of the Fricke dosemeter doped with methylene blue (FMB) for the dosimetry of photodynamic therapy. The FMB was irradiated wit X rays and light emitted diodes demonstrating positive answers to the stimulus, being probably to be used for dosimetric objectives


    Rizk, C; Vanhavere, F


    The personal dosimetry service at the Lebanese Atomic Energy Commission uses Harshaw 8814 cards with LiF:Mg,Ti detectors. The dosemeters are read in a Harshaw 6600 TLD reader. In the process of accreditation for the ISO 17025 standard((1)), different influence factors are investigated and the uncertainty has been determined. The Individual Monitoring Service Laboratory-LAEC reads the dosemeters once it receives them from the customer, and new cards are immediately given for the next wearing period. The wearing period is 2 months. The dosemeter results are reported to the customers without background subtraction. Both Hp(10) and Hp(0.07) are reported. For this paper, only the uncertainty on Hp(10) will be focussed. The following factors are taken into account for the uncertainty: calibration factor, dosemeter homogeneity and repeatability, energy and angular dependence, non-linearity, temperature dependence, etc. Also the detection limit was determined. One of the important factors is the correction for fading. This fading correction depends on the procedure used such as storage temperatures, the time-temperature profile of the read-out, pre-heat and annealing conditions. Pre- and post-irradiation fading curves were measured for a storage period up to 182 d at room temperature (15-25°C). The resulting final combined standard uncertainty on the reported doses is of the order of 24 % for doses of ∼1 mSv.

  6. MCNP simulation of the incident and Albedo neutron response of the IRD Albedo Neutron Dosemeter for {sup 241}Am-Be moderated sources

    Energy Technology Data Exchange (ETDEWEB)

    Freitas, Bruno M.; Martins, Marcelo M.; Mauricio, Claudia L.P.; Mauricio, Claudia L.P. da, E-mail: [Instituto de Radioprotecao e Dosimetria (IRD/CNEN-RJ), Rio de Janeiro, RJ (Brazil); Silva, Ademir X. [Coordenacao dos Programas de Pos-Graduacao em Engenharia (COPPE/UFRJ), Rio de Janeiro, RJ (Brazil). Programa de Engenharia Nuclear


    The IRD TLD Albedo dosemeter measures both incident and albedo neutron component. The incident to Albedo ratio is used to take into account the energy dependence of its response. In this paper, the behavior of the IRD Albedo dosemeter response as a function of the incident to Albedo ratio for {sup 241}Am-Be sources was simulated to improve its algorithm. The simulation was performed in MCNPX transport code and presents a good agreement with experimental measurements. The results obtained in this work are very useful to improve the accuracy of the IRD Albedo dosemeter at real neutron workplace. (author)

  7. Hsp70 Structure, Function, Regulation and Influence on Yeast Prions


    D. Sharma; Masison, D. C.


    Heat shock proteins protect cells from various conditions of stress. Hsp70, the most ubiquitous and highly conserved Hsp, helps proteins adopt native conformation or regain function after misfolding. Various co-chaperones specify Hsp70 function and broaden its substrate range. We discuss Hsp70 structure and function, regulation by co-factors and influence on propagation of yeast prions.

  8. Echinococcus granulosus: in vitro effects of ivermectin and praziquantel on hsp60 and hsp70 levels. (United States)

    Martinez, J; Perez-Serrano, J; Bernadina, W E; Rodriguez-Caabeiro, F


    Martinez, J., Perez-Serrano, J., Bernadina, W. E., Rodriguez-Caabeiro, F. 1999 Echinococcus granulosus: In vitro effects of ivermectin and praziquantel on hsp60 and hsp70 levels. Experimental Parasitology93, 171-180. Organisms or cells exposed to injurious stresses such as heat shock or chemicals respond by increased (or altered) expression of heat-shock proteins (HSPs). Conversely, an earlier exposure to stress can prepare cells to cope with a subsequent more severe stress. In the present study, protoscolices of Echinococcus granulosus were subjected to several anthelmintic treatments, involving storage of the protoscolices for 18, 30, and 50 h with 0.1 mg/ml of ivermectin (IV), praziquantel (PZ), and a combination of each with albendazole (ALB). The organisms were analyzed for the effects of drug treatment on cell integrity and on levels of hsp60 and hsp70 production. Drug efficacy was evaluated by microscopy and by protein content measurement. Hsp60 and hsp70 were detected by Western blotting and incubation with anti-hsp60 and anti-hsp70 antibody, respectively, and quantitation of these proteins was obtained using image analysis. Incubation with IV alone produced the most damage to the protoscolices as indicated by viability loss, decreased protein content, and altered hsp60 and hsp70 levels; incubation with IV + ALB produced less damage as manifested by fewer changes in the aforementioned damage parameters but PZ and PZ + ALB, in this context, were poor anthelmintics. Exposure of protoscolices to thermal stress prior to anthelmintic treatment, in most cases, increased drug efficacy. It is concluded that in the E. granulosus model system drug efficacy is associated with decreased levels of hsp70 expression and increased levels of hsp60 expression.

  9. Performance of Panasonic ZP-1460 Electronic Personal Dosemeter under Exposure Conditions Likely to be Found at Fukushima Daiichi Nuclear Power Plant (United States)

    Tsujimura, Norio; Yoshida, Tadayoshi; Hoshi, Katsuya; Momose, Takumaro

    A study on the performance of the Panasonic ZP-1460 electronic personal dosemeter, the model used in the aftermath of the Fukushima Daiichi nuclear power plant accident in March 2011, was conducted under actual exposure situations likely encountered in the plant. The tests pertained to (1) the dose rate response over dose rates >100 mSv/h and (2) the angular response on an anthropomorphic phantom exposed to the rotational and isotropic irradiation geometries. The test results confirmed that the dosemeter provides Hp(10) as a reasonably close estimate of the effective dose for any exposure geometries. The dosemeter response data evaluated in this study can be utilized for converting dosemeter readings to the absorbed dose to any organs and tissues for epidemiologic purposes.

  10. Ten years of personnel neutron dosimetry with Albedo dosemeters in the Netherlands

    Energy Technology Data Exchange (ETDEWEB)

    Draaisma, F.S.; Verhagen, H.W


    Since 1987, the dosimetry service of the Netherlands Energy Research Foundation has been certified by the Dutch government to perform personal dosimetry, using thermoluminescence dosemeters (TLDs). Performing neutron personal dosimetry requires a rather large investment in readers, TLDs and personnel to operate the service. About 800 persons are subjected to routine neutron monitoring in the Netherlands and their annual neutron doses are a relatively small fraction (less than 10%) of the annual H{sub p}(10). In general, the measured neutron dose values are low (on average 93% of the users receive an annual neutron dose <0.2 mSv). The collective annual (neutron) dose has tended to decrease since 1992, but incidentally high doses have been observed. Leaving these incidents out, the average collective annual neutron doses for the different users of neutron sources are about the same. (author)

  11. Performance of the EPD-N2 dosemeter for monitoring aircrew doses. (United States)

    Scherpelz, R I; Cezeaux, J R


    United States Air Force (USAF) aircrew fly at altitudes and for durations where doses from cosmic radiation are significant enough to warrant monitoring. This study evaluated a candidate radiological monitoring system for USAF aircrew, the Thermo Scientific electronic personnel dosemeter (EPD-N2). The evaluation consisted of characterising the device in a well-characterised radiation field at a European Organization for Nuclear Research (CERN) accelerator, and aboard an USAF aircraft. The performance of the EPDs was evaluated by comparison with accepted values for dose at the CERN facility, comparison with the value calculated by flight dose software and comparison with the value estimated by a tissue-equivalent proportional counter aboard the aircraft. This study recommends that a correction factor of 1/CF = 1/3.9 be applied to EPD-N2 measurements aboard aircraft flights. The uncertainty in this correction factor is 11.8 %.

  12. Measurement analysis using the Fricke dosemeter; Analise de medicoes com o dosimetro Fricke

    Energy Technology Data Exchange (ETDEWEB)

    Pedrosa, G.A.C. [Instituto Nacional de Metrologia (INMETRO), Rio de Janeiro, RJ (Brazil)], e-mail:; Dantas, C.C. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Energia Nuclear], e-mail:; Campos, C.A. [Centro Regional de Ciencias Nucleares do Nordeste (CRCN-NE/CNEN-PE), Recife, PE (Brazil)], e-mail:


    The Fricke dosemeter has been used for absolute determination of the radiation dose between 40 and 350 Gy for X-rays, Gamma rays and fast electrons energies. The great inconvenient of that dosimetric system is the impossibility of using for measurements of exposure or low dose rates. A Fricke dosimetry system was implanted at the Nuclear Sciences Regional Center (CRCN), in Recife, Pernambuco state, Brazil, for the determination of radiation doses for spectrometry and mass dosimetry with plasma source. The statistical analysis of the obtained results have shown that using the spectrometric method that system cna be used for maintenance of factors of CRCN standard calibrations and the dose assessment thru mail system of radiotherapy centers of the country.


    Cardoso, J; Santos, J A M; Santos, L; Alves, J G; Oliveira, C


    The active personal dosemeter, RaySafe i2, allows the measurement and record of Hp(10) in real time, every second, via wireless technology for real-time display on a portable computer and/or a local network. The system seems particularly attractive for individual monitoring at clinical facilities where high intensity and varying radiation fields may occur, as it enables the user to acknowledge and optimize the dose and dose rate values in real time for each procedure. Prior to its use, the system was characterized at the Metrology Laboratory of Ionizing Radiation of IST-LPSR aiming at the metrological characterization of the system in accordance with IEC 61526:2010 for metrological control purposes and to verify the technical specifications stated by the manufacturer.

  14. Thermoluminescent dosemeter in a X-ray diffractometer; Dosimetria termoluminiscente en un difractometro de rayos X

    Energy Technology Data Exchange (ETDEWEB)

    Mendoza A, D.; Gonzalez M, P.; Falcon B, T. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, Mexico D.F. (Mexico); Castano, V.M. [Instituto de Fisica, UNAM, Laboratorio Juriquilla, A.P. 1-1010, C.P. 76001, Queretaro (Mexico)


    In this work it was presented the results obtained of the dosimetry which was realized in a X-ray diffractometer for powders, trademark Siemens D5000, using the thermoluminescent signal generated by the X-rays in the commercial dosemeter TLD-100 of Harshaw, US. In according to the results obtained, the radiation quantity received by an analysed material in the diffractometer, will be proportional to exposure time and it can vary from unities until tenths of grays. These results are very outstanding when are analysed crystalline materials in a diffractometer, for knowing the present crystalline phases, mainly if these are highly sensitive to the ionizing radiation, as it is the case of the thermoluminescent materials. (Author)

  15. Heat shock protein genes (hsp20, hsp75 and hsp90) from Pieris rapae: Molecular cloning and transcription in response to parasitization by Pteromalus puparum

    Institute of Scientific and Technical Information of China (English)

    Jia-Ying Zhu; Guo-Xing Wu; Gong-Yin Ye; Cui Hu


    Most molecular work on the roles of heat shock proteins (hsps) in host-parasite interaction has focused on vertebrates,rather than invertebrates.Here the full length complementary DNA (cDNA) sequences of three hsp genes (hsp20,hsp75 and hsp90)were amplified from Pieris rapae,and their transcriptional responsiveness to parasitization by the endoparasitic wasp Pteromalus puparum were investigated.The cDNA sequence analysis of hsp20,hsp75 and hsp90 revealed open reading frames of 531,2 328 and 2 157 bp in length,which encode proteins with calculated molecular weights of 19.5,75.48 and 82.7 kDa,respectively.The comparison of amino acid sequences showed that P.rapae hsp20 shared highly divergent homology to that of other insects,while hsp75 and hsp90 showed high homology to their counterparts of other species.The expression analysis indicated that these three genes were influenced in response to parasitization by P.puparum.The hsp20 transcripts in parasitized pupae were higher compared to nonparasitized pupae.The expression of hsp75 and hsp90 were down-regulated following parasitization.The results indicate that hsps are involved in host-parasitoid interactions.

  16. HSP86 and HSP84 exhibit cellular specificity of expression and co-precipitate with an HSP70 family member in the murine testis (United States)

    Gruppi, C. M.; Wolgemuth, D. J.


    This study extends to the protein level our previous observations, which had established the stage and cellular specificity of expression of hsp86 and hsp84 in the murine testis in the absence of exogenous stress. Immunoblot analysis was used to demonstrate that HSP86 protein was present throughout testicular development and that its levels increased with the appearance of differentiating germ cells. HSP86 was most abundant in the germ cell population and was present at significantly lower levels in the somatic cells. By contrast, the HSP84 protein was detected in the somatic cells of the testis rather than in germ cells. The steady-state levels of HSP86 and HSP84 paralleled the pattern of the expression of their respective mRNAs, suggesting that regulation at the level of translation was not a major mechanism controlling hsp90 gene expression in testicular cells. Immunoprecipitation analysis revealed that a 70-kDa protein coprecipitated with the HSP86/HSP84 proteins in testicular homogenates. This protein was identified as an HSP70 family member by immunoblot analysis, suggesting that HSP70 and HSP90 family members interact in testicular cells.

  17. Abnormal interaction of motor neuropathy-associated mutant HspB8 (Hsp22) forms with the RNA helicase Ddx20 (gemin3)

    NARCIS (Netherlands)

    Sun, Xiankui; Fontaine, Jean-Marc; Hoppe, Adam D.; Carra, Serena; DeGuzman, Cheryl; Martin, Jody L.; Simon, Stephanie; Vicart, Patrick; Welsh, Michael J.; Landry, Jacques; Benndorf, Rainer


    A number of missense mutations in the two related small heat shock proteins HspB8 (Hsp22) and HspB1 (Hsp27) have been associated with the inherited motor neuron diseases (MND) distal hereditary motor neuropathy and Charcot-Marie-Tooth disease. HspB8 and HspB1 interact with each other, suggesting tha

  18. Determination of isodose curves in Radiotherapy using an Alanine/ESR dosemeter; Determinacion de curvas de isodosis en Radioterapia usando un dosimetro de Alanina/ESR

    Energy Technology Data Exchange (ETDEWEB)

    Chen, F.; Baffa, O.; Graeff, C.F.O. [Departamento de Fisica e Matematica. Universidade de Sao Paulo FFCLRP. 14040-901 Ribeirao Preto-SP (Brazil)


    It was studied the possible use of an Alanine/ESR dosemeter in the isodose curves mapping in normal treatments of Radiotherapy. It was manufactured a lot of 150 dosemeters with base in a mixture of D-L Alanine dust (80 %) and paraffin (20 %). Each dosemeter has 4.7 mm diameter and 12 mm length. A group of 100 dosemeters of the lot were arranged inside 50 holes of the slice 25 of the phantom Rando Man. The phantom irradiation was realized in two opposed projections (AP and PA) in Co-60 equipment. A group of 15 dosemeters was take of the same lot for obtaining the calibration curve in a 1-20 Gy range. After irradiation the signal of each dosemeter was measured in an ESR spectrometer operating in the X-band ({approx} 9.5 GHz) and the wideness of Alanine ESR spectra central line was correlated with the radiation dose. The wideness dose calibration curve resulted linear with a correlation coefficient 0.9996. The isodose curves obtained show a profile enough similar at comparing with the theoretical curves. (Author)

  19. Study of a selection of 10 historical types of dosemeter: variation of the response to Hp(10) with photon energy and geometry of exposure. (United States)

    Thierry-Chef, I; Pernicka, F; Marshall, M; Cardis, E; Andreo, P


    An international collaborative study of cancer risk among workers in the nuclear industry is tinder way to estimate direetly the cancer risk following protracted low-dose exposure to ionising radiation. An essential aspect of this study is the characterisation and quantification of errors in available dose estimates. One major source of errors is dosemeter response in workplace exposure conditions. Little information is available on energy and geometry response for most of the 124 different dosemeters used historically in participating facilities. Experiments were therefore set up to assess this. using 10 dosemeter types representative of those used over time. Results show that the largest errors were associated with the response of early dosemeters to low-energy photon radiation. Good response was found with modern dosemeters. even at low energy. These results are being used to estimate errors in the response for each dosemeter type, used in the participating facilities, so that these can be taken into account in the estimates of cancer risk.

  20. Hsp90: Friends, clients and natural foes. (United States)

    Verma, Sharad; Goyal, Sukriti; Jamal, Salma; Singh, Aditi; Grover, Abhinav


    Hsp90, a homodimeric ATPase, is responsible for the correct folding of a number of newly synthesized polypeptides in addition to the correct folding of denatured/misfolded client proteins. It requires several co-chaperones and other partner proteins for chaperone activity. Due to the involvement of Hsp90-dependent client proteins in a variety of oncogenic signaling pathways, Hsp90 inhibition has emerged as one of the leading strategies for anticancer chemotherapeutics. Most of Hsp90 inhibitors blocks the N terminal ATP binding pocket and prevents the conformational changes which are essential for the loading of co-chaperones and client proteins. Several other inhibitors have also been reported which disrupt chaperone cycle in ways other than binding to N terminal ATP binding pocket. The Hsp90 inhibition is associated with heat shock response, mediated by HSF-1, to overcome the loss of Hsp90 and sustain cell survival. This review is an attempt to give an over view of all the important players of chaperone cycle.

  1. Functional analysis of Hsp70 inhibitors.

    Directory of Open Access Journals (Sweden)

    Rainer Schlecht

    Full Text Available The molecular chaperones of the Hsp70 family have been recognized as targets for anti-cancer therapy. Since several paralogs of Hsp70 proteins exist in cytosol, endoplasmic reticulum and mitochondria, we investigated which isoform needs to be down-regulated for reducing viability of cancer cells. For two recently identified small molecule inhibitors, VER-155008 and 2-phenylethynesulfonamide (PES, which are proposed to target different sites in Hsp70s, we analyzed the molecular mode of action in vitro. We found that for significant reduction of viability of cancer cells simultaneous knockdown of heat-inducible Hsp70 (HSPA1 and constitutive Hsc70 (HSPA8 is necessary. The compound VER-155008, which binds to the nucleotide binding site of Hsp70, arrests the nucleotide binding domain (NBD in a half-open conformation and thereby acts as ATP-competitive inhibitor that prevents allosteric control between NBD and substrate binding domain (SBD. Compound PES interacts with the SBD of Hsp70 in an unspecific, detergent-like fashion, under the conditions tested. None of the two inhibitors investigated was isoform-specific.

  2. Suramin inhibits Hsp104 ATPase and disaggregase activity.

    Directory of Open Access Journals (Sweden)

    Mariana P Torrente

    Full Text Available Hsp104 is a hexameric AAA+ protein that utilizes energy from ATP hydrolysis to dissolve disordered protein aggregates as well as amyloid fibers. Interestingly, Hsp104 orthologues are found in all kingdoms of life except animals. Thus, Hsp104 could represent an interesting drug target. Specific inhibition of Hsp104 activity might antagonize non-metazoan parasites that depend on a potent heat shock response, while producing little or no side effects to the host. However, no small molecule inhibitors of Hsp104 are known except guanidinium chloride. Here, we screen over 16,000 small molecules and identify 16 novel inhibitors of Hsp104 ATPase activity. Excluding compounds that inhibited Hsp104 activity by non-specific colloidal effects, we defined Suramin as an inhibitor of Hsp104 ATPase activity. Suramin is a polysulphonated naphthylurea and is used as an antiprotozoal drug for African Trypanosomiasis. Suramin also interfered with Hsp104 disaggregase, unfoldase, and translocase activities, and the inhibitory effect of Suramin was not rescued by Hsp70 and Hsp40. Suramin does not disrupt Hsp104 hexamers and does not effectively inhibit ClpB, the E. coli homolog of Hsp104, establishing yet another key difference between Hsp104 and ClpB behavior. Intriguingly, a potentiated Hsp104 variant, Hsp104A503V, is more sensitive to Suramin than wild-type Hsp104. By contrast, Hsp104 variants bearing inactivating sensor-1 mutations in nucleotide-binding domain (NBD 1 or 2 are more resistant to Suramin. Thus, Suramin depends upon ATPase events at both NBDs to exert its maximal effect. Suramin could develop into an important mechanistic probe to study Hsp104 structure and function.

  3. Muscle cell atrophy induced by HSP gene silencing was counteracted by HSP overexpression (United States)

    Choi, Inho; Lee, Joo-Hee; Nikawa, Takeshi; Gwag, Taesik; Park, Kyoungsook; Park, Junsoo

    Heat shock proteins (HSP), as molecular chaperones, are known to assist protein quality control under various stresses. Although overexpression of HSP70 was found to contribute to muscle size retention under an unloading condition, it remains largely unclarified whether muscle atrophy is induced by active suppression of HSP expression. In this study, we pre-treated Hsp70 siRNA to rat L6 cells for the HSP gene silencing, and determined myotube diameter, HSP72 expression and anabolic and catabolic signaling activities in the absence or presence of triterpene celastrol (CEL), the HSP70 inducer. Relative to a negative control (NC), muscle cell diameter was reduced 0.89-fold in the siRNA-treated group, increased 1.2-fold in the CEL-treated group and retained at the size of NC in the siRNA+CEL group. HSP72 expression was decreased 0.35-fold by siRNA whereas the level was increased 6- to 8-fold in the CEL and siRNA+CEL groups. Expression of FoxO3 and atrogin-1 was increased 1.8- to 4.8-fold by siRNA, which was abolished by CEL treatment. Finally, phosphorylation of Akt1, S6K and ERK1/2 was not affected by siRNA, but was elevated 2- to 6-fold in the CEL and siRNA+CEL groups. Taken together, HSP downregulation by Hsp gene silencing led to muscle cell atrophy principally via increases in catabolic activities and that such anti-atrophic effect was counteracted by HSP overexpression.

  4. The nucleotide exchange factors of Hsp70 molecular chaperone

    Directory of Open Access Journals (Sweden)

    Andreas eBracher


    Full Text Available Molecular chaperones of the Hsp70 family form an important hub in the cellular protein folding networks in bacteria and eukaryotes, connecting translation with the downstream machineries of protein folding and degradation. The Hsp70 folding cycle is driven by two types of cochaperones: J-domain proteins stimulate ATP hydrolysis by Hsp70, while nucleotide exchange factors (NEFs promote replacement of Hsp70-bound ADP with ATP. Bacteria and organelles of bacterial origin have only one known NEF type for Hsp70, GrpE. In contrast, a large diversity of Hsp70 NEFs has been discovered in the eukaryotic cell. These NEFs belong to the Hsp110/Grp170, HspBP1/Sil1 and BAG domain protein families. In this short review we compare the structures and molecular mechanisms of nucleotide exchange factors for Hsp70 and discuss how these cochaperones contribute to protein folding and quality control in the cell.

  5. Targeting Hsp90 in Non-Cancerous Maladies. (United States)

    Woodford, Mark R; Dunn, Diana M; Ciciarelli, Joseph G; Beebe, Kristin; Neckers, Len; Mollapour, Mehdi


    Heat shock protein-90 (Hsp90) is a molecular chaperone critical to the folding, stability and activity of over 200 client proteins including many responsible for tumor initiation, progression and metastasis. Hsp90 chaperone function is linked to its ATPase activity and Hsp90 inhibitors interfere with this activity, thereby making Hsp90 an attractive target for cancer therapy. Also post-translational modification (PTM) and co-chaperone proteins modulate Hsp90 function, providing additional targets for secondary inhibition. Recent reports have shown that pathogens utilize both their own Hsp90 and that of their host for the propagation of infectious elements. In this review we will summarize our current knowledge of Hsp90 structure and function in both the pathogen and the host. We will focus on the role of Hsp90 in viral and parasitic diseases and the potential beneficial application of Hsp90 inhibitors alone and in combination with disease-specific inhibitors.

  6. Overexpression, Purification and Characterisation of the Plasmodium falciparum Hsp70-z (PfHsp70-z Protein.

    Directory of Open Access Journals (Sweden)

    Tawanda Zininga

    Full Text Available Six Hsp70-like genes are represented on the genome of Plasmodium falciparum. Of these two occur in the cytosol: P. falciparum Hsp70-z (PfHsp70-z and PfHsp70-1. PfHsp70-1 is a well characterised canonical Hsp70 that facilitates protein quality control and is crucial for the development of malaria parasites. There is very little known about PfHsp70-z. However, PfHsp70-z is known to be essential and is implicated in suppressing aggregation of asparagine-rich proteins of P. falciparum. In addition, its expression at the clinical stage of malaria correlates with disease prognosis. Based on structural evidence PfHsp70-z belongs to the Hsp110 family of proteins. Since Hsp110 proteins have been described as nucleotide exchange factors (NEFs of their canonical Hsp70 counterparts, it has been speculated that PfHsp70-z may serve as a NEF of PfHsp70-1. In the current study, P. falciparum cells cultured in vitro were subjected to heat stress, triggering the enhanced expression of PfHsp70-z. Biochemical assays conducted using recombinant PfHsp70-z protein demonstrated that the protein is heat stable and possesses ATPase activity. Furthermore, we observed that PfHsp70-z is capable of self-association. The structural-functional features of PfHsp70-z provide further evidence for its role as a chaperone and possible nucleotide exchange factor of PfHsp70-1.

  7. Targeting Hsp90-Cdc37: a promising therapeutic strategy by inhibiting Hsp90 chaperone function. (United States)

    Wang, Lei; Li, Li; Gu, Kai; Xu, Xiao-Li; You, Qi-Dong; Sun, Hao-Peng


    The Hsp90 chaperone protein regulates the folding, maturation and stability of a wide variety of oncoproteins. In recent years, many Hsp90 inhibitors have entered into the clinical trials while all of them target ATPase showing similar binding capacity and kinds of side-effects so that none have reached to the market. During the regulation progress, numerous protein-protein interactions (PPI) such as Hsp90 and client proteins or cochaperones are involved. With the Hsp90-cochaperones PPI networks being more and more clear, many cancerous proteins have been reported to be tightly correlated to Hsp90-cochaperones PPI. Among them, Hsp90-Cdc37 PPI has been widely reported to associate with numerous protein kinases, making it a novel target for the treatment of cancers. In this paper, we briefly review the strategies and modulators targeting Hsp90-Cdc37 complex including direct and indirect regulation mechanism. Through these discussions we expect to present inspirations for new insights into an alternative way to inhibit Hsp90 chaperone function.

  8. Comparison of neutron dose quantities and instrument and dosemeter readings at representative locations in an MOX fuel fabrication plant

    CERN Document Server

    Bartlett, D T; Tanner, R J; Haley, R M; Cooper, A J


    The relationships between operational and protection quantities, and values of personal dosemeter and instrument readings have been determined for a recently designed MOX fuel fabrication plant. The relationships between the quantities, and the readings of personal dosemeters are sensitive to both the energy and direction distribution of neutron fluence. The energy distributions were calculated using the Monte Carlo code MCBEND. The direction distribution was addressed by calculating independently, spectral components for which the direction distribution could be reasonably assumed. At representative locations, and for assumed worker orientations, the radiation field is analysed as having, in general, three components--a direct, unidirectional component from the nearest identified discrete source, which is considered incident A-P, several unidirectional components from other such sources which are treated as a rotational component and a scattered isotropic component. The calculated spectra were folded with co...

  9. Measurement of staff doses in interventional procedures using LiF TL-detectors and a special diode dosemeter

    Energy Technology Data Exchange (ETDEWEB)

    Parviainen, T.; Toivonen, M.; Kosunen, A.; Karppinen, J. [Radiation and Nuclear Safety Authority, Helsinki (Finland); Ylitalo, A. [Satakunta Central Hospital, Division for Cardiology, Pori (Finland)


    Personal doses vary considerably among the operational staff in cardiac examination procedures and interventions. Correspondingly, the fluoroscopy time may vary from a few minutes to more than 1 hour. Large personal doses in the range of 35 - 41 mSv were registered to some interventional radiologists and cardiologists in 2001 in Finland. Measurement of doses per intervention from hands, shoulders and ankles (parts of the body outside the lead apron) are therefore of interest, but small and highly sensitive dosemeters have not been commonly available for reliable and practical measurements. Thermoluminescent detectors of LiF:MgCu,P are sensitive enough, but residual luminescence from previous uses of detectors and the dose collected from natural background radiation between preparation for use and readout may be significant sources of uncertainty. The company Unfors Instrumens AB (Billdal, Sweden) has designed a light weight diode dosemeter with long cables between detectors and the electronic part for simultaneous measurement of doses from various parts of the body. The aim of this work is to report doses of the operating staff per a cardiac examination or intervention and study the capabilities of the diode dosemeter in staff dose measurements. (orig.)

  10. Hsp90 inhibition decreases mitochondrial protein turnover.

    Directory of Open Access Journals (Sweden)

    Daciana H Margineantu

    Full Text Available BACKGROUND: Cells treated with hsp90 inhibitors exhibit pleiotropic changes, including an expansion of the mitochondrial compartment, accompanied by mitochondrial fragmentation and condensed mitochondrial morphology, with ultimate compromise of mitochondrial integrity and apoptosis. FINDINGS: We identified several mitochondrial oxidative phosphorylation complex subunits, including several encoded by mtDNA, that are upregulated by hsp90 inhibitors, without corresponding changes in mRNA abundance. Post-transcriptional accumulation of mitochondrial proteins observed with hsp90 inhibitors is also seen in cells treated with proteasome inhibitors. Detailed studies of the OSCP subunit of mitochondrial F1F0-ATPase revealed the presence of mono- and polyubiquitinated OSCP in mitochondrial fractions. We demonstrate that processed OSCP undergoes retrotranslocation to a trypsin-sensitive form associated with the outer mitochondrial membrane. Inhibition of proteasome or hsp90 function results in accumulation of both correctly targeted and retrotranslocated mitochondrial OSCP. CONCLUSIONS: Cytosolic turnover of mitochondrial proteins demonstrates a novel connection between mitochondrial and cytosolic compartments through the ubiquitin-proteasome system. Analogous to defective protein folding in the endoplasmic reticulum, a mitochondrial unfolded protein response may play a role in the apoptotic effects of hsp90 and proteasome inhibitors.

  11. Could Upregulated Hsp70 Protein Compensate for the Hsp90-Silence-Induced Cell Death in Glioma Cells?

    Directory of Open Access Journals (Sweden)

    Chinmay Munje


    Full Text Available The molecular chaperone heat shock protein 90 alpha (Hsp90α has been recognized in various tumours including glioma. This pilot study using a proteomic approach analyses the downstream effects of Hsp90 inhibition using 17-allylamino-17-demethoxygeldanamycin (17AAG and a short hairpin RNA (shRNA oligonucleotide targeting hsp90α (shhsp90α in the U87-MG glioma cell line. Preliminary data coupled with bioinformatic analysis identified several known and unknown Hsp90 client proteins that demonstrated a change in their protein expression after Hsp90 inhibition, signifying an alteration in the canonical pathways of cell cycle progression, apoptosis, cell invasion, angiogenesis, and metastasis. Members of the glycolysis pathway were upregulated, demonstrating increased dependency on glycolysis for energy source by the treated glioma cells. Upregulated proteins also include Hsp70 and members of its family such as Hsp27 and gp96, thereby suggesting the role of Hsp90 co-chaperones in compensating for Hsp90 function after Hsp90 inhibition. Considering Hsp70’s role in antiapoptosis, it was postulated that a combination therapy involving a multitarget approach could be carried out. Consequently inhibition of both Hsp90 and Hsp70 in U87-MG glioma cells resulted in 60% cell death indicating the importance of combination therapy for glioma therapeutics.

  12. Molecular mechanisms of canalization: Hsp90 and beyond

    Indian Academy of Sciences (India)

    Neeraj Salathia; Christine Queitsch


    The Hsp90 chaperone machine facilitates the maturation of a diverse set of ‘client’ proteins. Many of these Hsp90 clients are essential nodes in signal transduction pathways and regulatory circuits, accounting for the important role Hsp90 plays in organismal development and responses to the environment. Recent findings suggest a broader impact of the chaperone on phenotype: fully functional Hsp90 canalizes wild-type phenotypes by suppressing underlying genetic and epigenetic variation. This variation can be expressed upon challenging the Hsp90 machinery by environmental stress, genetic or pharmaceutical targeting of Hsp90. The existence of Hsp90-buffered genetic and epigenetic variation together with plausible release mechanisms has wide-ranging implication for phenotype and possibly evolutionary processes. Here, we discuss the role of Hsp90 in canalization and organismal plasticity, and highlight important questions for future experimental inquiry.

  13. Applications of the HSP-matrix

    DEFF Research Database (Denmark)

    Bjarnø, Ole-Christian


    In this paper different applications of the HSP matrix are discussed. The HSP High Speed Product Management)is a new management model in which dimensions related to organisation, technology, product and market are integrated to create synergy and focus in relation to faster new product developmen....... It is shown, that the model can be used as an extension of the classic product portfolio tools, among which the most well-known are the boston Consulting Group's and McKinsey & Co.'s versions....

  14. Selection of the most appropriate two-dosemeter algorithm for estimating effective dose equivalent during maintenance periods in Korean nuclear power plants. (United States)

    Kim, Hee Geun; Kong, Tae Young


    The application of a two-dosemeter system with its algorithm, as well as a test of its use in an inhomogeneous high-radiation field, is described in this study. The goal was to improve the method for estimating the effective dose equivalent during maintenance periods at Korean Nuclear Power Plants (NPPs). The use of this method in Korean and international NPPs, including those NPPs in the USA and Canada, was also investigated. The algorithms used by the the American National Standards Institute, Lakshmanan, the National Council on Radiation Protection and Measurements (NCRP), the Electric Power Research Institute and Kim were extensively analysed as two-dosemeter algorithms. Their possible application to NPPs was evaluated using data for each algorithm from two-dosemeter results that were obtained from an inhomogeneous high-radiation field during maintenance periods at Korean NPPs. The NCRP algorithm (55:50) was selected as an optimal two-dosemeter algorithm for Korean NPPs by taking into account the field test results and the convenience of wearing two dosemeters.

  15. Active personal dosemeters in interventional radiology: tests in laboratory conditions and in hospitals. (United States)

    Clairand, I; Bordy, J-M; Daures, J; Debroas, J; Denozière, M; Donadille, L; Ginjaume, M; Itié, C; Koukorava, C; Krim, S; Lebacq, A-L; Martin, P; Struelens, L; Sans-Mercé, M; Tosic, M; Vanhavere, F


    The work package 3 of the ORAMED project, Collaborative Project (2008-11) supported by the European Commission within its seventh Framework Programme, is focused on the optimisation of the use of active personal dosemeters (APDs) in interventional radiology and cardiology (IR/IC). Indeed, a lack of appropriate APD devices is identified for these specific fields. Few devices can detect low-energy X rays (20-100 keV), and none of them are specifically designed for working in pulsed radiation fields. The work presented in this paper consists in studying the behaviour of some selected APDs deemed suitable for application in IR/IC. For this purpose, measurements under laboratory conditions, both with continuous and pulsed X-ray beams, and tests in real conditions on site in different European hospitals were performed. This study highlights the limitations of APDs for this application and the need of improving the APD technology so as to fulfil all needs in the IR/IC field.

  16. Study on the response of thermoluminescent dosemeters to synchrotron radiation: experimental method and Monte Carlo calculations. (United States)

    Bakshi, A K; Chatterjee, S; Palani Selvam, T; Dhabekar, B S


    In the present study, the energy dependence of response of some popular thermoluminescent dosemeters (TLDs) have been investigated such as LiF:Mg,Ti, LiF:Mg,Cu,P and CaSO(4):Dy to synchrotron radiation in the energy range of 10-34 keV. The study utilised experimental, Monte Carlo and analytical methods. The Monte Carlo calculations were based on the EGSnrc and FLUKA codes. The calculated energy response of all the TLDs using the EGSnrc and FLUKA codes shows excellent agreement with each other. The analytically calculated response shows good agreement with the Monte Carlo calculated response in the low-energy region. In the case of CaSO(4):Dy, the Monte Carlo-calculated energy response is smaller by a factor of 3 at all energies in comparison with the experimental response when polytetrafluoroethylene (PTFE) (75 % by wt) is included in the Monte Carlo calculations. When PTFE is ignored in the Monte Carlo calculations, the difference between the calculated and experimental response decreases (both responses are comparable >25 keV). For the LiF-based TLDs, the Monte Carlo-based response shows reasonable agreement with the experimental response.


    Figel, Markus; Stadtmann, Hannes; Grimbergen, Tom W M; McWhan, Andrew; Romero, Ana M


    Starting in 2008 the European Dosimetry Group (EURADOS) has been performing international intercomparisons on photon whole-body dosemeters for individual monitoring services. These intercomparisons were organised (on a biannual basis) in 2008, 2010, 2012 and 2014, each time with a similar set-up but with small alterations in the subsequent irradiation plans. With an increasing number of participants and participating systems, this intercomparison action has become an important tool for individual monitoring services to test their whole-body dosimetry systems, compare their results with other services or systems and to improve the quality of their dosimetry. The paper presents and compares the results of these four intercomparisons and compares the dosimetric results for the participating system types. Major dosimetric problems of the individual monitoring services are identified, and trends in the dosimetric performance of the different systems are shown. This gives the opportunity to identify some dosimetry issues that should be improved by application of the monitoring services' quality assurance systems and QA procedures.

  18. Different immunohistochemical levels of Hsp60 and Hsp70 in a subset of brain tumors and putative role of Hsp60 in neuroepithelial tumorigenesis

    Directory of Open Access Journals (Sweden)

    F. Rappa


    Full Text Available In this work we analysed, by immunohistochemistry, a series of brain tumors to detect the levels and cellular distribution of Hsp60 and Hsp70. We found that Hsp60 levels were significantly higher than those of Hsp70 in neuroepithelial tumors, while levels of both molecules were not significantly different from each other in meningeal neoplasms. In particular, Hsp60 immunopositivity was present mainly at the cytoplasmic level, while Hsp70 immunopositivity was found both in the cytoplasm and in the nucleus of tumor cells. The levels of these molecules in healthy control cells were always very low. Finally, Hsp60 and Hsp70 levels did not correlate with the different types (WHO grade of neoplasm. Our results are partially in agreement with previous studies and suggest that Hsp60 is not increased by a passive phenomenon (e.g., due to the stress caused by the peritumor environment on cancer cells but may be actively implicated in tumor progression, e.g. inhibiting tumor cell death or antitumor immune system response, as already postulated in vitro. We also briefly discuss the most recent publications on the extramitochondrial localization of Hsp60 in tumor cells and its role in tumor progression.

  19. Nuclear HSP90 and HSP70 in COPD patients treated with formoterol or formoterol and corticosteroids

    Directory of Open Access Journals (Sweden)

    Holownia A


    Full Text Available Abstract Objective Heat shock proteins assist cellular protein folding and are required for the normal activity of steroid receptors. In this study we assessed nuclear HSP90 and HSP70 proteins and mRNA levels in cells isolated from induced sputum of chronic obstructive pulmonary disease patients treated for 4 weeks with formoterol (F or formoterol+budesonide (F/ICS. Methods Nuclear heat shock protein levels were assessed by Western blot and specific mRNAs were quantified in cell lysates using qRT-PCR. Results Both HSP90 and HSP70 protein levels were higher in the F/ICS-treated patients in comparison with the F-treated group (by 31%, P Conclusions It is possible that increased nuclear heat shock proteins may play a role in the attenuation of the response to glucocorticoids in COPD patients.

  20. Altered Cross-linking of HSP27 by Zerumbone as a Novel Strategy for Overcoming HSP27- mediated Resistance

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Seo Hyun; Lee, Yoon Jin; Lee, Hae June; Lee, Yun Sil [Korea Institue of Radiological and Medical Sciences, Seoul (Korea, Republic of); Kim, Joon [Korea University, Seoul (Korea, Republic of); Seo, Woo Duck [National Institute of Crop Science, Miryang (Korea, Republic of); Nam, Joo Won; Lee, Yoo Jin; Seo, Eun Kyung [Ewha Womans University, Seoul (Korea, Republic of)


    HSPs have diverse roles in the regulation of signal transduction and in numerous aspects of cell growth and death. Indeed, HSP90, HSP70, and HSP27 have each been implicated in promoting cancer. Most HSP27 exists as large oligomeric complexes ranging from 100- 800 kDa, which are probably stabilized by complex interactions between dimeric building blocks. The functional properties of HSP27 are dependent on the quaternary structure of the protein. For example, HSP27 acts as a chaperone and binds to cytochrome c or Daxx as a dimer. Therefore, the oligomerization pattern of HPS27 is believed to have HSP27-mediated protective functions. In this study, zerumbone (ZER), the cytotoxic component isolated from Zingiber zerumbet Smith, induced cross-linking of HSP27 protein by its insertion between the disulfide bond of HSP27, and ZERmediated altered cross-linking of HSP27 modified normal HSP27 dimerization, which resulted in a sensitizing effect to tumors after treatment with radiation. Therefore, altered cross-linking by ZER may be a novel strategy for inhibition of HSP27-mediated resistance

  1. Effect of the energy dependence of response of neutron personal dosemeters routinely used in the UK on the accuracy of dose estimation

    CERN Document Server

    Tanner, R J; Thomas, D J


    A large set of neutron energy distributions have been classified by workplace to provide a guide to the neutron fields to which workers in particular industries are likely to be exposed. These have been combined (folded) with the results of a major programme of neutron personal dosemeter response function measurements, to provide results for the systematic errors that those dosemeters would give in workplaces. Data for neutron doses recorded for UK classified workers have been taken from the CIDI tables, and related to the results from the folding process. It has hence been possible to draw conclusions about the probable systematic errors that result from the use of the currently available neutron personal dosemeters, which have inherent problems associated with their energy dependence of response.

  2. Expression of Helicobacter pylori Hsp60 protein and its immunogenicity

    Institute of Scientific and Technical Information of China (English)

    Yang Bai; Liang-Ren Li; Ji-De Wang; Ye Chen; Jian-Feng Jin; Zhao-Shan Zhang; Dian-Yuan Zhou; Ya-Li Zhang


    AIM: To express Hsp60 protein of H pylori by a constructed vector and to evaluate its immunogenicity.METHODS: Hsp60 DNA was amplified by PCR and inserted into the prokaryotie expression vector pET-22b (+), which was transformed into BL21 (DE3) E. coli strain to express recombinant protein. Immunogenicity of expressed Hsp60 protein was evaluated with animal experiments.RESULTS: DNA sequence analysis showed Hsp60 DNA was the same as GenBank's research. Hsp60 recombinant protein accounted for 27.2 % of the total bacterial protein,and could be recognized by the serum from H pylori infected patients and Balb/c mice immunized with Hsp60 itself.CONCLUSION: Hsp60 recombinant protein might become a potential vaccine for controlling and treating H pylori infection.

  3. Study of teflon pads as high doses dosemeters; Estudo de pastilhas de teflon como dosimetros de doses altas

    Energy Technology Data Exchange (ETDEWEB)

    Teixeira, Maria Ines; Caldas, Linda V.E., E-mail:, E-mail: [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    The aim of this work is to study the Teflon, which is used as a binder in the manufacture of dosimetric tablets, for the feasibility of this material as high dose dosemeter. In this paper we used the technique of thermally stimulated luminescence (OSL) to characterize the dosimetric properties of Teflon. Teflon samples were exposed to different doses of radiation, using a source of gamma radiation ({sup 60}Co). It was obtained dose-response curve between 100 Gy to 50 kGy and reproducibility of OSL response. The preliminary results show that Teflon is a useful material to high dose dosimetry.

  4. Characterization of the Cardiac Overexpression of HSPB2 Reveals Mitochondrial and Myogenic Roles Supported by a Cardiac HspB2 Interactome.

    Directory of Open Access Journals (Sweden)

    Julianne H Grose

    Full Text Available Small Heat Shock Proteins (sHSPs are molecular chaperones that transiently interact with other proteins, thereby assisting with quality control of proper protein folding and/or degradation. They are also recruited to protect cells from a variety of stresses in response to extreme heat, heavy metals, and oxidative-reductive stress. Although ten human sHSPs have been identified, their likely diverse biological functions remain an enigma in health and disease, and much less is known about non-redundant roles in selective cells and tissues. Herein, we set out to comprehensively characterize the cardiac-restricted Heat Shock Protein B-2 (HspB2, which exhibited ischemic cardioprotection in transgenic overexpressing mice including reduced infarct size and maintenance of ATP levels. Global yeast two-hybrid analysis using HspB2 (bait and a human cardiac library (prey coupled with co-immunoprecipitation studies for mitochondrial target validation revealed the first HspB2 "cardiac interactome" to contain many myofibril and mitochondrial-binding partners consistent with the overexpression phenotype. This interactome has been submitted to the Biological General Repository for Interaction Datasets (BioGRID. A related sHSP chaperone HspB5 had only partially overlapping binding partners, supporting specificity of the interactome as well as non-redundant roles reported for these sHSPs. Evidence that the cardiac yeast two-hybrid HspB2 interactome targets resident mitochondrial client proteins is consistent with the role of HspB2 in maintaining ATP levels and suggests new chaperone-dependent functions for metabolic homeostasis. One of the HspB2 targets, glyceraldehyde 3-phosphate dehydrogenase (GAPDH, has reported roles in HspB2 associated phenotypes including cardiac ATP production, mitochondrial function, and apoptosis, and was validated as a potential client protein of HspB2 through chaperone assays. From the clientele and phenotypes identified herein, it is

  5. The Anti-inflammatory mechanisms of Hsp70

    Directory of Open Access Journals (Sweden)

    Thiago J Borges


    Full Text Available Immune responses to heat shock proteins (Hsp develop in virtually all inflammatory diseases; however, the significance of such responses is only now becoming clear. In experimental disease models, Hsp administration can prevent or arrest inflammatory damage, and in initial clinical trials in patients with chronic inflammatory diseases, Hsp peptides have been shown to promote the production of anti-inflammatory cytokines, indicating immunoregulatory potential of Hsp. Therefore, the presence of immune responses to Hsp in inflammatory diseases can be seen as an attempt of the immune system to correct the inflammatory condition. Hsp70 can modulate inflammatory responses in models of arthritis, colitis and graft rejection, and the mechanisms underlying this effect are now being elucidated. Incubation with microbial Hsp70 was seen to induce tolerogenic DCs and to promote a suppressive phenotype in myeloid-derived suppressor cells and monocytes. These DC could induce regulatory T cells (Tregs, independently of the antigens they presented. Some Hsp70 family members are associated with autophagy, leading to a preferential uploading of Hsp70 peptides in MHC class II molecules of stressed cells. Henceforth, conserved Hsp70 peptides may be presented in these situations and constitute targets of Tregs, contributing to downregulation of inflammation. Finally, an interfering effect in multiple intracellular inflammatory signaling pathways is also known for Hsp70. Altogether it seems attractive to use Hsp70, or its derivative peptides, for modulation of inflammation. This is a physiological immunotherapy approach, without the immediate necessity of defining disease specific auto-antigens. In this article, we present the evidence on anti-inflammatory effects of Hsp70 and discuss the need for experiments that will be crucial for the further exploration of the immuno-suppressive potential of this protein.

  6. Reconstructing the Hsp90/Tau Machine. (United States)

    Jinwal, Umesh K; Koren, John; Dickey, Chad A


    Imbalanced protein load within cells is a critical aspect for most diseases of aging. In particular, the accumulation of proteins into neurotoxic aggregates is a common thread for a host of neurodegenerative diseases. Recent work demonstrates that age-related changes to the cellular chaperone repertoire contributes to abnormal buildup of the microtubule-associated protein tau that accumulates in a group of diseases termed tauopathies, the most common being Alzheimer's disease (AD). The Hsp90 co-chaperone repertoire has diverse effects on tau stability; some co-chaperones stabilize tau while others facilitate its clearance. We propose that each of these proteins may be novel therapeutic targets. While targeting Hsp90 directly may be deleterious at the organismal level, perhaps targeting individual co-chaperone activities will be more tolerable.

  7. Hsp70/Hsp90 organising protein (hop): beyond interactions with chaperones and prion proteins. (United States)

    Baindur-Hudson, Swati; Edkins, Adrienne L; Blatch, Gregory L


    The Hsp70/Hsp90 organising protein (Hop), also known as stress-inducible protein 1 (STI1), has received considerable attention for diverse cellular functions in both healthy and diseased states. There is extensive evidence that intracellular Hop is a co-chaperone of the major chaperones Hsp70 and Hsp90, playing an important role in the productive folding of Hsp90 client proteins. Consequently, Hop is implicated in a number of key signalling pathways, including aberrant pathways leading to cancer. However, Hop is also secreted and it is now well established that Hop also serves as a receptor for the prion protein, PrP(C). The intracellular and extracellular forms of Hop most likely represent two different isoforms, although the molecular determinants of these divergent functions are yet to be identified. There is also a growing body of research that reports the involvement of Hop in cellular activities that appear independent of either chaperones or PrP(C). While Hop has been shown to have various cellular functions, its biological function remains elusive. However, recent knockout studies in mammals suggest that Hop has an important role in embryonic development. This review provides a critical overview of the latest molecular, cellular and biological research on Hop, critically evaluating its function in healthy systems and how this function is adapted in diseases states.

  8. Imaging of Hsp70-positive tumors with cmHsp70.1 antibody-conjugated gold nanoparticles. (United States)

    Gehrmann, Mathias K; Kimm, Melanie A; Stangl, Stefan; Schmid, Thomas E; Noël, Peter B; Rummeny, Ernst J; Multhoff, Gabriele


    Real-time imaging of small tumors is still one of the challenges in cancer diagnosis, prognosis, and monitoring of clinical outcome. Targeting novel biomarkers that are selectively expressed on a large variety of different tumors but not normal cells has the potential to improve the imaging capacity of existing methods such as computed tomography. Herein, we present a novel technique using cmHsp70.1 monoclonal antibody-conjugated spherical gold nanoparticles for quantification of the targeted uptake of gold nanoparticles into membrane Hsp70-positive tumor cells. Upon binding, cmHsp70.1-conjugated gold nanoparticles but not nanoparticles coupled to an isotype-matched IgG1 antibody or empty nanoparticles are rapidly taken up by highly malignant Hsp70 membrane-positive mouse tumor cells. After 24 hours, the cmHsp70.1-conjugated gold nanoparticles are found to be enriched in the perinuclear region. Specificity for membrane Hsp70 was shown by using an Hsp70 knockout tumor cell system. Toxic side effects of the cmHsp70.1-conjugated nanoparticles are not observed at a concentration of 1-10 µg/mL. Experiments are ongoing to evaluate whether cmHsp70.1 antibody-conjugated gold nanoparticles are suitable for the detection of membrane-Hsp70-positive tumors in vivo.

  9. Mps1 Mediated Phosphorylation of Hsp90 Confers Renal Cell Carcinoma Sensitivity and Selectivity to Hsp90 Inhibitors

    Directory of Open Access Journals (Sweden)

    Mark R. Woodford


    Full Text Available The molecular chaperone Hsp90 protects deregulated signaling proteins that are vital for tumor growth and survival. Tumors generally display sensitivity and selectivity toward Hsp90 inhibitors; however, the molecular mechanism underlying this phenotype remains undefined. We report that the mitotic checkpoint kinase Mps1 phosphorylates a conserved threonine residue in the amino-domain of Hsp90. This, in turn, regulates chaperone function by reducing Hsp90 ATPase activity while fostering Hsp90 association with kinase clients, including Mps1. Phosphorylation of Hsp90 is also essential for the mitotic checkpoint because it confers Mps1 stability and activity. We identified Cdc14 as the phosphatase that dephosphorylates Hsp90 and disrupts its interaction with Mps1. This causes Mps1 degradation, thus providing a mechanism for its inactivation. Finally, Hsp90 phosphorylation sensitizes cells to its inhibitors, and elevated Mps1 levels confer renal cell carcinoma selectivity to Hsp90 drugs. Mps1 expression level can potentially serve as a predictive indicator of tumor response to Hsp90 inhibitors.

  10. The Malarial Exported PFA0660w Is an Hsp40 Co-Chaperone of PfHsp70-x.

    Directory of Open Access Journals (Sweden)

    Michael O Daniyan

    Full Text Available Plasmodium falciparum, the human pathogen responsible for the most dangerous malaria infection, survives and develops in mature erythrocytes through the export of proteins needed for remodelling of the host cell. Molecular chaperones of the heat shock protein (Hsp family are prominent members of the exportome, including a number of Hsp40s and a Hsp70. PFA0660w, a type II Hsp40, has been shown to be exported and possibly form a complex with PfHsp70-x in the infected erythrocyte cytosol. However, the chaperone properties of PFA0660w and its interaction with human and parasite Hsp70s are yet to be investigated. Recombinant PFA0660w was found to exist as a monomer in solution, and was able to significantly stimulate the ATPase activity of PfHsp70-x but not that of a second plasmodial Hsp70 (PfHsp70-1 or a human Hsp70 (HSPA1A, indicating a potential specific functional partnership with PfHsp70-x. Protein binding studies in the presence and absence of ATP suggested that the interaction of PFA0660w with PfHsp70-x most likely represented a co-chaperone/chaperone interaction. Also, PFA0660w alone produced a concentration-dependent suppression of rhodanese aggregation, demonstrating its chaperone properties. Overall, we have provided the first biochemical evidence for the possible role of PFA0660w as a chaperone and as co-chaperone of PfHsp70-x. We propose that these chaperones boost the chaperone power of the infected erythrocyte, enabling successful protein trafficking and folding, and thereby making a fundamental contribution to the pathology of malaria.

  11. Hsp70 and the Cochaperone StiA (Hop) Orchestrate Hsp90-Mediated Caspofungin Tolerance in Aspergillus fumigatus. (United States)

    Lamoth, Frédéric; Juvvadi, Praveen R; Soderblom, Erik J; Moseley, M Arthur; Steinbach, William J


    Aspergillus fumigatus is the primary etiologic agent of invasive aspergillosis (IA), a major cause of death among immunosuppressed patients. Echinocandins (e.g., caspofungin) are increasingly used as second-line therapy for IA, but their activity is only fungistatic. Heat shock protein 90 (Hsp90) was previously shown to trigger tolerance to caspofungin and the paradoxical effect (i.e., decreased efficacy of caspofungin at higher concentrations). Here, we demonstrate the key role of another molecular chaperone, Hsp70, in governing the stress response to caspofungin via Hsp90 and their cochaperone Hop/Sti1 (StiA in A. fumigatus). Mutation of the StiA-interacting domain of Hsp70 (C-terminal EELD motif) impaired thermal adaptation and caspofungin tolerance with loss of the caspofungin paradoxical effect. Impaired Hsp90 function and increased susceptibility to caspofungin were also observed following pharmacologic inhibition of the C-terminal domain of Hsp70 by pifithrin-μ or after stiA deletion, further supporting the links among Hsp70, StiA, and Hsp90 in governing caspofungin tolerance. StiA was not required for the physical interaction between Hsp70 and Hsp90 but had distinct roles in the regulation of their function in caspofungin and heat stress responses. In conclusion, this study deciphering the physical and functional interactions of the Hsp70-StiA-Hsp90 complex provided new insights into the mechanisms of tolerance to caspofungin in A. fumigatus and revealed a key C-terminal motif of Hsp70, which can be targeted by specific inhibitors, such as pifithrin-μ, to enhance the antifungal activity of caspofungin against A. fumigatus.

  12. Molecular characterization of the Corynebacterium pseudotuberculosis hsp60-hsp10 operon, and evaluation of the immune response and protective efficacy induced by hsp60 DNA vaccination in mice

    Directory of Open Access Journals (Sweden)

    Oliveira Sérgio C


    Full Text Available Abstract Background Heat shock proteins (HSPs are important candidates for the development of vaccines because they are usually able to promote both humoral and cellular immune responses in mammals. We identified and characterized the hsp60-hsp10 bicistronic operon of the animal pathogen Corynebacterium pseudotuberculosis, a Gram-positive bacterium of the class Actinobacteria, which causes caseous lymphadenitis (CLA in small ruminants. Findings To construct the DNA vaccine, the hsp60 gene of C. pseudotuberculosis was cloned in a mammalian expression vector. BALB/c mice were immunized by intramuscular injection with the recombinant plasmid (pVAX1/hsp60. Conclusion This vaccination induced significant anti-hsp60 IgG, IgG1 and IgG2a isotype production. However, immunization with this DNA vaccine did not confer protective immunity.

  13. The Complex Function of Hsp70 in Metastatic Cancer

    Directory of Open Access Journals (Sweden)

    Kata Juhasz


    Full Text Available Elevated expression of the inducible heat shock protein 70 (Hsp70 is known to correlate with poor prognosis in many cancers. Hsp70 confers survival advantage as well as resistance to chemotherapeutic agents, and promotes tumor cell invasion. At the same time, tumor-derived extracellular Hsp70 has been recognized as a “chaperokine”, activating antitumor immunity. In this review we discuss localization dependent functions of Hsp70 in the context of invasive cancer. Understanding the molecular principles of metastasis formation steps, as well as interactions of the tumor cells with the microenvironment and the immune system is essential for fighting metastatic cancer. Although Hsp70 has been implicated in different steps of the metastatic process, the exact mechanisms of its action remain to be explored. Known and potential functions of Hsp70 in controlling or modulating of invasion and metastasis are discussed.

  14. The Complex Function of Hsp70 in Metastatic Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Juhasz, Kata; Lipp, Anna-Maria; Nimmervoll, Benedikt; Sonnleitner, Alois; Hesse, Jan; Haselgruebler, Thomas; Balogi, Zsolt, E-mail: [Center for Advanced Bioanalysis GmbH, Gruberstr. 40-42, A-4020 Linz (Austria)


    Elevated expression of the inducible heat shock protein 70 (Hsp70) is known to correlate with poor prognosis in many cancers. Hsp70 confers survival advantage as well as resistance to chemotherapeutic agents, and promotes tumor cell invasion. At the same time, tumor-derived extracellular Hsp70 has been recognized as a “chaperokine”, activating antitumor immunity. In this review we discuss localization dependent functions of Hsp70 in the context of invasive cancer. Understanding the molecular principles of metastasis formation steps, as well as interactions of the tumor cells with the microenvironment and the immune system is essential for fighting metastatic cancer. Although Hsp70 has been implicated in different steps of the metastatic process, the exact mechanisms of its action remain to be explored. Known and potential functions of Hsp70 in controlling or modulating of invasion and metastasis are discussed.

  15. Functional and structural analysis of maize hsp101 IRES.

    Directory of Open Access Journals (Sweden)

    Augusto Samuel Jiménez-González

    Full Text Available Maize heat shock protein of 101 KDa (HSP101 is essential for thermotolerance induction in this plant. The mRNA encoding this protein harbors an IRES element in the 5'UTR that mediates cap-independent translation initiation. In the current work it is demonstrated that hsp101 IRES comprises the entire 5'UTR sequence (150 nts, since deletion of 17 nucleotides from the 5' end decreased translation efficiency by 87% compared to the control sequence. RNA structure analysis of maize hsp101 IRES revealed the presence of three stem-loops toward its 5' end, whereas the remainder sequence contains a great proportion of unpaired nucleotides. Furthermore, HSP90 protein was identified by mass spectrometry as the protein preferentially associated with the maize hsp101 IRES. In addition, it has been found that eIFiso4G rather than eIF4G initiation factor mediates translation of the maize hsp101 mRNA.

  16. About the measurements systems with pen and thermoluminescent dosemeters; Sobre los sistemas de medicion con dosimetros de pluma y los termoluminiscentes

    Energy Technology Data Exchange (ETDEWEB)

    Cortes I, M.E.; Ramirez G, F.P. [Instituto Mexicano del Petroleo, Eje Central Lazaro Cardenas 152. 07730 Mexico D.F. (Mexico)


    In this work it is presented dosimetric data obtained with pen and thermoluminescent dosemeters, which are used by the Occupational Exposure Personnel (OEP) of the Mexican Petroleum Institute (IMP)(1). It was marked several great characteristics as for example, the differences among units which use one and another dosemeter type. Likewise, it is given to know diverse problems that were had in the IMP at relating the data obtained with these dosemeters (which utilizes OEP) and the ICRP 60 recommendations 1990. One of the most important difficulties is to satisfy the recommended limits by ICRP, particularly those that are referring to the units and their complex calculations. With respect to the unities, the ICRP makes reference at the concepts `dose equivalent` and `effective dose` with the sievert unit, that the General Regulations for Radiological Safety associates with `dose equivalent` and `effective dose equivalent`. It was illustrated the type of dosimetric statistics which are obtained with the TLD lectures and a OEP pen dosemeter during 1997. (Author)

  17. Hsp90 governs dispersion and drug resistance of fungal biofilms.

    Directory of Open Access Journals (Sweden)

    Nicole Robbins


    Full Text Available Fungal biofilms are a major cause of human mortality and are recalcitrant to most treatments due to intrinsic drug resistance. These complex communities of multiple cell types form on indwelling medical devices and their eradication often requires surgical removal of infected devices. Here we implicate the molecular chaperone Hsp90 as a key regulator of biofilm dispersion and drug resistance. We previously established that in the leading human fungal pathogen, Candida albicans, Hsp90 enables the emergence and maintenance of drug resistance in planktonic conditions by stabilizing the protein phosphatase calcineurin and MAPK Mkc1. Hsp90 also regulates temperature-dependent C. albicans morphogenesis through repression of cAMP-PKA signalling. Here we demonstrate that genetic depletion of Hsp90 reduced C. albicans biofilm growth and maturation in vitro and impaired dispersal of biofilm cells. Further, compromising Hsp90 function in vitro abrogated resistance of C. albicans biofilms to the most widely deployed class of antifungal drugs, the azoles. Depletion of Hsp90 led to reduction of calcineurin and Mkc1 in planktonic but not biofilm conditions, suggesting that Hsp90 regulates drug resistance through different mechanisms in these distinct cellular states. Reduction of Hsp90 levels led to a marked decrease in matrix glucan levels, providing a compelling mechanism through which Hsp90 might regulate biofilm azole resistance. Impairment of Hsp90 function genetically or pharmacologically transformed fluconazole from ineffectual to highly effective in eradicating biofilms in a rat venous catheter infection model. Finally, inhibition of Hsp90 reduced resistance of biofilms of the most lethal mould, Aspergillus fumigatus, to the newest class of antifungals to reach the clinic, the echinocandins. Thus, we establish a novel mechanism regulating biofilm drug resistance and dispersion and that targeting Hsp90 provides a much-needed strategy for improving

  18. Hsp60 response in experimental and human temporal lobe epilepsy. (United States)

    Marino Gammazza, Antonella; Colangeli, Roberto; Orban, Gergely; Pierucci, Massimo; Di Gennaro, Giancarlo; Lo Bello, Margherita; D'Aniello, Alfredo; Bucchieri, Fabio; Pomara, Cristoforo; Valentino, Mario; Muscat, Richard; Benigno, Arcangelo; Zummo, Giovanni; de Macario, Everly Conway; Cappello, Francesco; Di Giovanni, Giuseppe; Macario, Alberto J L


    The mitochondrial chaperonin Hsp60 is a ubiquitous molecule with multiple roles, constitutively expressed and inducible by oxidative stress. In the brain, Hsp60 is widely distributed and has been implicated in neurological disorders, including epilepsy. A role for mitochondria and oxidative stress has been proposed in epileptogenesis of temporal lobe epilepsy (TLE). Here, we investigated the involvement of Hsp60 in TLE using animal and human samples. Hsp60 immunoreactivity in the hippocampus, measured by Western blotting and immunohistochemistry, was increased in a rat model of TLE. Hsp60 was also increased in the hippocampal dentate gyrus neurons somata and neuropil and hippocampus proper (CA3, CA1) of the epileptic rats. We also determined the circulating levels of Hsp60 in epileptic animals and TLE patients using ELISA. The epileptic rats showed circulating levels of Hsp60 higher than controls. Likewise, plasma post-seizure Hsp60 levels in patients were higher than before the seizure and those of controls. These results demonstrate that Hsp60 is increased in both animals and patients with TLE in affected tissues, and in plasma in response to epileptic seizures, and point to it as biomarker of hippocampal stress potentially useful for diagnosis and patient management.

  19. Development of Noviomimetics as C-Terminal Hsp90 Inhibitors. (United States)

    Anyika, Mercy; McMullen, Mason; Forsberg, Leah K; Dobrowsky, Rick T; Blagg, Brian S J


    KU-32 and KU-596 are novobiocin-derived, C-terminal heat shock protein 90 (Hsp90) modulators that induce Hsp70 levels and manifest neuroprotective activity. However, the synthetically complex noviose sugar requires 10 steps to prepare, which makes translational development difficult. In this study, we developed a series of "noviomimetic" analogues of KU-596, which contain noviose surrogates that can be easily prepared, while maintaining the ability to induce Hsp70 levels. Both sugar and sugar analogues were designed, synthesized, and evaluated in a luciferase reporter assay, which identified compound 37, a benzyl containing noviomimetic, as the most potent inducer of Hsp70.

  20. ANTI-HSP60 and ANTI-HSP70 antibody levels and micro/ macrovascular complications in type 1 diabetes: the EURODIAB Study

    DEFF Research Database (Denmark)

    Gruden, G.; Bruno, G.; Chaturvedi, N.;


    OBJECTIVES: The heat shock proteins 60 and 70 (HSP60, HSP70) play an important role in cytoprotection. Under stress conditions they are released into the circulation and elicit an immune response. Anti-HSP60 and anti-HSP70 antibody levels have been associated with cardiovascular disease. Type 1...

  1. Multi-layered molecular mechanisms of polypeptide holding, unfolding and disaggregation by HSP70/HSP110 chaperones (United States)

    Finka, Andrija; Sharma, Sandeep K.; Goloubinoff, Pierre


    Members of the HSP70/HSP110 family (HSP70s) form a central hub of the chaperone network controlling all aspects of proteostasis in bacteria and the ATP-containing compartments of eukaryotic cells. The heat-inducible form HSP70 (HSPA1A) and its major cognates, cytosolic HSC70 (HSPA8), endoplasmic reticulum BIP (HSPA5), mitochondrial mHSP70 (HSPA9) and related HSP110s (HSPHs), contribute about 3% of the total protein mass of human cells. The HSP70s carry out a plethora of housekeeping cellular functions, such as assisting proper de novo folding, assembly and disassembly of protein complexes, pulling polypeptides out of the ribosome and across membrane pores, activating and inactivating signaling proteins and controlling their degradation. The HSP70s can induce structural changes in alternatively folded protein conformers, such as clathrin cages, hormone receptors and transcription factors, thereby regulating vesicular trafficking, hormone signaling and cell differentiation in development and cancer. To carry so diverse cellular housekeeping and stress-related functions, the HSP70s act as ATP-fuelled unfolding nanomachines capable of switching polypeptides between different folded states. During stress, the HSP70s can bind (hold) and prevent the aggregation of misfolding proteins and thereafter act alone or in collaboration with other unfolding chaperones to solubilize protein aggregates. Here, we discuss the common ATP-dependent mechanisms of holding, unfolding-by-clamping and unfolding-by-entropic pulling, by which the HSP70s can apparently convert various alternatively folded and misfolded polypeptides into differently active conformers. Understanding how HSP70s can prevent the formation of cytotoxic protein aggregates, pull, unfold, and solubilize them into harmless species is central to the design of therapies against protein conformational diseases. PMID:26097841

  2. C-terminal sequences of hsp70 and hsp90 as non-specific anchors for tetratricopeptide repeat (TPR) proteins. (United States)

    Ramsey, Andrew J; Russell, Lance C; Chinkers, Michael


    Steroid-hormone-receptor maturation is a multi-step process that involves several TPR (tetratricopeptide repeat) proteins that bind to the maturation complex via the C-termini of hsp70 (heat-shock protein 70) and hsp90 (heat-shock protein 90). We produced a random T7 peptide library to investigate the roles played by the C-termini of the two heat-shock proteins in the TPR-hsp interactions. Surprisingly, phages with the MEEVD sequence, found at the C-terminus of hsp90, were not recovered from our biopanning experiments. However, two groups of phages were isolated that bound relatively tightly to HsPP5 (Homo sapiens protein phosphatase 5) TPR. Multiple copies of phages with a C-terminal sequence of LFG were isolated. These phages bound specifically to the TPR domain of HsPP5, although mutation studies produced no evidence that they bound to the domain's hsp90-binding groove. However, the most abundant family obtained in the initial screen had an aspartate residue at the C-terminus. Two members of this family with a C-terminal sequence of VD appeared to bind with approximately the same affinity as the hsp90 C-12 control. A second generation pseudo-random phage library produced a large number of phages with an LD C-terminus. These sequences acted as hsp70 analogues and had relatively low affinities for hsp90-specific TPR domains. Unfortunately, we failed to identify residues near hsp90's C-terminus that impart binding specificity to individual hsp90-TPR interactions. The results suggest that the C-terminal sequences of hsp70 and hsp90 act primarily as non-specific anchors for TPR proteins.

  3. Disease-associated mutations in the HSPD1 gene encoding the large subunit of the mitochondrial HSP60/HSP10 chaperonin complex

    Directory of Open Access Journals (Sweden)

    Peter Bross


    Full Text Available Heat shock protein 60 (HSP60 forms together with heat shock protein 10 (HSP10 double-barrel chaperonin complexes that are essential for folding to the native state of proteins in the mitochondrial matrix space. Two extremely rare monogenic disorders have been described that are caused by missense mutations in the HSPD1 gene that encodes the HSP60 subunit of the HSP60/HSP10 chaperonin complex. Investigations of the molecular mechanisms underlying these disorders have revealed that different degrees of reduced HSP60 function produce distinct neurological phenotypes. While mutations with deleterious or strong dominant negative effects are not compatible with life, HSPD1 gene variations found in the human population impair HSP60 function and depending on the mechanism and degree of HSP60 dys- and malfunction cause different phenotypes. We here summarize the knowledge on the effects of disturbances of the function of the HSP60/HSP10 chaperonin complex by disease-associated mutations.

  4. Heat Shock Protein (HSP peptide and HSP-based immunotherapies for the treatment of cancer

    Directory of Open Access Journals (Sweden)

    Gabriele eMulthoff


    Full Text Available Intracellular residing heat shock proteins (HSPs with a molecular weight of approximately 70 and 90 kDa function as molecular chaperones that assist folding/unfolding and transport of proteins across membranes and prevent protein aggregation after environmental stress. In contrast to normal cells, tumor cells have higher cytosolic HSP70 and Hsp90 levels which contribute to tumor cell propagation, metastasis and protection against apoptosis. In addition to their intracellular chaperoning functions, extracellular localized and membrane-bound HSPs have been found to play key roles in eliciting anti-tumor immune responses either by acting as carriers for tumor-derived, immunogenic peptides, as adjuvants for antigen presentation or as targets for the innate immune system. The interaction of HSP-peptide complexes or peptide-free HSPs with receptors on antigen presenting cells (APCs promotes the maturation of dendritic cells (DCs, results in an up-regulation of MHC class I and class II molecules, induces secretion of pro- and anti-inflammatory cytokines, chemokines, and immune modulatory nitric oxides and thus integrate adaptive and innate immune phenomena. Herein, we aim to recapitulate the history and current status of HSP-based immunotherapies and vaccination strategies in the treatment of cancer.

  5. TL response of LiF: Mg, Cu, P dosemeters in function of the photon energy; Respuesta TL de dosimetros de LiF: Mg, Cu, P en funcion de la energia del foton

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez, P.R. [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico); Azorin, J. [UAM-I, 09340 Mexico D.F. (Mexico); Furetta, C. [Universita degli Studi di Roma ' La Sapienza' , Piazzale Aldo Moro 2, 00185 Roma (Italy)


    In this work the obtained results of studying the response of dosemeters of LiF: Mg, Cu, P + Ptfe, developed in the ININ, commercial dosemeters GR200A (LiF: Mg, Cu, P) of chinese manufacture and TLD-100 (LiF: Mg, Ti) US are presented, when irradiating them with X rays 16, 24, 34.5, 42, 100 and 145 keV of effective energy, with gamma rays of 662 keV ({sup 137} Cs) and 1252 keV ({sup 60} Co). The results are presented in function of the sensitivity that the dosemeters showed normalized to gamma radiation of {sup 60} Co. It was appreciated that the dosemeters more equivalent to the tissue, they were those of LiF: Mg, Cu, P + Ptfe, while the less equivalent ones were the TLD-100. (Author)

  6. Stoichiometric expression of mtHsp40 and mtHsp70 modulates mitochondrial morphology and cristae structure via Opa1L cleavage. (United States)

    Lee, Byoungchun; Ahn, Younghee; Kang, Sung-Myung; Park, Youngjin; Jeon, You-Jin; Rho, Jong M; Kim, Sung-Woo


    Deregulation of mitochondrial heat-shock protein 40 (mtHsp40) and dysfunction of mtHsp70 are associated with mitochondrial fragmentation, suggesting that mtHsp40 and mtHsp70 may play roles in modulating mitochondrial morphology. However, the mechanism of mitochondrial fragmentation induced by mtHsp40 deregulation and mtHsp70 dysfunction remains unclear. In addition, the functional link between mitochondrial morphology change upon deregulated mtHsp40/mtHsp70 and mitochondrial function has been unexplored. Our coimmunoprecipitation and protein aggregation analysis showed that both overexpression and depletion of mtHsp40 accumulated aggregated proteins in fragmented mitochondria. Moreover, mtHsp70 loss and expression of a mtHsp70 mutant lacking the client-binding domain caused mitochondrial fragmentation. Together the data suggest that the molecular ratio of mtHsp40 to mtHsp70 is important for their chaperone function and mitochondrial morphology. Whereas mitochondrial translocation of Drp1 was not altered, optic atrophy 1 (Opa1) short isoform accumulated in fragmented mitochondria, suggesting that mitochondrial fragmentation in this study results from aberration of mitochondrial inner membrane fusion. Finally, we found that fragmented mitochondria were defective in cristae development, OXPHOS, and ATP production. Taken together, our data suggest that impaired stoichiometry between mtHsp40 and mtHsp70 promotes Opa1L cleavage, leading to cristae opening, decreased OXPHOS, and triggering of mitochondrial fragmentation after reduction in their chaperone function.

  7. Comparative Study on the Immunogenicity between Hsp70 DNA Vaccine and Hsp65 DNA Vaccine in Human Mycobacterium Tuberculosis

    Institute of Scientific and Technical Information of China (English)

    DAI; Wuxing; HUANG; Hailang; YUAN; Ye; HU; Jiajie; HUANGFU; Yongmu


    The BALB/c mice were immunized with Hsp70 DNA and Hsp65 DNA vaccines in human Mycobacterium tuberculosis. Eight weeks after immunization, the eyeballs were removed, blood and spleen taken, and intraperitoneal macrophages were harvested. The lymphocytic stimulating index(SI) was used to measure the cellular proliferating ability and NO release to measure the phagocytic activity of the macrophages. With ELISA kit, the levels of interleukin-2 (IL-2) and interferon-γ(IFN-γ) in serum and the splenic lymphocytic cultured supernatant were detected. The results showed that after the mice were immunized with 100 μg/mouse of Hsp70 DNA vaccine intramuscularly, the splenic lymphocytic proliferating ability in the mice was significantly increased as compared with that in the control group, vector group and Hsp65 DNA vaccine group (P<0. 01); The contents of NO in the intraperitoneal macrophages of the mice were significantly lower than in the control group and Hsp65 DNA vaccine group (P<0. 01); The levels of serum IL-2 in the mice were significantly higher than in the control group, but there was no statistical difference between Hsp65 DNA group and vector group (P>0. 05); The contents of serum IFN-γ in the mice were significantly higher than in the control group, but significantly lower than in the Hsp65 DNA vaccine group (P<0. 05). It was indicated that immunization with Hsp70 DNA vaccine could obviously enhance the immune response, but its intensity seemed inferior to Hsp65 DNA vaccine. The anti-infection mechanisms and clinical use in the future of the vaccines of Hsp70 DNA and Hsp65 DNA are worth further studying.

  8. Expression of Hsp27 and Hsp70 in lymphocytes and plasma in healthy workers and coal miners with lung cancer. (United States)

    Wang, Haijiao; Xing, Jingcai; Wang, Feng; Han, Wenhui; Ren, Houmao; Wu, Tangchun; Chen, Weihong


    In coal mines, main occupational hazard is coal-mine dust, which can cause health problem including coal workers' pneumoconiosis and lung cancer. Some heat shock proteins (Hsps) have been reported as an acute response to a wide variety of stressful stimuli. Whether Hsps protect against chronic environmental coal-mine dust over years is unknown. It is also interesting to know that whether the expression of Hsp27 and Hsp70 proteins as a marker for exposure is associated risk of lung cancer among coal miners. We investigated the association between levels of Hsp27 and Hsp70 expression in lymphocytes and plasma and levels of coal-mine dust exposure in workplace or risk of lung cancer in 42 cancer-free non-coal miners, 99 cancer-free coal miners and 51 coal miners with lung cancer in Taiyuan city in China. The results showed that plasma Hsp27 levels were increased in coal miners compared to non-coal miners (P<0.01). Except high cumulative coal-mine dust exposure (OR=13.62, 95%CI=6.05-30.69) and amount of smoking higher than 24 pack-year (OR=2.72, 95% CI=1.37-5.42), the elevated levels of plasma Hsp70 (OR=13.00, 95% CI=5.14-32.91) and plasma Hsp27 (OR=2.97, 95% CI=1.40-6.32) and decreased expression of Hsp70 in lymphocytes (OR=2.36, 95% CI=1.05-5.31) were associated with increased risk of lung cancer. These findings suggest that plasma Hsp27 may be a potential marker for coal-mine dust exposure. And the expression of Hsp27 and Hsp70 levels in plasma and lymphocytes may be used as biomarkers for lung cancer induced by occupational coal-mine dust exposure.

  9. Hsp90 regulation of fibroblast activation in pulmonary fibrosis (United States)

    Sontake, Vishwaraj; Wang, Yunguan; Kasam, Rajesh K.; Sinner, Debora; Reddy, Geereddy B.; Naren, Anjaparavanda P.; McCormack, Francis X.; Jegga, Anil G.; Madala, Satish K.


    Idiopathic pulmonary fibrosis (IPF) is a severe fibrotic lung disease associated with fibroblast activation that includes excessive proliferation, tissue invasiveness, myofibroblast transformation, and extracellular matrix (ECM) production. To identify inhibitors that can attenuate fibroblast activation, we queried IPF gene signatures against a library of small-molecule-induced gene-expression profiles and identified Hsp90 inhibitors as potential therapeutic agents that can suppress fibroblast activation in IPF. Although Hsp90 is a molecular chaperone that regulates multiple processes involved in fibroblast activation, it has not been previously proposed as a molecular target in IPF. Here, we found elevated Hsp90 staining in lung biopsies of patients with IPF. Notably, fibroblasts isolated from fibrotic lesions showed heightened Hsp90 ATPase activity compared with normal fibroblasts. 17-N-allylamino-17-demethoxygeldanamycin (17-AAG), a small-molecule inhibitor of Hsp90 ATPase activity, attenuated fibroblast activation and also TGF-β–driven effects on fibroblast to myofibroblast transformation. The loss of the Hsp90AB, but not the Hsp90AA isoform, resulted in reduced fibroblast proliferation, myofibroblast transformation, and ECM production. Finally, in vivo therapy with 17-AAG attenuated progression of established and ongoing fibrosis in a mouse model of pulmonary fibrosis, suggesting that targeting Hsp90 represents an effective strategy for the treatment of fibrotic lung disease. PMID:28239659

  10. Use of active dosemeters as a optimization tool in hemodynamics; Uso de dosimetros ativos como ferramenta de otimizacao em hemodinamica

    Energy Technology Data Exchange (ETDEWEB)

    Nunes, Rafael; Pereira, Dirceu D. [Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ (Brazil). Inst. de Fisica; Canevaro, Lucia V. [Instituto de Radioprotecao e Dosimetria (IRD/CNEN-RJ), Rio de Janeiro, RJ (Brazil); Rodrigues, Barbara B.D. [Comissao Nacional de Energia Nuclear (CNEN), Rio de Janeiro, RJ (Brazil); Ferreira, Esmeralci [Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, RJ (Brazil). Faculdade de Medicina. Servico de Hemodinamica


    Interventional cardiology procedures are, in general, associated with high doses in patients and professionals. The objective of this study is to measure the radiation levels received by professionals .The professional dosimetry was performed in a department of Hemodynamics of University Hospital in Rio de Janeiro. were followed 331 coronary angiography (CA) and 26 percutaneous transluminal coronary angioplasty (PTCA) procedures. For this, were used active dosemeters to measure the radiation levels at the chest of interventional professionals. The results show that average personal equivalent dose of doctors, per procedure was 100 e 154 μSv. On average, nursing technicians and radiologist receive 12 and 10% of doses of physicians, respectively, during CA procedures. From the results, it appears that the doses of hemodynamics exceed the annual dose limit of the standards. The use of lead shielding is presented as an effective action to reduce doses in these workers. (author)

  11. Performance of the electronic personal dosemeter for neutron 'Saphydose-N' at different workplaces of nuclear facilities. (United States)

    Lahaye, T; Chau, Q; Ménard, S; Lacoste, V; Muller, H; Luszik-Bhadra, M; Reginatto, M; Bruguier, P


    This paper mainly aims at presenting the measurements and the results obtained with the electronic personal neutron dosemeter Saphydose-N at different facilities. Three campaigns were led in the frame of the European contract EVIDOS ('Evaluation of Individual Dosimetry in Mixed Neutron and Photon Radiation Fields'). The first one consisted in the measurements at the IRSN French research laboratory in reference neutron fields generated by a thermal facility (SIGMA), radionuclide ISO sources ((241)AmBe; (252)Cf; (252)Cf(D(2)O)\\Cd) and a realistic spectrum (CANEL/T400). The second one was performed at the Krümmel Nuclear Power Plant (Germany) close to the boiling water reactor and to a spent fuel transport cask. The third one was realised at Mol (Belgium), at the VENUS Research Reactor and at Belgonucléaire, a fuel processing factory.

  12. Absorbed dose measurements in mammography using Monte Carlo method and ZrO{sub 2}+PTFE dosemeters

    Energy Technology Data Exchange (ETDEWEB)

    Duran M, H. A.; Hernandez O, M. [Departamento de Investigacion en Polimeros y Materiales, Universidad de Sonora, Blvd. Luis Encinas y Rosales s/n, Col. Centro, 83190 Hermosillo, Sonora (Mexico); Salas L, M. A.; Hernandez D, V. M.; Vega C, H. R. [Unidad Academica de Estudios Nucleares, Universidad Autonoma de Zacatecas, Cipres 10, Fracc. La Penuela, 98068 Zacatecas (Mexico); Pinedo S, A.; Ventura M, J.; Chacon, F. [Hospital General de Zona No. 1, IMSS, Interior Alameda 45, 98000 Zacatecas (Mexico); Rivera M, T. [Centro de Investigacion en Ciencia Aplicada y Tecnologia Avanzada, IPN, Av. Legaria 694, Col. Irrigacion, 11500 Mexico D. F.(Mexico)], e-mail:


    Mammography test is a central tool for breast cancer diagnostic. In addition, programs are conducted periodically to detect the asymptomatic women in certain age groups; these programs have shown a reduction on breast cancer mortality. Early detection of breast cancer is achieved through a mammography, which contrasts the glandular and adipose tissue with a probable calcification. The parameters used for mammography are based on the thickness and density of the breast, their values depend on the voltage, current, focal spot and anode-filter combination. To achieve an image clear and a minimum dose must be chosen appropriate irradiation conditions. Risk associated with mammography should not be ignored. This study was performed in the General Hospital No. 1 IMSS in Zacatecas. Was used a glucose phantom and measured air Kerma at the entrance of the breast that was calculated using Monte Carlo methods and ZrO{sub 2}+PTFE thermoluminescent dosemeters, this calculation was completed with calculating the absorbed dose. (author)

  13. Use of a robotic manipulator in the simulation of the automation of a calibration process of dosemeters; Uso de un manipulador robotico en la simulacion de la automatizacion de un proceso de calibracion de dosimetros

    Energy Technology Data Exchange (ETDEWEB)

    Benitez R, J.S.; Najera H, M.C. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)


    The development of a system based in a manipulative robot which simulates the operative sequence in a calibration process of dosemeters is presented. In this process it is performed the monitoring of the dosemeter positions and the calibrator by mean of an arm of articulated robot which develops the movement sequences and the taking a decision based on the information coming from the external sensors. (Author)

  14. Methodology for Mixed Field Inhalation Dosimetry in Monazite Areas using a Twin-Cup Dosemeter with Three Track Detectors

    Energy Technology Data Exchange (ETDEWEB)

    Mayya, Y.S.; Eappen, K.P.; Nambi, K.S.V


    A methodology is proposed for estimating the gas and potential alpha energy concentrations (PAECs), and inhalation dose rates due to a mixed field of radon, thoron and their progeny present in the indoor atmosphere, using a radon-thoron discriminating dosemeter having three solid state nuclear track detectors (SSNTDs). The gas concentrations present in the vicinity of the dosemeter are directly measured through SSNTDs exposed in the 'cup mode'. The bare track density rates are related to the gas and the progeny concentrations by incorporating ventilation rate dependent spatial profiles for thoron gas. The ventilation rate is extracted as a real positive root of the governing equation and from this, the PAECs as well as the inhalation dose rates are estimated through the use of standard formulae. Error estimates on the dose rates have been carried out. Analyses of the field data from a sample survey of 35 houses situated in monazite bearing high background radiation areas of Kerala indicate that the methodology works successfully in a majority of the cases. Results for 22 cases which showed less than 100% statistical error are presented. These show median values of about 4.8 Bq.m{sup -3} and 462 Bq.m{sup -3} for radon and room averaged thoron concentrations, 0.5 mWL and 129 mWL for radon progeny and thoron progeny PAECs, and 1.21 {mu}Sv.h{sup -1} for inhalation dose rate. Although a complete picture would emerge only at the end of the ongoing large scale survey, the sample survey already indicates that indoor thoron levels in the Kerala monazite area are the highest reported so far in the literature. (author)

  15. A fibre optic scintillator dosemeter for absorbed dose measurements of low-energy X-ray-emitting brachytherapy sources. (United States)

    Sliski, Alan; Soares, Christopher; Mitch, Michael G


    A newly developed dosemeter using a 0.5 mm diameter x 0.5 mm thick cylindrical plastic scintillator coupled to the end of a fibre optic cable is capable of measuring the absorbed dose rate in water around low-activity, low-energy X-ray emitters typically used in prostate brachytherapy. Recent tests of this dosemeter showed that it is possible to measure the dose rate as a function of distance in water from 2 to 30 mm of a (103)Pd source of air-kerma strength 3.4 U (1 U = 1 microGy m(2) h(-1)), or 97 MBq (2.6 mCi) apparent activity, with good signal-to-noise ratio. The signal-to-noise ratio is only dependent on the integration time and background subtraction. The detector volume is enclosed in optically opaque, nearly water-equivalent materials so that there is no polar response other than that due to the shape of the scintillator volume chosen, in this case cylindrical. The absorbed dose rate very close to commercial brachytherapy sources can be mapped in an automated water phantom, providing a 3-D dose distribution with sub-millimeter spatial resolution. The sensitive volume of the detector is 0.5 mm from the end of the optically opaque waterproof housing, enabling measurements at very close distances to sources. The sensitive detector electronics allow the measurement of very low dose rates, as exist at centimeter distances from these sources. The detector is also applicable to mapping dose distributions from more complex source geometries such as eye applicators for treating macular degeneration.

  16. The Progress of HSP22 Research%HSP22研究进展

    Institute of Scientific and Technical Information of China (English)

    陶希; 卢伟; 胡治平; 郑丽君; 宋涛


    HSP22是小热休克蛋白超家族成员之一,主要在脑、心肌和骨骼肌广泛表达,而在子宫、前列腺、肺和肾脏等组织表达一般.当各种物理、化学等因素刺激时,可启动HSP22 mRNA快速表达;再通过HSP22 N末端区域和(或)C末端区域特定位点的突变或磷酸化反应,不同构象间相互转化及作用,在保持伴侣活性、激酶活性、触发炎性介质释放、抗凋亡与致凋亡和保护细胞骨架等方面起主要作用.其活性下降会导致遗传性末梢运动神经病、Alzheimer病、结蛋白相关心肌病、乳腺癌及白内障等相关疾病的发生发展.由于其在生命领域的特殊功能作用,目前生物医学界对其研究广泛.本文收集近期国内外文献,从分子和细胞水平综述了HSP22在基因表达、分子结构、生物学活性及与临床疾病关系的最新研究进展,以供学习交流.%HSP22 is a member of the superfamily of small heat-shock proteins, expressing predominantly in brain, heart and skeletal muscle, commonly in uterus , prostate , lung and kidney tissues. When activated by physical or chemistry factors , HSP22 mRNA express quickly in different tissues. Mutation in definite point or phosphorylation reaction in N terminal region and(or) C terminal region of HSP22, following reciprocal transformation and reaction in different conformation, which plays a critical role in the maintenance of chaperone activity, kinase activity, triggering the discharge of mediators of inflammation, anti-apoptosis or pro-apoptosis , protection of cytoskeleton, and so on. A decrease in the HSP22 activity may contribute to the development of heritage distal motor neuropathy, Alzheimer's disease, Desmin relevant cardiomyopathy, Breast cancer, Cataract, and so on. Because of its special functions and roles in life area, it was widely researched by persons in biomedicine territory now. In this review, we will collect literature of home and abroad and discuss recent

  17. Interactions of p60, a mediator of progesterone receptor assembly, with heat shock proteins hsp90 and hsp70

    DEFF Research Database (Denmark)

    Chen, S; Prapapanich, V; Rimerman, R A;


    mature PR complexes. In the present study we observe that a monoclonal antibody specific for p60 can, on the one hand, inhibit formation of mature PR complexes containing heat shock protein 90 (hsp90), p23, and immunophilins and, on the other, enhance recovery of early PR complexes containing hsp70...

  18. Hsp90 C-terminal inhibitors exhibit antimigratory activity by disrupting the Hsp90α/Aha1 complex in PC3-MM2 cells. (United States)

    Ghosh, Suman; Shinogle, Heather E; Garg, Gaurav; Vielhauer, George A; Holzbeierlein, Jeffrey M; Dobrowsky, Rick T; Blagg, Brian S J


    Human Hsp90 isoforms are molecular chaperones that are often up-regulated in malignances and represent a primary target for Hsp90 inhibitors undergoing clinical evaluation. Hsp90α is a stress-inducible isoform of Hsp90 that plays a significant role in apoptosis and metastasis. Though Hsp90α is secreted into the extracellular space under metastatic conditions, its role in cancer biology is poorly understood. We report that Hsp90α associates with the Aha1 co-chaperone and found this complex to localize in secretory vesicles and at the leading edge of migrating cells. Knockdown of Hsp90α resulted in a defect in cell migration. The functional role of Hsp90α/Aha1 was studied by treating the cells with various novobiocin-based Hsp90 C-terminal inhibitors. These inhibitors disrupted the Hsp90α/Aha1 complex, caused a cytoplasmic redistribution of Hsp90α and Aha1, and decreased cell migration. Structure-function studies determined that disruption of Hsp90α/Aha1 association and inhibition of cell migration correlated with the presence of a benzamide side chain, since an acetamide substituted analog was less effective. Our results show that disruption of Hsp90α/Aha1 interactions with novobiocin-based Hsp90 C-terminal inhibitors may limit the metastatic potential of tumors.

  19. Quercetin对晶状体上皮细胞HSP70、HSP27表达的调节作用%HSP70 and HSP27 expression in lens epithelial cells modulated by Quercetin

    Institute of Scientific and Technical Information of China (English)

    饶惠英; 姚克; 汤霞靖; 徐雯


    目的研究大鼠眼钝挫伤后晶状体上皮细胞(LECs)热休克蛋白(HSP)70、HSP27的表达,并给予喂饲Quercetin(HSP阻滞剂),观察Quercetin对LECs HSP70及HSP27表达的调节.方法SD大鼠48只,随机分成拍打组和Quercetin组,每组各24只24眼,右眼为实验眼.拍打组:20 g铁球20 cm高度拍打眼球100次.Quercetin组:给大鼠喂饲Quercetin(100 mg/kg),2~3 h后再拍打眼球.RT-PCR检测LECs HSP70、HSP27基因表达.结果钝挫性眼外伤可造成LECs HSP70基因表达的增强,拍打眼球后1 h HSP70表达开始升高,3 h后达到高峰,24 h后降至正常.Quercetin组HSP70基因表达随时间亦出现相应的提高,但与拍打组相比其峰值下降,差异有非常显著性意义.两组HSP27基因表达均无明显改变.结论钝挫性眼外伤中LECsHSP70表达的增强提示HSP70可能在钝挫性外伤性白内障形成过程中对晶状体变性蛋白起保护作用,预先喂饲Quercetin可抑制LECsHSP70基因的表达,其作用机制可能发生于HSP转录水平.

  20. Imaging of Hsp70-positive tumors with cmHsp70.1 antibody-conjugated gold nanoparticles

    Directory of Open Access Journals (Sweden)

    Gehrmann MK


    Full Text Available Mathias K Gehrmann,1 Melanie A Kimm,2 Stefan Stangl,1 Thomas E Schmid,1 Peter B Noël,2 Ernst J Rummeny,2 Gabriele Multhoff11Department of Radiation Oncology, 2Department of Diagnostic and Interventional Radiology, Klinikum rechts der Isar, Technische Universität München, Munich, GermanyAbstract: Real-time imaging of small tumors is still one of the challenges in cancer diagnosis, prognosis, and monitoring of clinical outcome. Targeting novel biomarkers that are selectively expressed on a large variety of different tumors but not normal cells has the potential to improve the imaging capacity of existing methods such as computed tomography. Herein, we present a novel technique using cmHsp70.1 monoclonal antibody-conjugated spherical gold nanoparticles for quantification of the targeted uptake of gold nanoparticles into membrane Hsp70-positive tumor cells. Upon binding, cmHsp70.1-conjugated gold nanoparticles but not nanoparticles coupled to an isotype-matched IgG1 antibody or empty nanoparticles are rapidly taken up by highly malignant Hsp70 membrane-positive mouse tumor cells. After 24 hours, the cmHsp70.1-conjugated gold nanoparticles are found to be enriched in the perinuclear region. Specificity for membrane Hsp70 was shown by using an Hsp70 knockout tumor cell system. Toxic side effects of the cmHsp70.1-conjugated nanoparticles are not observed at a concentration of 1–10 µg/mL. Experiments are ongoing to evaluate whether cmHsp70.1 antibody-conjugated gold nanoparticles are suitable for the detection of membrane-Hsp70-positive tumors in vivo.Keywords: heat shock protein 70, tumor biomarker, theranostics, multimodal CT, multispectral CT, k-edge

  1. Structure, Function and Regulation of the Hsp90 Machinery

    Directory of Open Access Journals (Sweden)

    Jing Li


    Full Text Available Heat shock protein 90 (Hsp90 is an ATP-dependent molecular chaperone which is essential in eukaryotes. It is required for the activation and stabilization of a wide variety of client proteins and many of them are involved in important cellular pathways. Since Hsp90 affects numerous physiological processes such as signal transduction, intracellular transport, and protein degradation, it became an interesting target for cancer therapy. Structurally, Hsp90 is a flexible dimeric protein composed of three different domains which adopt structurally distinct conformations. ATP binding triggers directionality in these conformational changes and leads to a more compact state. To achieve its function, Hsp90 works together with a large group of cofactors, termed co-chaperones. Co-chaperones form defined binary or ternary complexes with Hsp90, which facilitate the maturation of client proteins. In addition, posttranslational modifications of Hsp90, such as phosphorylation and acetylation, provide another level of regulation. They influence the conformational cycle, co-chaperone interaction, and inter-domain communications. In this review, we discuss the recent progress made in understanding the Hsp90 machinery.

  2. Metazoan Hsp70-based protein disaggregases: Emergence and mechanisms

    Directory of Open Access Journals (Sweden)

    Nadinath Bandara Nillegoda


    Full Text Available Proteotoxic stresses and ageing cause breakdown of cellular protein homeostasis, allowing misfolded proteins to form aggregates, which dedicated molecular machines have evolved to solubilize. In bacteria, fungi, protozoa and plants protein disaggregation involves an Hsp70-J-protein chaperone system, which loads and activates a powerful AAA+ ATPase (Hsp100 disaggregase onto protein aggregate substrates. Metazoans lack cytosolic and nuclear Hsp100 disaggregases but still eliminate protein aggregates. This longstanding puzzle of protein quality control is now resolved. Robust protein disaggregation activity recently shown for the metazoan Hsp70-based disaggregases relies instead on a crucial cooperation between two J-protein classes and interaction with the Hsp110 co-chaperone. An expanding multiplicity of Hsp70 and J-protein family members in metazoan cells facilitates different configurations of this Hsp70-based disaggregase allowing unprecedented versatility and specificity in protein disaggregation. Here we review the architecture, operation and adaptability of the emerging metazoan disaggregation system and discuss how this evolved.

  3. Structure and expression of a rice hsp70 gene

    Institute of Scientific and Technical Information of China (English)

    王群; 方荣祥


    A genomic hsp70 gene was isolated from a rice IR36 genomic library and 4 794 bp of the gene have been sequenoed. The 5’ flanking region of the gene contained a putative TATA box and a typical heat shock element sequence 5’-CTcgGAAccTTCgAG-3’. The amino acid sequence of the rice HSP70 deduced from the coding region shared 84%-92% homologies with those of HSP70s from other plant species. An intron 1939bp long was identified in the coding region at the codon specifying amino acid 72 (Asp), the similar position introns occurring in other intron-containing hsp70 genes. In addition, another intron of 57 bp was found in the 3’-untranslated region in the rice hsp70 gene. Southern blot hybridization showed that rice hsp70 gene family contained at least three members. Analysis of the RNA leveis with the gene-specific and non-specific probes revealed that the rice hsp70 gene expressed at normal temperature and the expression was enhanced by heat shock treatment.

  4. Hsp90 selectively modulates phenotype in vertebrate development.

    Directory of Open Access Journals (Sweden)

    Patricia L Yeyati


    Full Text Available Compromised heat shock protein 90 (Hsp90 function reveals cryptic phenotypes in flies and plants. These observations were interpreted to suggest that this molecular stress-response chaperone has a capacity to buffer underlying genetic variation. Conversely, the protective role of Hsp90 could account for the variable penetrance or severity of some heritable developmental malformations in vertebrates. Using zebrafish as a model, we defined Hsp90 inhibitor levels that did not induce a heat shock response or perturb phenotype in wild-type strains. Under these conditions the severity of the recessive eye phenotype in sunrise, caused by a pax6b mutation, was increased, while in dreumes, caused by a sufu mutation, it was decreased. In another strain, a previously unobserved spectrum of severe structural eye malformations, reminiscent of anophthalmia, microphthalmia, and nanophthalmia complex in humans, was uncovered by this limited inhibition of Hsp90 function. Inbreeding of offspring from selected unaffected carrier parents led to significantly elevated malformation frequencies and revealed the oligogenic nature of this phenotype. Unlike in Drosophila, Hsp90 inhibition can decrease developmental stability in zebrafish, as indicated by increased asymmetric presentation of anophthalmia, microphthalmia, and nanophthalmia and sunrise phenotypes. Analysis of the sunrise pax6b mutation suggests a molecular mechanism for the buffering of mutations by Hsp90. The zebrafish studies imply that mild perturbation of Hsp90 function at critical developmental stages may underpin the variable penetrance and expressivity of many developmental anomalies where the interaction between genotype and environment plays a major role.

  5. Overproduction and biophysical characterization of human HSP70 proteins. (United States)

    Boswell-Casteel, Rebba C; Johnson, Jennifer M; Duggan, Kelli D; Tsutsui, Yuko; Hays, Franklin A


    Heat shock proteins (HSP) perform vital cellular functions and modulate cell response pathways to physical and chemical stressors. A key feature of HSP function is the ability to interact with a broad array of protein binding partners as a means to potentiate downstream response pathways or facilitate protein folding. These binding interactions are driven by ATP-dependent conformational rearrangements in HSP proteins. The HSP70 family is evolutionarily conserved and is associated with diabetes and cancer progression and the etiopathogenesis of hepatic, cardiovascular, and neurological disorders in humans. However, functional characterization of human HSP70s has been stymied by difficulties in obtaining large quantities of purified protein. Studies of purified human HSP70 proteins are essential for downstream investigations of protein-protein interactions and in the rational design of novel family-specific therapeutics. Within this work, we present optimized protocols for the heterologous overexpression and purification of either the nucleotide binding domain (NBD) or the nucleotide and substrate binding domains of human HSPA9, HSPA8, and HSPA5 in either Escherichia coli or Saccharomyces cerevisiae. We also include initial biophysical characterization of HSPA9 and HSPA8. This work provides the basis for future biochemical studies of human HSP70 protein function and structure.

  6. Evaluation of neutron doses received at different organs in radiotherapy treatments using the UAB PADC based dosemeters in an anthropomorphic phantom

    Energy Technology Data Exchange (ETDEWEB)

    Domingo, C., E-mail: [Grup de Fisica de les Radiacions, Departament de Fisica, Edifici C, Universitat Autonoma de Barcelona, E-08193 Bellaterra (Spain); Garcia-Fuste, M.J.; Morales, E.; Amgarou, K.; Castelo, J. [Grup de Fisica de les Radiacions, Departament de Fisica, Edifici C, Universitat Autonoma de Barcelona, E-08193 Bellaterra (Spain); Sanchez-Doblado, F. [Hospital Universitario Virgen Macarena, Servicio de Radiofisica, Sevilla (Spain); Departamento de Fisiologia Medica y Biofisica, Facultad de Medicina, Universidad de Sevilla (Spain)


    The NEUTOR project was set up to study radiotherapy patient exposures to the neutrons produced around the LINAC accelerator head by photon radiation above approx8 MeV. These neutrons may reach the patient directly, or they may interact with the surrounding materials until they become thermalised, scattering all over the treatment room and affecting the patient as well. A miniaturised version of the UAB PADC based neutron dosemeter is used, together with thermoluminescence Li-6/Li-7 pairs, in several positions inside an anthropomorphic female phantom to determine doses received by the patient at several organs or tissues. Irradiations were made at a 15 MV LINAC (Hospital Universitario Virgen de la Macarena, Sevilla, Spain), an 18 MV LINAC (Hospital General Universitario de Valencia, Spain) and a 23 MV LINAC (Universitaetsklinikum Heidelberg, Germany). Preliminary patient dose maps obtained from the UAB PADC dosemeter measurements (considering an average calibration factor for all neutron energies) are reported in this work.

  7. Role of HSP100 proteins in plant stress tolerance. Final technical report

    Energy Technology Data Exchange (ETDEWEB)

    Vierling, E.


    This research focused on the following areas: characterization of HSP100 genes and their expression during stress and development; requirement of HSP101 for thermotolerance; thermotolerance of plants over-expressing HSP100; and identifying interacting proteins that functionally interact with HSP104.

  8. Bio Engineering Laboratory (United States)

    Federal Laboratory Consortium — Description/History: Chemistry and biology laboratoriesThe Bio Engineering Laboratory (BeL) is theonly full spectrum biotechnology capability within the Department...

  9. Bio Engineering Laboratory (United States)

    Federal Laboratory Consortium — Description/History: Chemistry and biology laboratories The Bio Engineering Laboratory (BeL) is theonly full spectrum biotechnology capability within the Department...

  10. Determination of the dose index in computerized tomography using thermoluminescent dosemeters; Determinacion del indice de dosis en tomografia computada usando dosimetros termoluminiscentes

    Energy Technology Data Exchange (ETDEWEB)

    Azorin, J.C.; Calderon, A. [Centro de Investigacion en Ciencia Aplicada y Tecnologia Avanzada-IPN, 11500 Mexico D.F. (Mexico); Azorin, J. [UAM-I, 09340 Mexico D.F. (Mexico)


    In this work the obtained results of the determination of the dose index are presented in thorax studies in computed tomography and helical tomography carried out in Mexico using thermoluminescent dosemeters of LiF: Mg,Cu,P + Ptfe developed and manufactured in our country. The results showed that under similar conditions of irradiation and operation (pitch = 1), significant differences don't exist among the doses absorbed measures in the phantom due to the two types of used tomographs. (Author)

  11. Conserved structure and expression of hsp70 paralogs in teleost fishes

    DEFF Research Database (Denmark)

    Metzger, David C.H.; Hansen, Jakob Hemmer; Schulte, Patricia M.


    The cytosolic 70 KDa heat shock proteins (Hsp70s) are widely used as biomarkers of environmental stress in ecological and toxicological studies in fish. Here we analyze teleost genome sequences to show that two genes encoding inducible hsp70s (hsp70-1 and hsp70-2) are likely present in all teleost...... fish. Phylogenetic and synteny analyses indicate that hsp70-1 and hsp70-2 are distinct paralogs that originated prior to the diversification of the teleosts. The promoters of both genes contain a TATA box and conserved heat shock elements (HSEs), but unlike mammalian HSP70s, both genes contain...

  12. Inducible HSP70 Protects Radiation-Induced Salivary Gland Damage

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hae-June; Lee, Yoon-Jin; Kwon, Hee-Choong; Lee, Su-Jae; Bae, Sang-Woo; Lee, Yun-Sil [Korea Institute of Radiological Medical Sciences, Seoul (Korea, Republic of); Kim, Sung-Ho [Chonnam National University, Gwangju (Korea, Republic of)


    Irradiation (IR) delivered to the head and neck is a common treatment for malignancies. Salivary glands in the irradiation field are severely damaged, and consequently this resulted in marked salivary hypofunction. While the exact mechanism of salivary gland damage remains enigmatic, fluid secreting acinar cells are lost, and saliva output is dramatically reduced. Previously we have reported that inducible heat shock protein 70 (HSP70i) induced radioresistance in vitro. Moreover, HSP70i localized to salivary glands by gene transfer has great potential for the treatment of salivary gland. Herein, we investigated whether HSP70 can use as radio protective molecules for radiation-induced salivary gland damage in vivo.

  13. HSP20 phosphorylation and airway smooth muscle relaxation

    Directory of Open Access Journals (Sweden)

    Mariam Ba


    Full Text Available Mariam Ba1, Cherie A Singer1, Manoj Tyagi2, Colleen Brophy3, Josh E Baker4, Christine Cremo4, Andrew Halayko5, William T Gerthoffer21Department of Pharmacology, University of Nevada School of Medicine, Reno, NV, USA; 2Department of Biochemistry and Molecular Biology, University of South Alabama, Mobile, AL, USA; 3Harrington Department of Biochemistry, Arizona State University, Tempe, AZ, USA; 4Department of Biochemistry and Molecular Biology, University of Nevada, Reno, NV, USA; 5Departments of Physiology and Internal Medicine, University of Manitoba, Winnipeg, MB, CanadaAbstract: HSP20 (HSPB6 is a small heat shock protein expressed in smooth muscles that is hypothesized to inhibit contraction when phosphorylated by cAMP-dependent protein kinase. To investigate this hypothesis in airway smooth muscle (ASM we showed that HSP20 was constitutively expressed as well as being inducible in cultured hASM cells by treatment with 1 µM isoproterenol or 10 µM salmeterol. In contrast, a mixture of proinflammatory mediators (interleukin-1β, tumor necrosis factor α, and interferon γ inhibited expression of HSP20 by about 50% in 48 hours. To determine whether phosphorylation of HSP20 is sufficient to induce relaxation, canine tracheal smooth muscle was treated with a cell permeant phosphopeptide that mimics the phosphorylation of HSP20. The HSP20 phosphopeptide antagonized carbacholinduced contraction by 60% with no change in myosin light chain phosphorylation. Recombinant full length HSP20 inhibited skeletal actin binding to smooth muscle myosin subfragment 1 (S1, and recombinant cell permeant TAT-HSP20 S16D mutant reduced F-actin filaments in cultured hASM cells. Carbachol stimulation of canine tracheal smooth muscle tissue caused redistribution of HSP20 from large macromolecular complexes (200–500 kDa to smaller complexes (<60 kDa. The results are consistent with HSP20 expression and macromolecular structure being dynamically regulated in airway

  14. The ribosomal biogenesis protein Utp21 interacts with Hsp90 and has differing requirements for Hsp90-associated proteins.

    Directory of Open Access Journals (Sweden)

    Victoria R Tenge

    Full Text Available The molecular chaperone Hsp90 buffers the effects of genetic variation by assisting the stabilization and folding of multiple clients critical for cell signaling and growth. We identified an interaction of Hsp90 and associated proteins with the essential nucleolar protein, Utp21, part of a large complex required for biogenesis of the small ribosomal subunit. The utp21-S602F mutation, which causes minor defects in otherwise wild-type yeast, exhibited severe or lethal growth defects when combined with mutations in Hsp90 or co-chaperones. WT Utp21 and Utp21-S602F exhibited similar interactions with Hsp90, and steady-state levels of WT Utp21 were reduced upon Hsp90 mutation or inhibition. Mutations in the human homolog of UTP21, WDR36, have been associated with adult-onset primary open-angle glaucoma, a leading cause of blindness worldwide. Three different mutant forms of Utp21 analogous to glaucoma-associated WDR36 mutations exhibit reduced levels in yeast cells expressing mutations in Hsp90 or associated chaperones, suggesting that Hsp90 and co-chaperones buffer the effects of those mutations.

  15. Effect of methylglyoxal modification on the structure and properties of human small heat shock protein HspB6 (Hsp20). (United States)

    Muranova, Lydia K; Perfilov, Maxim M; Serebryakova, Marina V; Gusev, Nikolai B


    Human small heat shock protein HspB6 (Hsp20) was modified by metabolic α-dicarbonyl compound methylglyoxal (MGO). At low MGO/HspB6 molar ratio, Arg13, Arg14, Arg27, and Arg102 were the primary sites of MGO modification. At high MGO/HspB6 ratio, practically, all Arg and Lys residues of HspB6 were modified. Both mild and extensive MGO modification decreased susceptibility of HspB6 to trypsinolysis and prevented its heat-induced aggregation. Modification by MGO was accompanied by formation of small quantities of chemically crosslinked dimers and did not dramatically affect quaternary structure of HspB6. Mild modification by MGO did not affect whereas extensive modification decreased interaction of HspB6 with HspB1. Phosphorylation of HspB6 by cyclic adenosine monophosphate (cAMP)-dependent protein kinase was inhibited after mild modification and completely prevented after extensive modification by MGO. Chaperone-like activity of HspB6 measured with subfragment 1 of skeletal myosin was enhanced after MGO modifications. It is concluded that Arg residues located in the N-terminal domain of HspB6 are easily accessible to MGO modification and that even mild modification by MGO affects susceptibility to trypsinolysis, phosphorylation by cAMP-dependent protein kinase, and chaperone-like activity of HspB6.

  16. Phage displaying epitope of Candida albicans HSP90 and serodiagnosis

    Institute of Scientific and Technical Information of China (English)

    杨琼; 王丽; 卢大宁; 邢沈阳; 尹东; 朱筱娟


    @@ Recently, the frequent use of immunosuppressants and chemotherapeutic drugs for cancers has caused an increase in the frequency of life-threatening systemic candidiasis.1 Studies by Matthews et al2 indicated HSP90 fragments are major targets for the immune system in infection due to C. albicans, and anti-epitope LKVIRK of HSP90 antibody is a serological marker for diagnosis of invasive candidiasis. Cloning and sequencing HSP90 antigen revealed that the linear epitope LKVIRK, localized near the C-terminus of the 47 kDa protein which circulates in the sera of patients with invasive candidiasis, as a heat-stable breakdown product of large more heat-labile antigen HSP90.2 In this study, epitope LKVIRK was displayed on the surface of phage fd to develop a new serological test for systemic candidiasis.

  17. A role for anti-HSP60 antibodies in arthritis

    DEFF Research Database (Denmark)

    Carlsen, Thomas Gelsing; Bennike, Tue; Christiansen, Gunna;


    As a result of the high sequence similarity between HSP60 proteins, found in both prokaryotic and eukaryotic cells, it has been suggested, but never concluded, that anti-HSP60 antibodies could be of importance in the pathology of arthritis diseases explained by a concept named molecular mimicry......60. In this review, these new findings are compared with old questioning the durability of molecular mimicry as a hypothesis for arthritis pathogenesis....

  18. Perbedaan Kadar HSP90 pada Preeklamsi Berat dengan Kehamilan Normal

    Directory of Open Access Journals (Sweden)



    Full Text Available Severe pre-eclampsia is the second highest cause of maternal mortality. Free radicals that stimulate heat shock protein 90 (HSP 90 are believed to determine severe pre-eclampsia. HSP90 is an important protein that helps the establishment and maintenance of other proteins. It also increases the life time of cells after various pathological conditions (chaperone function. The chaperone function is the adaptation key factor to endogenous stress in tissues. By recognizing HSP90 level in early detection of severe pre-eclampsia, prevention and management can be started early. This study aimed to prove that the HSP90 level in pregnancy with severe pre-eclampsia is higher than normal pregnancy. This was a quantitative study using cross sectional approach by testing the HSP90 level. The study was conducted during the period of September to November 2013, at the Obstetrics and Gynecological Unit, Moewardi Hospital Surakarta and Prodia Laboratory Jakarta. The number of subjects was 30 patients, consisting of 15 normal pregnant mothers and 15 pregnant mothers with pre-eclampsia . The calculation of serum HSP90 level was conducted using enzyme-linked immunosorbent assay (ELISA. Data were analyzed using t-test using SPSS for Windows version 17 for Windows. The mean of HSP90 in the severe pre-eclampsia group was 131.91±26.66 while the mean in the normal pregnancy group was 80.28±13.39 with p=0.00 (p<0.05. Level of HSP90 serum in severe pre-eclampsia is higher than in normal pregnancy, due to the occurrence of oxidative stress in severe pre-eclampsia

  19. Research Progress of HSP90 in Aquatic Animals and Plants%HSP90及其在水生动植物中的研究进展

    Institute of Scientific and Technical Information of China (English)



    The classification, structure and transcription regulation of HSP90s were briefly reviewed. Special attention was paid to inducing expression and physiological function of HSP90s in aquatic animals and plants.%简要介绍了HSP90的分类、结构和转录调控,重点讨论了HSP90在水产动植物中的诱导表达与生理功能.

  20. Contribution of IL-6 to the Hsp72, Hsp25, and alphaB-crystallin [corrected] responses to inflammation and exercise training in mouse skeletal and cardiac muscle. (United States)

    Huey, Kimberly A; Meador, Benjamin M


    The heat shock proteins (Hsps) Hsp72, Hsp25, and alphaB-crystallin (alphaB C) [corrected]may protect tissues during exercise and/or inflammatory insults; however, no studies have investigated whether exercise training increases both basal and inflammation-induced expression of these Hsps in skeletal or cardiac muscle. IL-6 is produced by muscle during both exercise and inflammation and has been shown to modulate Hsp expression. These studies tested the hypothesis that voluntary wheel running (RW) increases basal and inflammation-induced Hsp72, Hsp25, and alphaB C [corrected] protein through an IL-6-dependent mechanism. We compared Hsp72, Hsp25, alphaB C, [corrected] and IL-6 protein levels 4 h after systemic inflammation induced by lipopolysaccharide (LPS) in skeletal and cardiac muscles of wild-type (IL-6(+/+)) and IL-6 deficient (IL-6(-/-)) mice after 2 wk of RW or normal cage activity (Sed). LPS significantly increased skeletal Hsp72 and Hsp25 relative to saline in Sed IL-6(+/+), but not IL-6(-/-) mice. LPS increased Hsp72 relative to saline in Sed IL-6(+/+) cardiac muscle. RW increased basal Hsp72, Hsp25, and alphaB C [corrected] in skeletal muscle in IL-6(+/+) and IL-6(-/-) mice. However, LPS was not associated with increases in any Hsp in RW IL-6(+/+) or IL-6(-/-) mice. LPS increased IL-6 protein in skeletal muscle and plasma in Sed and RW groups, with a significantly greater response in RW. The major results provide the first in vivo evidence that the absence of IL-6 is associated with reduced skeletal muscle Hsp72 and Hsp25 responses to LPS, but that IL-6 is not required for exercise-induced Hsp upregulation in skeletal or cardiac muscle.

  1. Application of Bioinformatics in the Study of HSP70 Recombinant Vaccine of Echinococcus Granulosus%生物信息学技术在细粒棘球蚴热休克蛋白70重组疫苗研究中的应用

    Institute of Scientific and Technical Information of China (English)

    丁淑琴; 刘宏鹏; 张爱君; 张彩芳; 赵巍


    目的 通过基因序列分析,寻找细粒棘球蚴(Echinococcus granulosus,Eg)热休克蛋白基因(Heat shock pm-tein7,HSP70),为包虫病防治筛选新的候选疫苗抗原.方法 ①登录 GenBank 公共数据库,检索 EgHSP70目的基因序列.②对克隆基因进行测序,结果输入DNAStax、BioSun分析软件和互联网 SWISS-MODEI 数据库,对重组 EgHSP70 的特性、抗原表位及构象进行预测分析.结果 ①获得了具有完整开放阅读框的 EgHSP70核苷酸序列.②DNAStar分析软件推算氨基酸序列结果显示,EgHSP70由133个理论氨基酸组成,分子量约为14.53kDa.DNAStar、BioSun分析软件确定了重组 EgHSP70 可能的抗原表位.与其他动物热休克蛋白70氨基酸的同源性分析结果显示EgHSP70具有高度的保守性.结论 生物信息学技术在 EgHSP70 重组疫苗研究中有一定的理论和应用价值.

  2. Hsp100/ClpB Chaperone Function and Mechanism

    Energy Technology Data Exchange (ETDEWEB)

    Vierling, Elizabeth [Univ. of Massachusetts, Amherst, MA (United States). Dept. of Biochemistry and Molecular Biology


    The supported research investigated the mechanism of action of a unique class of molecular chaperones in higher plants, the Hsp100/ClpB proteins, with the ultimate goal of defining how these chaperones influence plant growth, development, stress tolerance and productivity. Molecular chaperones are essential effectors of cellular “protein quality control”, which comprises processes that ensure the proper folding, localization, activation and turnover of proteins. Hsp100/ClpB proteins are required for temperature acclimation in plants, optimal seed yield, and proper chloroplast development. The model plant Arabidopsis thaliana and genetic and molecular approaches were used to investigate two of the three members of the Hsp100/ClpB proteins in plants, cytosolic AtHsp101 and chloroplast-localized AtClpB-p. Investigating the chaperone activity of the Hsp100/ClpB proteins addresses DOE goals in that this activity impacts how “plants generate and assemble components” as well as “allowing for their self repair”. Additionally, Hsp100/ClpB protein function in plants is directly required for optimal “utilization of biological energy” and is involved in “mechanisms that control the architecture of energy transduction systems”.

  3. Hsp60 and human aging: Les liaisons dangereuses. (United States)

    Cappello, Francesco; Conway de Macario, Everly; Marino Gammazza, Antonella; Bonaventura, Giuseppe; Carini, Francesco; Czarnecka, Anna M; Farina, Felicia; Zummo, Giovanni; Macario, Alberto J L


    Stressors can cause abnormal intracellular accumulation of Hsp60 and its localization in extramitochondrial sites, secretion, and circulation, with immune system activation. Dysfunction of chaperones associated with their quantitative and qualitative decline with aging (chaperonopathies of aging) characterizes senescence and is a potential causal factor in the physiological deterioration that occurs with it. The role of Hsp60 in aging is not easy to elucidate, because aging is accompanied by pathologies (e.g., cardiovascular and neurodegenerative disorders, osteoporosis, diabetes, cancer, etc.) in which Hsp60 has been implicated but, although those disorders are more frequent in the elderly, they are not unique to them. Therefore, it is difficult to determine what is due to aging and what to an associated disease that can occur regardless of age. Does Hsp60 contribute to the pathogenesis? How and when does Hsp60 interact with the immune system and, thus, contributes to the initiation-progression of the generalized chronic inflammation characteristic of aging? These and related issues are discussed here in the light of reports showing the participation of Hsp60 in aging-associated disorders.

  4. Hsp10: anatomic distribution, functions, and involvement in human disease. (United States)

    David, Sabrina; Bucchieri, Fabio; Corrao, Simona; Czarnecka, Anna M; Campanella, Claudia; Farina, Felicia; Peri, Giovanni; Tomasello, Giovanni; Sciumè, Carmelo; Modica, Giuseppe; La Rocca, Giampiero; Anzalone, Rita; Giuffrè, Mario; Conway De Macario, Everly; Macario, Alberto J L; Cappello, Francesco; Zummo, Giovanni


    There is growing evidence that molecular chaperones/heat shock proteins are involved in the pathogenesis of a number of human diseases, known as chaperonopathies. A better molecular understanding of the pathogenetic mechanisms is essential for addressing new strategies in diagnostics, therapeutics and clinical management of chaperonopathies, including those in which Hsp10 is involved. This chaperonin has been studied for a long time as a member of the mitochondrial protein-folding machine. However, although in normal cells Hsp10 is mainly localized in the mitochondrial matrix, it has also been found during and after stress in other subcellular compartments, such as cytosol, vesicles and secretory granules, alone or in combination with other proteins. In these extramitochondrial locales, Hsp10 plays an active role in cell signalling. For example, cancer cells often show altered levels of Hsp10, compared to normal cells. Hsp10 may also be found in the extracellular space and in the bloodstream, with a possible immunomodulatory activity. This minireview focuses on some studies to date on the involvement of Hsp10 in human disease pathogenesis.

  5. Hsp60 is actively secreted by human tumor cells.

    Directory of Open Access Journals (Sweden)

    Anna M Merendino

    Full Text Available BACKGROUND: Hsp60, a Group I mitochondrial chaperonin, is classically considered an intracellular chaperone with residence in the mitochondria; nonetheless, in the last few years it has been found extracellularly as well as in the cell membrane. Important questions remain pertaining to extracellular Hsp60 such as how generalized is its occurrence outside cells, what are its extracellular functions and the translocation mechanisms that transport the chaperone outside of the cell. These questions are particularly relevant for cancer biology since it is believed that extracellular chaperones, like Hsp70, may play an active role in tumor growth and dissemination. METHODOLOGY/PRINCIPAL FINDINGS: Since cancer cells may undergo necrosis and apoptosis, it could be possible that extracellular Hsps are chiefly the result of cell destruction but not the product of an active, physiological process. In this work, we studied three tumor cells lines and found that they all release Hsp60 into the culture media by an active mechanism independently of cell death. Biochemical analyses of one of the cell lines revealed that Hsp60 secretion was significantly reduced, by inhibitors of exosomes and lipid rafts. CONCLUSIONS/SIGNIFICANCE: Our data suggest that Hsp60 release is the result of an active secretion mechanism and, since extracellular release of the chaperone was demonstrated in all tumor cell lines investigated, our observations most likely reflect a general physiological phenomenon, occurring in many tumors.

  6. Heat Shock Protein 90 (Hsp90 Expression and Breast Cancer

    Directory of Open Access Journals (Sweden)

    Christos A. Papadimitriou


    Full Text Available Hsp90 is an abundant protein in mammalian cells. It forms several discrete complexes, each containing distinct groups of co-chaperones that assist protein folding and refolding during stress, protein transport and degradation. It interacts with a variety of proteins that play key roles in breast neoplasia including estrogen receptors, tumor suppressor p53 protein, angiogenesis transcription factor HIF-1alpha, antiapoptotic kinase Akt, Raf-1 MAP kinase and a variety of receptor tyrosine kinases of the erbB family. Elevated Hsp90 expression has been documented in breast ductal carcinomas contributing to the proliferative activity of breast cancer cells; whilst a significantly decreased Hsp90 expression has been shown in infiltrative lobular carcinomas and lobular neoplasia. Hsp90 overexpression has been proposed as a component of a mechanism through which breast cancer cells become resistant to various stress stimuli. Therefore, pharmacological inhibition of HSPs can provide therapeutic opportunities in the field of cancer treatment. 17-allylamino,17-demethoxygeldanamycin is the first Hsp90 inhibitor that has clinically been investigated in phase II trial, yielding promising results in patients with HER2-overexpressing metastatic breast cancer, whilst other Hsp90 inhibitors (retaspimycin HCL, NVP-AUY922, NVP-BEP800, CNF2024/BIIB021, SNX-5422, STA-9090, etc. are currently under evaluation.

  7. Developmental and hyperthermia-induced expression of the heat shock proteins HSP60 and HSP70 in tissues of the housefly Musca domestica: an in vitro study

    Directory of Open Access Journals (Sweden)

    Sunita Sharma


    Full Text Available The expression pattern of two major chaperones, the heat shock proteins (HSPs HSP60 and HSP70 was studied in vitro in tissues of the housefly Musca domestica during larval and adult stages of development to identify their immunological relatives and understand their functional significance in normal cellular activities and during thermal stress. Fluorographs of labeled polypeptides and western blots demonstrated that both HSPs are expressed constitutively and heat-induced in all the larval and adult cell types examined. The pattern of whole tissue immunocytochemical staining using anti-HSP60 and anti-HSP70 antibodies corresponded well with the observations from western blots or fluorographs. In developing oocytes, both constitutive and heat inducible expression of HSP60 were regulated in an oocyte stage-specific manner. In unstressed ovaries the expression of these proteins was less pronounced in early stage oocytes (1st - 8th than at later stages (9th and onward. The heat shock, however, induced both HSP70 and HSP60 to a significantly high level in early stage oocytes (1st-8th as compared to their respective controls. Our findings indicate the involvement of the HSP60 and HSP70 proteins in the development, growth and differentiation of both somatic and germ line tissues. Furthermore, the enhanced co-expression of HSP70 and HSP60 upon heat shock in various larval and adult cell types suggests the possible role of HSP60 in thermoprotection.

  8. Bios data analyzer. (United States)

    Sabelli, H; Sugerman, A; Kovacevic, L; Kauffman, L; Carlson-Sabelli, L; Patel, M; Konecki, J


    The Bios Data Analyzer (BDA) is a set of computer programs (CD-ROM, in Sabelli et al., Bios. A Study of Creation, 2005) for new time series analyses that detects and measures creative phenomena, namely diversification, novelty, complexes, nonrandom complexity. We define a process as creative when its time series displays these properties. They are found in heartbeat interval series, the exemplar of bios .just as turbulence is the exemplar of chaos, in many other empirical series (galactic distributions, meteorological, economic and physiological series), in biotic series generated mathematically by the bipolar feedback, and in stochastic noise, but not in chaotic attractors. Differencing, consecutive recurrence and partial autocorrelation indicate nonrandom causation, thereby distinguishing chaos and bios from random and random walk. Embedding plots distinguish causal creative processes (e.g. bios) that include both simple and complex components of variation from stochastic processes (e.g. Brownian noise) that include only complex components, and from chaotic processes that decay from order to randomness as the number of dimensions is increased. Varying bin and dimensionality show that entropy measures symmetry and variety, and that complexity is associated with asymmetry. Trigonometric transformations measure coexisting opposites in time series and demonstrate bipolar, partial, and uncorrelated opposites in empirical processes and bios, supporting the hypothesis that bios is generated by bipolar feedback, a concept which is at variance with standard concepts of polar and complementary opposites.

  9. Tl response of LiF: Mg, Cu, P + PTFE (Mexico) and GR200A (China) dosemeters; Respuesta Tl de dosimetros de LiF: Mg, Cu, P + PTFE (Mexico) y GR200A (China)

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez M, P.R. [ININ, A.P. 18-1027, Mexico D.F. (Mexico); Azorin N, J.; Furetta, C. [UAM-I, 09340 Mexico D.F. (Mexico)


    TLD-100 was the commercial dosemeter more known since some decades ago. This dosemeter was considered for many research groups as the reference material for developing new Tl materials for ionizing radiation dosimetry. Actually it seems that TLD-100 is going to be replaced by the Chinese material GR200A, as reference material due that this material in addition to be considered as a tissue equivalent material, is 30 to 35 times more sensitive that TLD- 100. Results of the study of the Tl response of LiF: Mg, Cu, P + Ptfe developed at ININ-Mexico, comparing them with those of GR200A are presented. These results showed that the sensitivity (s = Tl intensity/weight x dose) to gamma radiation of the LiF: Mg, Cu, P + Ptfe dosimeters was 4.34 meanwhile that of the commercial dosemeter was 3.41. Detection threshold of the dosemeters studied was 2.22 and 0.52 {mu}Gy respectively Repeatability after 10 cycles Irradiation-reading-annealing was {+-}1.39% and {+-}1.86% respectively. Both types of dosemeters presented a linear response as a function of gamma radiation in the range from 0.02 mGy and 100 Gy. (Author)

  10. Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself

    Directory of Open Access Journals (Sweden)

    Luong Michael


    Full Text Available Abstract Background Toll-like receptor 4 (TLR4 is activated by bacterial endotoxin, a prototypical pathogen-associated molecular pattern (PAMP. It has been suggested that TLR4 can also be activated by damage-associated molecular pattern (DAMP proteins such as HSP70. It remains a challenge to provide unequivocal evidence that DAMP proteins themselves play a role in TLR4 activation, as the DAMP proteins used are often contaminated with endotoxin and other TLR ligands introduced during protein expression and/or purification. Results Here we report that the activation of TLR4 on primary human macrophage cultures by recombinant HSP70 is not solely due to contaminating endotoxin. Polymyxin B pretreatment of HSP70 preparations to neutralize contaminating endotoxin caused significant reductions in the amount of TNF-α induced by the recombinant protein as determined by ELISA. However, digestion of HSP70 with Proteinase K-agarose beads also dramatically reduced the TNF-α response of macrophages to HSP70, while leaving levels of contaminating endotoxin largely unchanged relative to controls. Conclusions These results indicate that the stimulatory effect of recombinant HSP70 requires both the presence of endotoxin and structural integrity of the heat shock protein itself.

  11. Measurement of doses to aviator pilots using thermoluminescent dosemeters; Medicion de la dosis a pilotos aviadores usando dosimetros termoluminiscentes

    Energy Technology Data Exchange (ETDEWEB)

    Azorin N, J.; Cruz C, D. [UAM-I, 09340 Mexico D.F. (Mexico); Rivera M, T. [IPN, ESIME Culhuacan (Mexico)]. e-mail:


    During the development of their work, the aviator pilots are exposed at high levels of natural radiation of bottom caused mainly by the cosmic radiation of galactic origin and lot. For such reason, the Metropolitan Autonomous University (UAM) and the Union Association of Aviator Pilots (ASPA), subscribed an agreement with the purpose of to measure the doses of ionizing radiation received by the aviator pilots of diverse air companies that man different types of airships and to determine if these doses surpass the one limit of 0.11 mSv/h settled down by the IAEA for the public in general; and if therefore, these workers should be considered as personnel occupationally exposed. In this work the obtained results when measuring the absorbed dose received by Mexican civil aviator pilots during the development of their work, using thermoluminescent dosemeters of LiF:Mg,Cu,P + Ptfe of national production are presented. The obtained results during the years of 2001 and 2002 show that the monthly doses received by the pilots surpass the one it limits established for the public in general, for what they should be considered as personnel occupationally exposed. (Author)

  12. Induction of Hsp22 (HspB8) by estrogen and the metalloestrogen cadmium in estrogen receptor-positive breast cancer cells. (United States)

    Sun, Xiankui; Fontaine, Jean-Marc; Bartl, Ingrid; Behnam, Babak; Welsh, Michael J; Benndorf, Rainer


    Estrogen (E2) plays a critical role in the etiology and progression of human breast cancer. The estrogenic response is complex and not completely understood, including in terms of the involved responsive genes. Here we show that Hsp22 (synonyms: HspB8, E2lG1, H11), a member of the small heat shock protein (sHSP) superfamily, was induced by E2 in estrogen receptor-positive MCF-7 breast cancer cells, resulting in an elevated Hsp22 protein level, whereas it was not induced in estrogen receptor-negative MDA-MB-231 cells. This induction was prevented by the pure anti-estrogen ICI182780 (faslodex, fulvestrant), whereas tamoxifen, a substance with mixed estrogenic and antiestrogenic properties, had no major inhibitory effect on this induction, nor did it induce Hsp22 on its own. Cadmium (Cd) is an environmental pollutant with estrogenic properties (metalloestrogen) that has been implicated in breast cancer. Treatment of MCF-7 cells with Cd also resulted in induction of Hsp22, and this induction was also inhibited by ICI182780. In live MCF-7 cells, Hsp22 interacted at the level of dimers with Hsp27, a related sHSP, as was shown by quantitative fluorescence resonance energy transfer measurements. In cytosolic extracts of MCF-7 cells, most of the E2- and Cd-induced Hsp22 was incorporated into high-molecular mass complexes. In part, Hsp22 and Hsp27 were components of distinct populations of these complexes. Finally, candidate elements in the Hsp22 promoter were identified by sequence analysis that could account for the induction of Hsp22 by E2 and Cd. Taken together, Hsp22 induction represents a new aspect of the estrogenic response with potential significance for the biology of estrogen receptor-positive breast cancer cells.

  13. BioProject (United States)

    U.S. Department of Health & Human Services — The BioProject database provides an organizational framework to access information about research projects with links to data that have been or will be deposited...

  14. BioSentinel Project (United States)

    National Aeronautics and Space Administration — BioSentinel technology will provide critical information about how living systems, from humans down to cells, adapt, respond and survive in deep space, beyond LEO...

  15. BioSystems (United States)

    U.S. Department of Health & Human Services — The NCBI BioSystems Database provides integrated access to biological systems and their component genes, proteins, and small molecules, as well as literature...

  16. Engineering BioBrick vectors from BioBrick parts

    Directory of Open Access Journals (Sweden)

    Knight Thomas F


    Full Text Available Abstract Background The underlying goal of synthetic biology is to make the process of engineering biological systems easier. Recent work has focused on defining and developing standard biological parts. The technical standard that has gained the most traction in the synthetic biology community is the BioBrick standard for physical composition of genetic parts. Parts that conform to the BioBrick assembly standard are BioBrick standard biological parts. To date, over 2,000 BioBrick parts have been contributed to, and are available from, the Registry of Standard Biological Parts. Results Here we extended the same advantages of BioBrick standard biological parts to the plasmid-based vectors that are used to provide and propagate BioBrick parts. We developed a process for engineering BioBrick vectors from BioBrick parts. We designed a new set of BioBrick parts that encode many useful vector functions. We combined the new parts to make a BioBrick base vector that facilitates BioBrick vector construction. We demonstrated the utility of the process by constructing seven new BioBrick vectors. We also successfully used the resulting vectors to assemble and propagate other BioBrick standard biological parts. Conclusion We extended the principles of part reuse and standardization to BioBrick vectors. As a result, myriad new BioBrick vectors can be readily produced from all existing and newly designed BioBrick parts. We invite the synthetic biology community to (1 use the process to make and share new BioBrick vectors; (2 expand the current collection of BioBrick vector parts; and (3 characterize and improve the available collection of BioBrick vector parts.

  17. BioArchitecture


    Gunning, Peter


    Volume 1 has defined the scope of BioArchitecture. From the outset we have strived to ensure that BioArchitecture is not limited to the three major polymer systems of the cytoplasm. I am happy to say that a cursory glance at the contents of volume 1 makes it clear that we are interested in all aspects of bioarchitecture from molecules to polymers to cells to tissue to the organism.

  18. The former tests realized to a personal neutron dosemeter based on solid nuclear tracks detector; Primeras pruebas realizadas a un dosimetro personal de neutrones basado en detectores solidos de trazas nucleares

    Energy Technology Data Exchange (ETDEWEB)

    Camacho, M.E.; Tavera, L.; Balcazar, M. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)


    Due to the increase in the use of neutron radiation a personal neutron dosemeter based on solid nuclear tracks detector (DSTN) was designed and constructed. The personal dosemeter design consists of three arrangements. The first one consists of a plastic nuclear tracks detector (LR115 or CR39) in contact with a LiF pellet. The second one is the same that above but it placed among two cadmium pellets and, the third one is formed by the alone detector without converter neither neutron absorber. The three arrangements are placed inside a plastic porta detector hermetically closed to avoid the bottom produced by environmental radon whichever both detectors (LR115 and CR39) are sensitive. In this work the former tests realized to that dosemeter are presented. (Author)

  19. Study on the Preparation of Recombinant Human HSP70 and Its Presenting-Antigen Function

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Guimei(


    [1]Yasuaki T Norioki T Noriyuki S et al.70 kDa heat shock cognate protein is a transformation-associated antigen and a possible target for the host's anti-tumor immunity.J Immunol 1993 151:5516[2]Yasuaki T Ping P Kang L et al.Immunotherapy of tumors with autologous tumor-derived heat shock protein preparations.Science 1997 278:117[3]Heiichiro V Pramod K S.Heat shock protein 70-associated peptides elicit specific cancer immunity.J Exp Med 1998 178:1391[4]Sambrook J Fritch E F Maniatis T.Molecular Cloning a laboratory manu 2nd ed.New York:Cold Spring Harbor Laboratory Press 1989.253[5]Satish J Peter M Stanley R et al.Human stress protein hsp70:overexpression in E.coli purification and characterization.Bio/Technology 1995 113:1105[6]哈密斯B.D. 利克伍德D.著.刘毓秀 程桂芳译.蛋白质的凝胶电泳实践方法.北京:科学出版社 1994.151[7]Anne-marie T C Max P Carey L O et al.Immunization with a lymphocytic choriomeningitis virus peptide mixed with heat shock protein 70 results in protective antiviral immunity and specific cytotoxic T lymphocytes.J Exp Med 1998 187:685[8]Nandan D Daubenberger C Mpimbaza G.A rapid single-step purification method for immunogenic members of the Hsp70 family:validation and application.J Immunol Methods 1994 176(2) :255

  20. Tumor-intrinsic and tumor-extrinsic factors impacting hsp90- targeted therapy. (United States)

    Alarcon, S V; Mollapour, M; Lee, M-J; Tsutsumi, S; Lee, S; Kim, Y S; Prince, T; Apolo, A B; Giaccone, G; Xu, W; Neckers, L M; Trepel, J B


    In 1994 the first heat shock protein 90 (Hsp90) inhibitor was identified and Hsp90 was reported to be a target for anticancer therapeutics. In the past 18 years there have been 17 distinct Hsp90 inhibitors entered into clinical trial, and the small molecule Hsp90 inhibitors have been highly valuable as probes of the role of Hsp90 and its client proteins in cancer. Although no Hsp90 inhibitor has achieved regulatory approval, recently there has been significant progress in Hsp90 inhibitor clinical development, and in the past year RECIST responses have been documented in HER2-positive breast cancer and EML4-ALK-positive non-small cell lung cancer. All of the clinical Hsp90 inhibitors studied to date are specific in their target, i.e. they bind exclusively to Hsp90 and two related heat shock proteins. However, Hsp90 inhibitors are markedly pleiotropic, causing degradation of over 200 client proteins and impacting critical multiprotein complexes. Furthermore, it has only recently been appreciated that Hsp90 inhibitors can, paradoxically, cause transient activation of the protein kinase clients they are chaperoning, resulting in initiation of signal transduction and significant physiological events in both tumor and tumor microenvironment. An additional area of recent progress in Hsp90 research is in studies of the posttranslational modifications of Hsp90 itself and Hsp90 co-chaperone proteins. Together, a picture is emerging in which the impact of Hsp90 inhibitors is shaped by the tumor intracellular and extracellular milieu, and in which Hsp90 inhibitors impact tumor and host on a microenvironmental and systems level. Here we review the tumor intrinsic and extrinsic factors that impact the efficacy of small molecules engaging the Hsp90 chaperone machine.

  1. Role of Hsp104 in the propagation and inheritance of the [Het-s] prion. (United States)

    Malato, Laurent; Dos Reis, Suzana; Benkemoun, Laura; Sabaté, Raimon; Saupe, Sven J


    The chaperones of the ClpB/HSP100 family play a central role in thermotolerance in bacteria, plants, and fungi by ensuring solubilization of heat-induced protein aggregates. In addition in yeast, Hsp104 was found to be required for prion propagation. Herein, we analyze the role of Podospora anserina Hsp104 (PaHsp104) in the formation and propagation of the [Het-s] prion. We show that DeltaPaHsp104 strains propagate [Het-s], making [Het-s] the first native fungal prion to be propagated in the absence of Hsp104. Nevertheless, we found that [Het-s]-propagon numbers, propagation rate, and spontaneous emergence are reduced in a DeltaPaHsp104 background. In addition, inactivation of PaHsp104 leads to severe meiotic instability of [Het-s] and abolishes its meiotic drive activity. Finally, we show that DeltaPaHSP104 strains are less susceptible than wild type to infection by exogenous recombinant HET-s(218-289) prion amyloids. Like [URE3] and [PIN(+)] in yeast but unlike [PSI(+)], [Het-s] is not cured by constitutive PaHsp104 overexpression. The observed effects of PaHsp104 inactivation are consistent with the described role of Hsp104 in prion aggregate shearing in yeast. However, Hsp104-dependency appears less stringent in P. anserina than in yeast; presumably because in Podospora prion propagation occurs in a syncitium.

  2. A cytosolic relay of heat shock proteins HSP70-1A and HSP90β monitors the folding trajectory of the serotonin transporter. (United States)

    El-Kasaby, Ali; Koban, Florian; Sitte, Harald H; Freissmuth, Michael; Sucic, Sonja


    Mutations in the C terminus of the serotonin transporter (SERT) disrupt folding and export from the endoplasmic reticulum. Here we examined the hypothesis that a cytosolic heat shock protein relay was recruited to the C terminus to assist folding of SERT. This conjecture was verified by the following observations. (i) The proximal portion of the SERT C terminus conforms to a canonical binding site for DnaK/heat shock protein of 70 kDa (HSP70). A peptide covering this segment stimulated ATPase activity of purified HSP70-1A. (ii) A GST fusion protein comprising the C terminus of SERT pulled down HSP70-1A. The interaction between HSP70-1A and SERT was visualized in live cells by Förster resonance energy transfer: it was restricted to endoplasmic reticulum-resident transporters and enhanced by an inhibitor that traps HSP70-1A in its closed state. (iv) Co-immunoprecipitation confirmed complex formation of SERT with HSP70-1A and HSP90β. Consistent with an HSP relay, co-chaperones (e.g. HSC70-HSP90-organizing protein) were co-immunoprecipitated with the stalled mutants SERT-R607A/I608A and SERT-P601A/G602A. (v) Depletion of HSP90β by siRNA or its inhibition increased the cell surface expression of wild type SERT and SERT-F604Q. In contrast, SERT-R607A/I608A and SERT-P601A/G602A were only rendered susceptible to inhibition of HSP70 and HSP90 by concomitant pharmacochaperoning with noribogaine. (vi) In JAR cells, inhibition of HSP90 also increased the levels of SERT, indicating that endogenously expressed transporter was also susceptible to control by HSP90β. These findings support the concept that the folding trajectory of SERT is sampled by a cytoplasmic chaperone relay.

  3. Inhibition of Hsp70 by Methylene Blue Affects Signaling Protein Function and Ubiquitination and Modulates Polyglutamine Protein Degradation*


    Wang, Adrienne M; Morishima, Yoshihiro; Clapp, Kelly M.; Peng, Hwei-Ming; Pratt, William B.; Gestwicki, Jason E.; Osawa, Yoichi; Lieberman, Andrew P.


    The Hsp90/Hsp70-based chaperone machinery regulates the activity and degradation of many signaling proteins. Cycling with Hsp90 stabilizes client proteins, whereas Hsp70 interacts with chaperone-dependent E3 ubiquitin ligases to promote protein degradation. To probe these actions, small molecule inhibitors of Hsp70 would be extremely useful; however, few have been identified. Here we test the effects of methylene blue, a recently described inhibitor of Hsp70 ATPase activity, in three well est...

  4. CancerHSP: anticancer herbs database of systems pharmacology (United States)

    Tao, Weiyang; Li, Bohui; Gao, Shuo; Bai, Yaofei; Shar, Piar Ali; Zhang, Wenjuan; Guo, Zihu; Sun, Ke; Fu, Yingxue; Huang, Chao; Zheng, Chunli; Mu, Jiexin; Pei, Tianli; Wang, Yuan; Li, Yan; Wang, Yonghua


    The numerous natural products and their bioactivity potentially afford an extraordinary resource for new drug discovery and have been employed in cancer treatment. However, the underlying pharmacological mechanisms of most natural anticancer compounds remain elusive, which has become one of the major obstacles in developing novel effective anticancer agents. Here, to address these unmet needs, we developed an anticancer herbs database of systems pharmacology (CancerHSP), which records anticancer herbs related information through manual curation. Currently, CancerHSP contains 2439 anticancer herbal medicines with 3575 anticancer ingredients. For each ingredient, the molecular structure and nine key ADME parameters are provided. Moreover, we also provide the anticancer activities of these compounds based on 492 different cancer cell lines. Further, the protein targets of the compounds are predicted by state-of-art methods or collected from literatures. CancerHSP will help reveal the molecular mechanisms of natural anticancer products and accelerate anticancer drug development, especially facilitate future investigations on drug repositioning and drug discovery. CancerHSP is freely available on the web at PMID:26074488

  5. In silico identification of carboxylate clamp type tetratricopeptide repeat proteins in Arabidopsis and rice as putative co-chaperones of Hsp90/Hsp70.

    Directory of Open Access Journals (Sweden)

    Bishun D Prasad

    Full Text Available The essential eukaryotic molecular chaperone Hsp90 operates with the help of different co-chaperones, which regulate its ATPase activity and serve as adaptors to recruit client proteins and other molecular chaperones, such as Hsp70, to the Hsp90 complex. Several Hsp90 and Hsp70 co-chaperones contain the tetratricopeptide repeat (TPR domain, which interacts with the highly conserved EEVD motif at the C-terminal ends of Hsp90 and Hsp70. The acidic side chains in EEVD interact with a subset of basic residues in the TPR binding pocket called a 'carboxylate clamp'. Since the carboxylate clamp residues are conserved in the TPR domains of known Hsp90/Hsp70 co-chaperones, we carried out an in silico search for TPR proteins in Arabidopsis and rice comprising of at least one three-motif TPR domain with conserved amino acid residues required for Hsp90/Hsp70 binding. This approach identified in Arabidopsis a total of 36 carboxylate clamp (CC-TPR proteins, including 24 novel proteins, with potential to interact with Hsp90/Hsp70. The newly identified CC-TPR proteins in Arabidopsis and rice contain additional protein domains such as ankyrin, SET, octicosapeptide/Phox/Bem1p (Phox/PB1, DnaJ-like, thioredoxin, FBD and F-box, and protein kinase and U-box, indicating varied functions for these proteins. To provide proof-of-concept of the newly identified CC-TPR proteins for interaction with Hsp90, we demonstrated interaction of AtTPR1 and AtTPR2 with AtHsp90 in yeast two-hybrid and in vitro pull down assays. These findings indicate that the in silico approach used here successfully identified in a genome-wide context CC-TPR proteins with potential to interact with Hsp90/Hsp70, and further suggest that the Hsp90/Hsp70 system relies on TPR co-chaperones more than it was previously realized.

  6. Determination of the dose rapidity of a {sup 90} Sr beta radiation source using thermoluminescent dosemeters; Determinacion de la rapidez de dosis de una fuente de radiacion beta de {sup 90}Sr usando dosimetros termoluminiscentes

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez M, P.R.; Azorin N, J.; Rivera M, T. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)


    The thermoluminescent dosemeters developed in Mexico, have been used efficiently in environmental and personal dosimetry. When the dose rate of some source is not known can be estimated with the use of thermoluminescent dosemeters taking in account the geometrical array used in the irradiations for reproducibility of the results in posterior irradiations. In this work it was estimated the dose rate of a {sup 90} Sr-{sup 90} Y beta radiation source which is property of the Nuclear Sciences Institute, UNAM, therefore it was l ended to the Metropolitan Autonomous University- Iztapalapa Unit for the characterization of new Tl materials, taking account of the institutional collaboration agreements. (Author)

  7. 70 kD stress protein (Hsp70) analysis in living shallow-water benthic foraminifera

    Digital Repository Service at National Institute of Oceanography (India)

    Heinz, P.; Marten, R.A.; Linshy, V.N.; Haap, T.; Geslin, E.; Kohler, H-R.

    Hsp70 is a phylogenetically highly conserved protein family present in all eukaryotic organisms tested so far. Its synthesis is induced by proteotoxic stress. The detection of Hsp70 in foraminifera is presented here. We introduce a standard...

  8. RNAi knockdown of Hop (Hsp70/Hsp90 organising protein) decreases invasion via MMP-2 down regulation.

    LENUS (Irish Health Repository)

    Walsh, Naomi


    We previously identified Hop as over expressed in invasive pancreatic cancer cell lines and malignant tissues of pancreatic cancer patients, suggesting an important role for Hop in the biology of invasive pancreatic cancer. Hop is a co-chaperone protein that binds to both Hsp70\\/Hsp90. We hypothesised that by targeting Hop, signalling pathways modulating invasion and client protein stabilisation involving Hsp90-dependent complexes may be altered. In this study, we show that Hop knockdown by small interfering (si)RNA reduces the invasion of pancreatic cancer cells, resulting in decreased expression of the downstream target gene, matrix metalloproteinases-2 (MMP-2). Hop in conditioned media co-immunoprecipitates with MMP-2, implicating a possible extracellular function for Hop. Knockdown of Hop expression also reduced expression levels of Hsp90 client proteins, HER2, Bcr-Abl, c-MET and v-Src. Furthermore, Hop is strongly expressed in high grade PanINs compared to lower PanIN grades, displaying differential localisation in invasive ductal pancreatic cancer, indicating that the localisation of Hop is an important factor in pancreatic tumours. Our data suggests that the attenuation of Hop expression inactivates key signal transduction proteins which may decrease the invasiveness of pancreatic cancer cells possibly through the modulation of Hsp90 activity. Therefore, targeting Hop in pancreatic cancer may constitute a viable strategy for targeted cancer therapy.

  9. RNAi knockdown of Hop (Hsp70/Hsp90 organising protein) decreases invasion via MMP-2 down regulation. (United States)

    Walsh, Naomi; Larkin, AnneMarie; Swan, Niall; Conlon, Kevin; Dowling, Paul; McDermott, Ray; Clynes, Martin


    We previously identified Hop as over expressed in invasive pancreatic cancer cell lines and malignant tissues of pancreatic cancer patients, suggesting an important role for Hop in the biology of invasive pancreatic cancer. Hop is a co-chaperone protein that binds to both Hsp70/Hsp90. We hypothesised that by targeting Hop, signalling pathways modulating invasion and client protein stabilisation involving Hsp90-dependent complexes may be altered. In this study, we show that Hop knockdown by small interfering (si)RNA reduces the invasion of pancreatic cancer cells, resulting in decreased expression of the downstream target gene, matrix metalloproteinases-2 (MMP-2). Hop in conditioned media co-immunoprecipitates with MMP-2, implicating a possible extracellular function for Hop. Knockdown of Hop expression also reduced expression levels of Hsp90 client proteins, HER2, Bcr-Abl, c-MET and v-Src. Furthermore, Hop is strongly expressed in high grade PanINs compared to lower PanIN grades, displaying differential localisation in invasive ductal pancreatic cancer, indicating that the localisation of Hop is an important factor in pancreatic tumours. Our data suggests that the attenuation of Hop expression inactivates key signal transduction proteins which may decrease the invasiveness of pancreatic cancer cells possibly through the modulation of Hsp90 activity. Therefore, targeting Hop in pancreatic cancer may constitute a viable strategy for targeted cancer therapy.

  10. Sequence and domain conservation of the coelacanth Hsp40 and Hsp90 chaperones suggests conservation of function. (United States)

    Bishop, Özlem Tastan; Edkins, Adrienne Lesley; Blatch, Gregory Lloyd


    Molecular chaperones and their associated co-chaperones play an important role in preserving and regulating the active conformational state of cellular proteins. The chaperone complement of the Indonesian Coelacanth, Latimeria menadoensis, was elucidated using transcriptomic sequences. Heat shock protein 90 (Hsp90) and heat shock protein 40 (Hsp40) chaperones, and associated co-chaperones were focused on, and homologous human sequences were used to search the sequence databases. Coelacanth homologs of the cytosolic, mitochondrial and endoplasmic reticulum (ER) homologs of human Hsp90 were identified, as well as all of the major co-chaperones of the cytosolic isoform. Most of the human Hsp40s were found to have coelacanth homologs, and the data suggested that all of the chaperone machinery for protein folding at the ribosome, protein translocation to cellular compartments such as the ER and protein degradation were conserved. Some interesting similarities and differences were identified when interrogating human, mouse, and zebrafish homologs. For example, DnaJB13 is predicted to be a non-functional Hsp40 in humans, mouse, and zebrafish due to a corrupted histidine-proline-aspartic acid (HPD) motif, while the coelacanth homolog has an intact HPD. These and other comparisons enabled important functional and evolutionary questions to be posed for future experimental studies.

  11. Following the Expression of HSP 70 Gene by Semiquantification

    Directory of Open Access Journals (Sweden)

    Jana Tkáčová


    Full Text Available We conducted feeding experiment with broiler chickens Coob 500. We used lucerne meal in the feeding mixtures. It contains carotenoids including lycopen. Lycopene may have important antioxidant implications. Lycopene levels in serum or tissues do not correlate well with the overall intake. It is a potent antioxidant in vitro and in human studies. We followed the effect of lucerne meal on expression of protein HSP 70 in our experiment. HSP 70 belongs to large group of high shock proteins which mediate organism´s response not only to temperature rise, but also to other unfavorable factors, e.g. exposure to heavy metals, hormones, oxidants, etc. We isolated mRNA from heart, spleen, liver, abdominal fat and we transferred transcription and PCR. Expression of HSP 70 was visualized on agarose gel. We detected that the lowest expression was in the heart, the greatest expression was detected in the liver.

  12. HSP25 Protects Radiation-Induced Salivary Gland Damage

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hae June; Lee, Yoon Jin; Kwon, Hee Choong; Lee, Su Jae; Bae, Sang Woo; Lee, Yun Sil [Korea Institute of Radiological Medical Sciences, Seoul (Korea, Republic of); Kim, Sung Ho [Chonnam National Univ., Gwangju (Korea, Republic of)


    Irradiation (IR) is a central treatment modality administered for head and neck malignancies. A significant consequence of this IR treatment is irreversible damage to salivary gland in the IR field. While the exact mechanism of salivary gland damage remains enigmatic, fluid secreting acinar cells are lost, and saliva output is dramatically reduced. Previously we have reported that heat shock protein 25 (HSP25) induced radioresistance in vitro. HSP25 interferes negatively with apoptosis through several pathways which involve its direct interaction with cytochrome c, protein kinase c delta or Akt. And localized gene transfer to salivary glands has great potential for the treatment of salivary gland. Herein, we investigated whether HSP25 can use as radio protective molecules for radiation-induced salivary gland damage in vivo.

  13. The conformational dynamics of the mitochondrial Hsp70 chaperone. (United States)

    Mapa, Koyeli; Sikor, Martin; Kudryavtsev, Volodymyr; Waegemann, Karin; Kalinin, Stanislav; Seidel, Claus A M; Neupert, Walter; Lamb, Don C; Mokranjac, Dejana


    Heat shock proteins 70 (Hsp70) represent a ubiquitous and conserved family of molecular chaperones involved in a plethora of cellular processes. The dynamics of their ATP hydrolysis-driven and cochaperone-regulated conformational cycle are poorly understood. We used fluorescence spectroscopy to analyze, in real time and at single-molecule resolution, the effects of nucleotides and cochaperones on the conformation of Ssc1, a mitochondrial member of the family. We report that the conformation of its ADP state is unexpectedly heterogeneous, in contrast to a uniform ATP state. Substrates are actively involved in determining the conformation of Ssc1. The J protein Mdj1 does not interact transiently with the chaperone, as generally believed, but rather is released slowly upon ATP hydrolysis. Analysis of the major bacterial Hsp70 revealed important differences between highly homologous members of the family, possibly explaining tuning of Hsp70 chaperones to meet specific functions in different organisms and cellular compartments.

  14. Hsp90: A New Player in DNA Repair?

    Directory of Open Access Journals (Sweden)

    Rosa Pennisi


    Full Text Available Heat shock protein 90 (Hsp90 is an evolutionary conserved molecular chaperone that, together with Hsp70 and co-chaperones makes up the Hsp90 chaperone machinery, stabilizing and activating more than 200 proteins, involved in protein homeostasis (i.e., proteostasis, transcriptional regulation, chromatin remodeling, and DNA repair. Cells respond to DNA damage by activating complex DNA damage response (DDR pathways that include: (i cell cycle arrest; (ii transcriptional and post-translational activation of a subset of genes, including those associated with DNA repair; and (iii triggering of programmed cell death. The efficacy of the DDR pathways is influenced by the nuclear levels of DNA repair proteins, which are regulated by balancing between protein synthesis and degradation as well as by nuclear import and export. The inability to respond properly to either DNA damage or to DNA repair leads to genetic instability, which in turn may enhance the rate of cancer development. Multiple components of the DNA double strand breaks repair machinery, including BRCA1, BRCA2, CHK1, DNA-PKcs, FANCA, and the MRE11/RAD50/NBN complex, have been described to be client proteins of Hsp90, which acts as a regulator of the diverse DDR pathways. Inhibition of Hsp90 actions leads to the altered localization and stabilization of DDR proteins after DNA damage and may represent a cell-specific and tumor-selective radiosensibilizer. Here, the role of Hsp90-dependent molecular mechanisms involved in cancer onset and in the maintenance of the genome integrity is discussed and highlighted.

  15. Inhibition of Hsp27 Radiosensitizes Head-and-Neck Cancer by Modulating Deoxyribonucleic Acid Repair

    Energy Technology Data Exchange (ETDEWEB)

    Guttmann, David M.; Hart, Lori [Department of Radiation Oncology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania (United States); Du, Kevin [Department of Radiation Oncology, New York University School of Medicine, New York, New York (United States); Seletsky, Andrew [Department of Biology, Drexel University, Philadelphia, Pennsylvania (United States); Koumenis, Constantinos, E-mail: [Department of Radiation Oncology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania (United States)


    Purpose: To present a novel method of tumor radiosensitization through Hsp27 knockdown using locked nucleic acid (LNA) and to investigate the role of Hsp27 in DNA double strand break (DSB) repair. Methods and Materials: Clonogenic survival assays, immunoblotting, the proximity ligation assay, and γH2AX foci analysis were conducted in SQ20B and FaDu human head-and-neck cancer cell lines treated with Hsp27 LNA and Hsp27 short hairpin RNA (shRNA). Additionally, nude mice with FaDu flank tumors were treated with fractionated radiation therapy after pretreatment with Hsp27 LNA and monitored for tumor growth. Results: Hsp27 LNA and Hsp27 shRNA radiosensitized head-and-neck cancer cell lines in an Hsp27-dependent manner. Ataxia-Telangectasia Mutated-mediated DNA repair signaling was impaired in irradiated cells with Hsp27 knockdown. ATM kinase inhibition abrogated the radiosensitizing effect of Hsp27. Furthermore, Hsp27 LNA and shRNA both attenuated DNA repair kinetics after radiation, and Hsp27 was found to colocalize with ATM in both untreated and irradiated cells. Last, combined radiation and Hsp27 LNA treatment in tumor xenografts in nude mice suppressed tumor growth compared with either treatment alone. Conclusions: These results support a radiosensitizing property of Hsp27 LNA in vitro and in vivo, implicate Hsp27 in double strand break repair, and suggest that Hsp27 LNA might eventually serve as an effective clinical agent in the radiotherapy of head-and-neck cancer.

  16. Hsp20 functions as a novel cardiokine in promoting angiogenesis via activation of VEGFR2.

    Directory of Open Access Journals (Sweden)

    Xiaowei Zhang

    Full Text Available Heat shock proteins (Hsps are well appreciated as intrinsic protectors of cardiomyocytes against numerous stresses. Recent studies have indicated that Hsp20 (HspB6, a small heat shock protein, was increased in blood from cardiomyopathic hamsters. However, the exact source of the increased circulating Hsp20 and its potential role remain obscure. In this study, we observed that the circulating Hsp20 was increased in a transgenic mouse model with cardiac-specific overexpression of Hsp20, compared with wild-type mice, suggesting its origin from cardiomyocytes. Consistently, culture media harvested from Hsp20-overexpressing cardiomyocytes by Ad.Hsp20 infection contained an increased amount of Hsp20, compared to control media. Furthermore, we identified that Hsp20 was secreted through exosomes, independent of the endoplasmic reticulum-Golgi pathway. To investigate whether extracellular Hsp20 promotes angiogenesis, we treated human umbilical vein endothelial cells (HUVECs with recombinant human Hsp20 protein, and observed that Hsp20 dose-dependently promoted HUVEC proliferation, migration and tube formation. Moreover, a protein binding assay and immunostaining revealed an interaction between Hsp20 and VEGFR2. Accordingly, stimulatory effects of Hsp20 on HUVECs were blocked by a VEGFR2 neutralizing antibody and CBO-P11 (a VEGFR inhibitor. These in vitro data are consistent with the in vivo findings that capillary density was significantly enhanced in Hsp20-overexpressing hearts, compared to non-transgenic hearts. Collectively, our findings demonstrate that Hsp20 serves as a novel cardiokine in regulating myocardial angiogenesis through activation of the VEGFR signaling cascade.

  17. Epigallocatechin-3-gallate suppresses the expression of HSP70 and HSP90 and exhibits anti-tumor activity in vitro and in vivo

    Directory of Open Access Journals (Sweden)

    Yoon Jung-Hoon


    Full Text Available Abstract Background Epigallocatechin-3-gallate (EGCG, one of the major catechins in green tea, is a potential chemopreventive agent for various cancers. The aim of this study was to examine the effect of EGCG on the expression of heat shock proteins (HSPs and tumor suppression. Methods Cell colony formation was evaluated by a soft agar assay. Transcriptional activity of HSP70 and HSP90 was determined by luciferase reporter assay. An EGCG-HSPs complex was prepared using EGCG attached to the cyanogen bromide (CNBr-activated Sepharose 4B. In vivo effect of EGCG on tumor growth was examined in a xenograft model. Results Treatment with EGCG decreased cell proliferation and colony formation of MCF-7 human breast cancer cells. EGCG specifically inhibited the expression of HSP70 and HSP90 by inhibiting the promoter activity of HSP70 and HSP90. Pretreatment with EGCG increased the stress sensitivity of MCF-7 cells upon heat shock (44°C for 1 h or oxidative stress (H2O2, 500 μM for 24 h. Moreover, treatment with EGCG (10 mg/kg in a xenograft model resulted in delayed tumor incidence and reduced tumor size, as well as the inhibition of HSP70 and HSP90 expression. Conclusions Overall, these findings demonstrate that HSP70 and HSP90 are potent molecular targets of EGCG and suggest EGCG as a drug candidate for the treatment of human cancer.

  18. Ratio of phosphorylated HSP27 to nonphosphorylated HSP27 biphasically acts as a determinant of cellular fate in gemcitabine-resistant pancreatic cancer cells. (United States)

    Kang, Dongxu; Choi, Hye Jin; Kang, Sujin; Kim, So Young; Hwang, Yong-Sic; Je, Suyeon; Han, Zhezhu; Kim, Joo-Hang; Song, Jae J


    Gemcitabine has been used most commonly as an anticancer drug to treat advanced pancreatic cancer patients. However, intrinsic or acquired resistance of pancreatic cancer to gemcitabine was also developed, which leads to very low five-year survival rates. Here, we investigated whether cellular levels of HSP27 phosphorylation act as a determinant of cellular fate with gemcitabine. In addition we have demonstrated whether HSP27 downregulation effectively could overcome the acquisition of gemcitabine resistance by using transcriptomic analysis. We observed that gemcitabine induced p38/HSP27 phosphorylation and caused acquired resistance. After acquisition of gemcitabine resistance, cancer cells showed higher activity of NF-κB. NF-κB activity, as well as colony formation in gemcitabine-resistant pancreatic cancer cells, was significantly decreased by HSP27 downregulation and subsequent TRAIL treatment, showing that HSP27 was a common network mediator of gemcitabine/TRAIL-induced cell death. After transcriptomic analysis, gene fluctuation after HSP27 downregulation was very similar to that of pancreatic cancer cells susceptible to gemcitabine, and then in opposite position to that of acquired gemcitabine resistance, which makes it possible to downregulate HSP27 to overcome the acquired gemcitabine resistance to function as an overall survival network inhibitor. Most importantly, we demonstrated that the ratio of phosphorylated HSP27 to nonphosphorylated HSP27 rather than the cellular level of HSP27 itself acts biphasically as a determinant of cellular fate in gemcitabine-resistant pancreatic cancer cells.

  19. Effect of ATP on the release of hsp 70 and hsp 40 from the nucleus in heat-shocked HeLa cells. (United States)

    Ohtsuka, K; Utsumi, K R; Kaneda, T; Hattori, H


    We have recently found a novel 40-kDa heat-shock protein (hsp 40) in mammalian and avian cells and reported that the N-terminal amino acid sequence of mammalian hsp 40 has homology with the bacterial DnaJ heat-shock protein. Also, hsp 40 has been shown to be translocated from the cytoplasm into the nuclei/nucleoli by heat shock and colocalized with hsc 70 (p73) in the nucleoli of exactly the same cells. We here investigated the effect of ATP on the release of hsp 70 (both constitutive p73 and inducible p72) and hsp 40 from the nuclei/nucleoli of heat-shocked HeLa cells which were permeabilized with Nonidet-P40 using immunofluorescence and immunoblotting. Hsp 70 in the nucleoli was released by the addition of ATP but not by ADP, GTP, nonhydrolyzable ATP, nor high salt buffer. In contrast, hsp 40 was not released from the nucleoli with any of these treatments or any combination of these treatments. Thus, hsp 40 might dissociate spontaneously from the nucleoli after hsp 70 has been released in an ATP-dependent manner. Using cell fractionation methods, we showed that while the majority of hsp 40 is localized in the cytoplasm, a small portion of it is located in the microsome fraction in non-heat-shocked control cells and in cells which recovered from heat shock.

  20. Stability of the Human Hsp90-p50Cdc37 Chaperone Complex against Nucleotides and Hsp90 Inhibitors, and the Influence of Phosphorylation by Casein Kinase 2

    Directory of Open Access Journals (Sweden)

    Sanne H. Olesen


    Full Text Available The molecular chaperone Hsp90 is regulated by co-chaperones such as p50Cdc37, which recruits a wide selection of client protein kinases. Targeted disruption of the Hsp90-p50Cdc37 complex by protein–protein interaction (PPI inhibitors has emerged as an alternative strategy to treat diseases characterized by aberrant Hsp90 activity. Using isothermal microcalorimetry, ELISA and GST-pull down assays we evaluated reported Hsp90 inhibitors and nucleotides for their ability to inhibit formation of the human Hsp90β-p50Cdc37 complex, reconstituted in vitro from full-length proteins. Hsp90 inhibitors, including the proposed PPI inhibitors gedunin and H2-gamendazole, did not affect the interaction of Hsp90 with p50Cdc37 in vitro. Phosphorylation of Hsp90 and p50Cdc37 by casein kinase 2 (CK2 did not alter the thermodynamic signature of complex formation. However, the phosphorylated complex was vulnerable to disruption by ADP (IC50 = 32 µM, while ATP, AMPPNP and Hsp90 inhibitors remained largely ineffective. The differential inhibitory activity of ADP suggests that phosphorylation by CK2 primes the complex for dissociation in response to a drop in ATP/ADP levels. The approach applied herein provides robust assays for a comprehensive biochemical evaluation of potential effectors of the Hsp90-p50Cdc37 complex, such as phosphorylation by a kinase or the interaction with small molecule ligands.

  1. Stability of the human Hsp90-p50Cdc37 chaperone complex against nucleotides and Hsp90 inhibitors, and the influence of phosphorylation by casein kinase 2. (United States)

    Olesen, Sanne H; Ingles, Donna J; Zhu, Jin-Yi; Martin, Mathew P; Betzi, Stephane; Georg, Gunda I; Tash, Joseph S; Schönbrunn, Ernst


    The molecular chaperone Hsp90 is regulated by co-chaperones such as p50Cdc37, which recruits a wide selection of client protein kinases. Targeted disruption of the Hsp90-p50Cdc37 complex by protein-protein interaction (PPI) inhibitors has emerged as an alternative strategy to treat diseases characterized by aberrant Hsp90 activity. Using isothermal microcalorimetry, ELISA and GST-pull down assays we evaluated reported Hsp90 inhibitors and nucleotides for their ability to inhibit formation of the human Hsp90β-p50Cdc37 complex, reconstituted in vitro from full-length proteins. Hsp90 inhibitors, including the proposed PPI inhibitors gedunin and H2-gamendazole, did not affect the interaction of Hsp90 with p50Cdc37 in vitro. Phosphorylation of Hsp90 and p50Cdc37 by casein kinase 2 (CK2) did not alter the thermodynamic signature of complex formation. However, the phosphorylated complex was vulnerable to disruption by ADP (IC50 = 32 µM), while ATP, AMPPNP and Hsp90 inhibitors remained largely ineffective. The differential inhibitory activity of ADP suggests that phosphorylation by CK2 primes the complex for dissociation in response to a drop in ATP/ADP levels. The approach applied herein provides robust assays for a comprehensive biochemical evaluation of potential effectors of the Hsp90-p50Cdc37 complex, such as phosphorylation by a kinase or the interaction with small molecule ligands.

  2. Stability of the Human Hsp90-p50Cdc37 Chaperone Complex against Nucleotides and Hsp90 Inhibitors, and the Influence of Phosphorylation by Casein Kinase 2 (United States)

    Olesen, Sanne H.; Ingles, Donna J.; Zhu, Jin-Yi; Martin, Mathew P.; Betzi, Stephane; Georg, Gunda I.; Tash, Joseph S.; Schönbrunn, Ernst


    The molecular chaperone Hsp90 is regulated by co-chaperones such as p50Cdc37, which recruits a wide selection of client protein kinases. Targeted disruption of the Hsp90-p50Cdc37 complex by protein-protein interaction (PPI) inhibitors has emerged as an alternative strategy to treat diseases characterized by aberrant Hsp90 activity. Using isothermal microcalorimetry, ELISA and GST-pull down assays we evaluated reported Hsp90 inhibitors and nucleotides for their ability to inhibit formation of the human Hsp90β-p50Cdc37 complex, reconstituted in-vitro from full-length proteins. Hsp90 inhibitors, including the proposed PPI inhibitors gedunin and H2-gamendazole, did not affect the interaction of Hsp90 with p50Cdc37 in vitro. Phosphorylation of Hsp90 and p50Cdc37 by casein kinase 2 (CK2) did not alter the thermodynamic signature of complex formation. However, the phosphorylated complex was vulnerable to disruption by ADP (IC50 = 32 µM), while ATP, AMPPNP and Hsp90 inhibitors remained largely ineffective. The differential inhibitory activity of ADP suggests that phosphorylation by CK2 primes the complex for dissociation in response to a drop in ATP/ADP levels. The approach applied herein provides robust assays for a comprehensive biochemical evaluation of potential effectors of the Hsp90-p50Cdc37 complex, such as phosphorylation by a kinase or the interaction with small molecule ligands. PMID:25608045

  3. Constitutively-active androgen receptor variants function independently of the HSP90 chaperone but do not confer resistance to HSP90 inhibitors. (United States)

    Gillis, Joanna L; Selth, Luke A; Centenera, Margaret M; Townley, Scott L; Sun, Shihua; Plymate, Stephen R; Tilley, Wayne D; Butler, Lisa M


    The development of lethal, castration resistant prostate cancer is associated with adaptive changes to the androgen receptor (AR), including the emergence of mutant receptors and truncated, constitutively active AR variants. AR relies on the molecular chaperone HSP90 for its function in both normal and malignant prostate cells, but the requirement for HSP90 in environments with aberrant AR expression is largely unknown. Here, we investigated the efficacy of three HSP90 inhibitors, 17-AAG, HSP990 and AUY922, against clinically-relevant AR missense mutants and truncated variants. HSP90 inhibition effectively suppressed the signaling of wild-type AR and all AR missense mutants tested. By contrast, two truncated AR variants, AR-V7 and ARv567es, exhibited marked resistance to HSP90 inhibitors. Supporting this observation, nuclear localization of the truncated AR variants was not affected by HSP90 inhibition and AR variant:HSP90 complexes could not be detected in prostate cancer cells. Interestingly, HSP90 inhibition resulted in accumulation of AR-V7 and ARv567es in both cell lines and human tumor explants. Despite the apparent independence of AR variants from HSP90 and their treatment-associated induction, the growth of cell lines with endogenous or enforced expression of AR-V7 or ARv567es remained highly sensitive to AUY922. This study demonstrates that functional AR variant signaling does not confer resistance to HSP90 inhibition, yields insight into the interaction between AR and HSP90 and provides further impetus for the clinical application of HSP90 inhibitors in advanced prostate cancer.

  4. FLZ, a novel HSP27 and HSP70 inducer, protects SH-SY5Y cells from apoptosis caused by MPP(+). (United States)

    Kong, Xiang-Chen; Zhang, Dan; Qian, Cheng; Liu, Geng-Tao; Bao, Xiu-Qi


    Heat shock proteins (HSPs) play an essential role in various neurodegenerative diseases. Manipulation of upregulation of HSPs in cells has been demonstrated to provide a therapeutic strategy to counteract the misfolding and aggregation of proteins that resulted in neurodegenerative disease. Our previous studies have shown that FLZ, a synthetic novel derivative of squamosamide from a Chinese herb, had potent neuroprotective effect against several experimental Parkinson's disease (PD) models. However, the mechanism of its neuroprotective effect is still not clarified. The present study demonstrated that FLZ induced HSP27 and HSP70 proteins and mRNA expression in a time- and dose-dependent manner in SH-SY5Y cells. Further studies showed that FLZ treatment stimulated the activation of heat shock factor 1 (HSF1) and its regulatory kinase Akt. Inactivation of Akt pathway by the PI3K inhibitor LY294002 blocked the expression of HSP27 and HSP70 induced by FLZ. Moreover, the inducing effects of FLZ on HSP27, HSP70, and HSF1 were all blocked by quercetin, an inhibitor of HSP biosynthesis. The cytoprotective effect of HSP27/HSP70 induced by FLZ against MPP(+) was assessed in SH-SY5Y cells. The pretreatment of FLZ significantly induced the accumulations of HSP27/HSP70 and suppressed the apoptosis caused by MPP(+) in SH-SY5Y cells. However, the protective effects of FLZ against MPP(+) were significantly blocked by quercetin, which indicated that the cytoprotective action of FLZ against MPP(+)-induced apoptosis is at least partially mediated by its induction of HSP27/HSP70. These results provide new evidence for elucidating the mechanism of the neuroprotective effect of FLZ against PD.

  5. Neonatal Death and Heart Failure in Mouse with Transgenic HSP60 Expression

    Directory of Open Access Journals (Sweden)

    Tsung-Hsien Chen


    Full Text Available Mitochondrial heat shock proteins, such as HSP60, are chaperones responsible for the folding, transport, and quality control of mitochondrial matrix proteins and are essential for maintaining life. Both prosurvival and proapoptotic roles have been proposed for HSP60, and HSP60 is reportedly involved in the initiation of autoimmune, metabolic, and cardiovascular diseases. The role of HSP60 in pathogenesis of these diseases remains unclear, partly because of the lack of mouse models expressing HSP60. In this study we generated HSP60 conditional transgenic mice suitable for investigating in vivo outcomes by expressing HSP60 at the targeted organ in disease models. Ubiquitous HSP60 induction in the embryonic stage caused neonatal death in mice at postnatal day 1. A high incidence of atrial septal defects was observed in HSP60-expressing mice, with increased apoptosis and myocyte degeneration that possibly contributed to massive hemorrhage and sponge-like cardiac muscles. Our results showed that neonatal heart failure through HSP60 induction likely involves developmental defects and excessive apoptosis. The conditional HSP60 mouse model is useful for studying crucial biological questions concerning HSP60.

  6. Investigating the Chaperone Properties of a Novel Heat Shock Protein, Hsp70.c, from Trypanosoma brucei

    Directory of Open Access Journals (Sweden)

    Adélle Burger


    Full Text Available The neglected tropical disease, African Trypanosomiasis, is fatal and has a crippling impact on economic development. Heat shock protein 70 (Hsp70 is an important molecular chaperone that is expressed in response to stress and Hsp40 acts as its co-chaperone. These proteins play a wide range of roles in the cell and they are required to assist the parasite as it moves from a cold blooded insect vector to a warm blooded mammalian host. A novel cytosolic Hsp70, from Trypanosoma brucei, TbHsp70.c, contains an acidic substrate binding domain and lacks the C-terminal EEVD motif. The ability of a cytosolic Hsp40 from Trypanosoma brucei J protein 2, Tbj2, to function as a co-chaperone of TbHsp70.c was investigated. The main objective was to functionally characterize TbHsp70.c to further expand our knowledge of parasite biology. TbHsp70.c and Tbj2 were heterologously expressed and purified and both proteins displayed the ability to suppress aggregation of thermolabile MDH and chemically denatured rhodanese. ATPase assays revealed a 2.8-fold stimulation of the ATPase activity of TbHsp70.c by Tbj2. TbHsp70.c and Tbj2 both demonstrated chaperone activity and Tbj2 functions as a co-chaperone of TbHsp70.c. In vivo heat stress experiments indicated upregulation of the expression levels of TbHsp70.c.

  7. BioFET-SIM

    DEFF Research Database (Denmark)

    Hediger, M. R.; Martinez, K. L.; Nygård, J.


    signals through its web-based interface The model also allows for predictions of the effects of changes in the experimental setup on the sensor signal. After an introduction to nanowire-based FET biosensors, this chapter reviews the theoretical basis of BioFET-SIM models describing both...... single and multiple charges on the analyte. Afterwards the usage of the interface and its relative command line version is briefly shown. Finally, possible applications of the BioFET-SIM model are presented. Among the possible uses of the interface, the effects on the predicted signal of pH, buffer ionic...... strength, analyte concentration, and analyte relative orientation on nanowire surface are illustrated. Wherever possible, a comparison to experimental data available in literature is given, displaying the potential of BioFETSIM for interpreting experimental results....

  8. Engineering BioBrick vectors from BioBrick parts


    Knight Thomas F; Endy Drew; Shetty Reshma P


    Abstract Background The underlying goal of synthetic biology is to make the process of engineering biological systems easier. Recent work has focused on defining and developing standard biological parts. The technical standard that has gained the most traction in the synthetic biology community is the BioBrick standard for physical composition of genetic parts. Parts that conform to the BioBrick assembly standard are BioBrick standard biological parts. To date, over 2,000 BioBrick parts have ...

  9. Expression of Hsp27 and Hsp70 in Lymphocytes and Plasma in Healthy Workers and Coal Miners with Lung Cancer

    Institute of Scientific and Technical Information of China (English)

    王海椒; 邢景才; 王峰; 韩文慧; 任侯卯; 邬堂春; 陈卫红


    In coal mines, main occupational hazard is coal-mine dust, which can cause health problem including coal workers' pneumoconiosis and lung cancer. Some heat shock proteins (Hsps) have been reported as an acute response to a wide variety of stressful stimuli. Whether Hsps protect against chronic environmental coal-mine dust over years is unknown. It is also interesting to know that whether the expression of Hsp27 and Hsp70 proteins as a marker for exposure is associated risk of lung cancer among coal miners. ...

  10. Energy and directional response for the Harshaw dosemeter holders 8814 and 8891, and its effect on the appropriate radiation qualities for absolute calibration. (United States)

    Børretzen, I; Wøhni, T


    The personal dosimetry laboratory at the Norwegian radiation protection authority utilises a two-element dosemeter card for measuring Hp(10) and Hp(0.07), in a Harshaw dosemeter holder type 8814. Energy and directional responses for photons and betas for this holder have been assessed, as well as for the new Harshaw holder type 8891. The energy response characteristics for the 12-1250 keV photon energy range, in terms of TL output per unit Hp(10) and Hp(0.07) have been evaluated. The maximum over-response to under-response ratio for the Hp(10) element was found to be 1.46 for the new type 8891 holder, as compared to 1.55 for the older type 8814. The new holder also displays a more favourable directional response for this element. For the Hp(0.07) element, no significant differences with regard to energy or directional responses were found. Selecting radiation energy for absolute calibration of the Hp(10) and Hp(0.07) elements are discussed.

  11. Energy and directional response for the Harshaw dosemeter holders 8814 and 8891, and its effect on the appropriate radiation qualities for absolute calibration

    Energy Technology Data Exchange (ETDEWEB)

    Borretzen, I.; Wohni, T


    The personal dosimetry laboratory at the Norwegian radiation protection authority utilises a two-element dosemeter card for measuring H{sub p}(10) and H{sub p}(0.07), in a Harshaw dosemeter holder type 8814. Energy and directional responses for photons and betas for this holder have been assessed, as well as for the new Harshaw holder type 8891. The energy response characteristics for the 12-1250 keV photon energy range, in terms of TL output per unit H{sub p}(10) and H{sub p}(0.07) have been evaluated. The maximum over-response to under-response ratio for the H{sub p}(10) element was found to be 1.46 for the new type 8891 holder, as compared to 1.55 for the older type 8814. The new holder also displays a more favourable directional response for this element. For the H{sub p}(0.07) element, no significant differences with regard to energy or directional responses were found. Selecting radiation energy for absolute calibration of the H{sub p}(10) and H{sub p}(0.07) elements are discussed. (author)

  12. Spatio-temporal regulation of Hsp90-ligand complex leads to immune activation.

    Directory of Open Access Journals (Sweden)

    Yasuaki eTamura


    Full Text Available Hsp90 is the most abundant cytosolic HSP and is known to act as a molecular chaperone. We found that an Hsp90-cancer antigen peptide complex was efficiently cross-presented by human monocyte-derived dendritic cells and induced peptide-specific cytotoxic T lymphocytes. Furthermore, we observed that the internalized Hsp90-peptide complex was strictly sorted to the Rab5+, EEA1+ static early endosome and the Hsp90-chaperoned peptide was processed and bound to MHC class I molecules through a endosome-recycling pathway. We also found that extracellular Hsp90 complexed with CpG-A or self-DNA stimulates production of a large amount of IFN-α from pDCs via static early endosome targeting. Thus, extracellular Hsp90 can target the antigen or nucleic acid to a static early endosome by spatio-temporal regulation. Moreover, we showed that Hsp90 associates with and delivers TLR7/9 from the ER to early endosomes for ligand recognition. Hsp90 inhibitor, geldanamycin derivative inhibited the Hsp90 association with TLR7/9, resulting in inhibition IFN-α production, leading to improvement of SLE symptoms. Interstingly, we observed that serum Hsp90 is clearly increased in patients with active SLE compared with that in patients with inactive disease. Serum Hsp90 detected in SLE patients binds to self-DNA and/or anti-DNA Ab, thus leading to stimulation of pDCs to produce IFN-α. Thus, Hsp90 plays a crucial role in the pathogenesis of SLE and that an Hsp90 inhibitor will therefore provide a new therapeutic approach to SLE and other nucleic acid-related autoimmune diseases. We will discuss how spatio-temporal regulation of Hsp90-ligand complexes within antigen-presenting cells affects the innate immunity and adaptive immunity.

  13. Specific Binding of Tetratricopeptide Repeat Proteins to Heat Shock Protein 70 (Hsp70) and Heat Shock Protein 90 (Hsp90) Is Regulated by Affinity and Phosphorylation. (United States)

    Assimon, Victoria A; Southworth, Daniel R; Gestwicki, Jason E


    Heat shock protein 70 (Hsp70) and heat shock protein 90 (Hsp90) require the help of tetratricopeptide repeat (TPR) domain-containing cochaperones for many of their functions. Each monomer of Hsp70 or Hsp90 can interact with only a single TPR cochaperone at a time, and each member of the TPR cochaperone family brings distinct functions to the complex. Thus, competition for TPR binding sites on Hsp70 and Hsp90 appears to shape chaperone activity. Recent structural and biophysical efforts have improved our understanding of chaperone-TPR contacts, focusing on the C-terminal EEVD motif that is present in both chaperones. To better understand these important protein-protein interactions on a wider scale, we measured the affinity of five TPR cochaperones, CHIP, Hop, DnaJC7, FKBP51, and FKBP52, for the C-termini of four members of the chaperone family, Hsc70, Hsp72, Hsp90α, and Hsp90β, in vitro. These studies identified some surprising selectivity among the chaperone-TPR pairs, including the selective binding of FKBP51/52 to Hsp90α/β. These results also revealed that other TPR cochaperones are only able to weakly discriminate between the chaperones or between their paralogs. We also explored whether mimicking phosphorylation of serine and threonine residues near the EEVD motif might impact affinity and found that pseudophosphorylation had selective effects on binding to CHIP but not other cochaperones. Together, these findings suggest that both intrinsic affinity and post-translational modifications tune the interactions between the Hsp70 and Hsp90 proteins and the TPR cochaperones.

  14. Comparative genomics and evolution of the HSP90 family of genes across all kingdoms of organisms

    Directory of Open Access Journals (Sweden)

    Zhong Daibin


    Full Text Available Abstract Background HSP90 proteins are essential molecular chaperones involved in signal transduction, cell cycle control, stress management, and folding, degradation, and transport of proteins. HSP90 proteins have been found in a variety of organisms suggesting that they are ancient and conserved. In this study we investigate the nuclear genomes of 32 species across all kingdoms of organisms, and all sequences available in GenBank, and address the diversity, evolution, gene structure, conservation and nomenclature of the HSP90 family of genes across all organisms. Results Twelve new genes and a new type HSP90C2 were identified. The chromosomal location, exon splicing, and prediction of whether they are functional copies were documented, as well as the amino acid length and molecular mass of their polypeptides. The conserved regions across all protein sequences, and signature sequences in each subfamily were determined, and a standardized nomenclature system for this gene family is presented. The proeukaryote HSP90 homologue, HTPG, exists in most Bacteria species but not in Archaea, and it evolved into three lineages (Groups A, B and C via two gene duplication events. None of the organellar-localized HSP90s were derived from endosymbionts of early eukaryotes. Mitochondrial TRAP and endoplasmic reticulum HSP90B separately originated from the ancestors of HTPG Group A in Firmicutes-like organisms very early in the formation of the eukaryotic cell. TRAP is monophyletic and present in all Animalia and some Protista species, while HSP90B is paraphyletic and present in all eukaryotes with the exception of some Fungi species, which appear to have lost it. Both HSP90C (chloroplast HSP90C1 and location-undetermined SP90C2 and cytosolic HSP90A are monophyletic, and originated from HSP90B by independent gene duplications. HSP90C exists only in Plantae, and was duplicated into HSP90C1 and HSP90C2 isoforms in higher plants. HSP90A occurs across all

  15. Effect of some parameters in the response of the Perspex 3042, Lot L amber dosemeters; Efectos de algunos parametros en la respuesta de los dosimetros ambar Perspex 3042, Lote L

    Energy Technology Data Exchange (ETDEWEB)

    Prieto M, E.F.; Barrera G, G. [Centro de Aplicaciones Tecnologicas y Desarrollo Nuclear (CEADEN). Calle 30, No. 502, esq. 5ta. Avenida, Playa, Ciudad de La Habana (Cuba)]. e-mail:


    The answer of the dosimetric systems is affected by several factors, for what should know as these factors they influence in each one of the different dosimetric systems and by this way to minimize its effect in the value of the absorbed dose and to obtain exact dose values. One of the dosimetric systems more used in the high dose dosimetry like routine dosemeter for the control of the irradiation process are the Perspex dosemeters, for their speed in the obtaining the information, their easy manipulation and the precision that they present. To this dosemeters group they belong the same as the Red and Clear the Amber, which are adequate for the measurement of the radiation dose in the range of high doses. The objective of the present work is to obtain the calibration curves of the dosemeters Amber Perspex 3042, Lot L under our work conditions, like they are the irradiation temperatures and of storage and to know the influence of the rate dose in the value of the absorbed dose for different measurement wavelengths, as well as, the relationship among the one post-irradiation time and the specific absorbance value induced in function of the absorbed dose. (Author)

  16. BioMEMS

    CERN Document Server

    Urban, Gerald A


    Explosive growth in the field of Microsystem Technology has introduced a variety of promising products in major disciplines from microelectronics to life sciences. 'Biomes' is a discipline which focuses on microsystems for living systems. This work presents the exciting field of bio-microsystems.

  17. Bio-Culturalism

    DEFF Research Database (Denmark)

    Grodal, Torben Kragh


    The article argues on the basis of analyses of successful films for children that not only cultural determinants but also innate determinats are important, and that film studies should combine cultural studies with cognitive theory, evolutionary theory and neuroscience, an approach that is called...... Bio-culturalism....

  18. Bio-technologies; Biotechnologies

    Energy Technology Data Exchange (ETDEWEB)

    Grawitz, X. [Systems Bio Industries, 92 - Boulogne Billancourt (France)


    This paper is a series of transparencies which describes the measures taken by Systems Bio-Industries company to adapt its central heating plants, turbines, engines and dryers to the new French 2910 by-law about thermal efficiency and environmental impact of heating plants. The project of development of a cogeneration system in the Angouleme site is briefly described. (J.S.)

  19. Bos indicus cattle possess greater basal concentrations of HSP27, alpha B-crystallin, and HSP70 in skeletal muscle in vivo compared with cattle. (United States)

    Mullins, C R; Zerby, H N; Fitzpatrick, L A; Parker, A J


    The objectives of the present study were to evaluate the basal concentrations of heat shock proteins (HSP) between and cattle and to determine if HSP basal concentrations change as an animal matures. A total of 40 cattle were used in a 2 × 2 factorial design to evaluate the effects of genotype and age (heifers and mature cows) on basal concentrations of heat shock protein 27 (HSP27), α B-crystallin (Cryab), and heat shock protein 70 (HSP70). Each experimental group of 10 animals was sampled on a separate day over a period of 4 wk during July 2014. A muscle sample was collected from the longissimus thoracis (LT) and concentrations of HSP were quantified using ELISA. There were no significant differences in HSP concentration for the interaction between age and genotype or for age alone. cattle had greater ( cattle. The results of this study show that basal in vivo HSP concentrations differ between and cattle. However, further studies are needed to investigate the relationship between HSP concentrations and meat tenderness with respect to genotypes to see if HSP concentrations account for at least some variability in tenderness differences.

  20. Study on routines and procedures of calibration of clinical dosemeter on gamma radiation beams at the IPEN-CNEN/SP, Brazil; Estudo das rotinas e procedimentos de calibracao de dosimetros clinicos em feixes de radiacao gama no IPEN-CNEN/SP

    Energy Technology Data Exchange (ETDEWEB)

    Damatto, Willian B.; Potiens, Maria da Penha A.; Santos, Gelson P.; Vivolo, Vitor, E-mail: wbdamatto@ipen.b, E-mail: mppalbu@ipen.b, E-mail: gpsantos@ipen.b, E-mail: vivolo@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)


    The calibration consists in the comparison of the equipment to be calibrated with other equipment already calibrated by another standard, which means the equipment that will accomplish shall be considered secondary or tertiary, depending on the calibration that will be accomplished. As the clinical dosemeters are used for quality control services in the hospitals, these dosemeters need a reliable measurement precision. Therefore, this work intends to demonstrate the importance of the clinical dosemeter and the calibration, requiring special care face to all clinical set

  1. The Double-Edged Sword: Conserved Functions of Extracellular Hsp90 in Wound Healing and Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Hance, Michael W.; Nolan, Krystal D.; Isaacs, Jennifer S., E-mail: [Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Hollings Cancer Center, Charleston, SC 29412 (United States)


    Heat shock proteins (Hsps) represent a diverse group of chaperones that play a vital role in the protection of cells against numerous environmental stresses. Although our understanding of chaperone biology has deepened over the last decade, the “atypical” extracellular functions of Hsps have remained somewhat enigmatic and comparatively understudied. The heat shock protein 90 (Hsp90) chaperone is a prototypic model for an Hsp family member exhibiting a duality of intracellular and extracellular functions. Intracellular Hsp90 is best known as a master regulator of protein folding. Cancers are particularly adept at exploiting this function of Hsp90, providing the impetus for the robust clinical development of small molecule Hsp90 inhibitors. However, in addition to its maintenance of protein homeostasis, Hsp90 has also been identified as an extracellular protein. Although early reports ascribed immunoregulatory functions to extracellular Hsp90 (eHsp90), recent studies have illuminated expanded functions for eHsp90 in wound healing and cancer. While the intended physiological role of eHsp90 remains enigmatic, its evolutionarily conserved functions in wound healing are easily co-opted during malignancy, a pathology sharing many properties of wounded tissue. This review will highlight the emerging functions of eHsp90 and shed light on its seemingly dichotomous roles as a benevolent facilitator of wound healing and as a sinister effector of tumor progression.

  2. Thermotolerance and HSP70 expression in the Mediterranean fruit fly Ceratitis capitata. (United States)

    Kalosaka, Katerina; Soumaka, Elisavet; Politis, Nikos; Mintzas, Anastassios C


    The relationship between Hsp70 expression and thermotolerance has been well documented in Drosophila melanogaster. However, there is limited information on this relationship in other insect species. In this report we describe the Hsp70-thermotolerance relationship in one of the major fruit fly pests, Ceratitis capitata (medfly). Hsp70 expression and thermotolerance were assayed at a range of temperatures in several stages of medfly development. The most thermotolerant stage was found to be the late larval stage (100% survival at 41 degrees C) followed by adult flies and late embryos (100% survival at 39 degrees C). These three stages showed a positive relationship between Hsp70 expression and thermotolerance. Mid-larval and mid-embryonic stages were found less thermotolerant and the Hsp70-thermotolerance relationship was not evident. Early embryos did not express Hsp70 at any temperature and exhibited the lowest thermotolerance. The relationship between Hsp70 and inducible thermotolerance was also studied in late larvae. A pretreatment at 37-39 degrees C increased thermotolerance at higher temperatures by approximately 1 degrees C. In parallel, the pretreatment increased Hsp70 expression suggesting a close link between Hsp70 expression and inducible thermotolerance. The increased Hsp70 levels after pretreatment were found to be due to the increased levels of the hsp70 RNA.


    Institute of Scientific and Technical Information of China (English)


    Objective To investigate the protective effects of heat shock protein 2700(Hsp27)on cis- platin inducing cytotoxicity in a temperature mutant SV40 large T antigen transformed rat embryo fibroblast (P1-Hsp27). Methods The cytotoxical effects of cisplatin on the proliferation status of P1-Hsp27 cells in the presence or absence of Hsp27 were measured by MTT assay. Results Cisplatin possessed dose-dependent cyto- toxicity on P1-Hsp27 cells. 48h after treatment, about 50% cells were dead in those cells exposed to 200μmol cisplatin. However, no obvious protective effects of Hsp27 on cisplatin induced cytotoxicity could be observed (P>0.05),except in those cells exposed to 500μmol cisplatin 12h after treatment. Conclusion Hsp27 has no obvious protective effects on cisplatin inducing cytotoxicity.

  4. Expression of HSP70 family and HSP90 family in skeletal muscles of transport stressed pigs%运输应激猪骨骼肌中热应激蛋白HSP70和HSP90家族的表达

    Institute of Scientific and Technical Information of China (English)

    鲍恩东; Sultan,KR; 等


    To demonstrate the expression of heat shock protein s(HSP)in skeletal muscles and to estimate the relationship between expression of HSP and pork quality, using Western blot four kinds of HSP which belongs to HSP70 family and HSP90 family respectively in porcine skeletal muscle tissues were determined after a long distance transport.All four HSP,namely HSP70,HSP72,HSP86 and HSP90,were regularly detected in M.longissimus dorsi and M.gluteus medius of both the transported and the cont rol group.The expression of HSP in control pigs showed that HSPs have some indep endent physiological from stress response undetermined functions.after 6 h trans port,the induction of the HSP70 and HSP72 in skeletal muscle tissues was slightly elevated(P>0.05).However,expression of the HSP 86 and the HSP90,especially HSP90,markedly decreased(P<0.01)in skelet al muscles.The results are implied that the reduction tendency of HSP90 fa mily is relative to pork quality and may serve as a potential marker for stress response.%利用Western blot技术,检测长途运输试验猪骨骼肌(背最长肌和臀大肌)中分属于HSP70家族和HSP90家族的4种应激蛋白(HSP70,HSP 72,HSP86和HSP90)的表达。所有运输应激猪和对照猪肌肉组织中均检测到了上述4种HSP。对照猪组织中HSP的表达说明HSP除了在受应激的细胞内行使生理功能外,还具有独立于应激刺激应答以外的其它作用。6 h的长途运输后,HSP的表达量明显不同,HSP70和HSP72在肌肉组织中的表达虽然有一定的增加,但统计学分析差异不显著(P>0.05);而HSP86和HSP90在骨骼肌中的表达明显下降,尤以HS P90下降最为显著(P<0.01)。提示HSP90家族成员与肉品品质相关,可能作为判断应激损伤的指征。

  5. Identification and in silico analysis of the Citrus HSP70 molecular chaperone gene family

    Directory of Open Access Journals (Sweden)

    Luciano G. Fietto


    Full Text Available The completion of the genome sequencing of the Arabidopsis thaliana model system provided a powerful molecular tool for comparative analysis of gene families present in the genome of economically relevant plant species. In this investigation, we used the sequences of the Arabidopsis Hsp70 gene family to identify and annotate the Citrus Hsp70 genes represented in the CitEST database. Based on sequence comparison analysis, we identified 18 clusters that were further divided into 5 subgroups encoding four mitochondrial mtHsp70s, three plastid csHsp70s, one ER luminal Hsp70 BiP, two HSP110/SSE-related proteins and eight cytosolic Hsp/Hsc70s. We also analyzed the expression profile by digital Northern of each Hsp70 transcript in different organs and in response to stress conditions. The EST database revealed a distinct population distribution of Hsp70 ESTs among isoforms and across the organs surveyed. The Hsp70-5 isoform was highly expressed in seeds, whereas BiP, mitochondrial and plastid HSp70 mRNAs displayed a similar expression profile in the organs analyzed, and were predominantly represented in flowers. Distinct Hsp70 mRNAs were also differentially expressed during Xylella infection and Citrus tristeza viral infection as well as during water deficit. This in silico study sets the groundwork for future investigations to fully characterize functionally the Citrus Hsp70 family and underscores the relevance of Hsp70s in response to abiotic and biotic stresses in Citrus.

  6. Genome-scale co-evolutionary inference identifies functions and clients of bacterial Hsp90.

    Directory of Open Access Journals (Sweden)

    Maximilian O Press

    Full Text Available The molecular chaperone Hsp90 is essential in eukaryotes, in which it facilitates the folding of developmental regulators and signal transduction proteins known as Hsp90 clients. In contrast, Hsp90 is not essential in bacteria, and a broad characterization of its molecular and organismal function is lacking. To enable such characterization, we used a genome-scale phylogenetic analysis to identify genes that co-evolve with bacterial Hsp90. We find that genes whose gain and loss were coordinated with Hsp90 throughout bacterial evolution tended to function in flagellar assembly, chemotaxis, and bacterial secretion, suggesting that Hsp90 may aid assembly of protein complexes. To add to the limited set of known bacterial Hsp90 clients, we further developed a statistical method to predict putative clients. We validated our predictions by demonstrating that the flagellar protein FliN and the chemotaxis kinase CheA behaved as Hsp90 clients in Escherichia coli, confirming the predicted role of Hsp90 in chemotaxis and flagellar assembly. Furthermore, normal Hsp90 function is important for wild-type motility and/or chemotaxis in E. coli. This novel function of bacterial Hsp90 agreed with our subsequent finding that Hsp90 is associated with a preference for multiple habitats and may therefore face a complex selection regime. Taken together, our results reveal previously unknown functions of bacterial Hsp90 and open avenues for future experimental exploration by implicating Hsp90 in the assembly of membrane protein complexes and adaptation to novel environments.

  7. Tumor imaging and targeting potential of an Hsp70-derived 14-mer peptide.

    Directory of Open Access Journals (Sweden)

    Mathias Gehrmann

    Full Text Available BACKGROUND: We have previously used a unique mouse monoclonal antibody cmHsp70.1 to demonstrate the selective presence of a membrane-bound form of Hsp70 (memHsp70 on a variety of leukemia cells and on single cell suspensions derived from solid tumors of different entities, but not on non-transformed cells or cells from corresponding 'healthy' tissue. This antibody can be used to image tumors in vivo and target them for antibody-dependent cellular cytotoxicity. Tumor-specific expression of memHsp70 therefore has the potential to be exploited for theranostic purposes. Given the advantages of peptides as imaging and targeting agents, this study assessed whether a 14-mer tumor penetrating peptide (TPP; TKDNNLLGRFELSG, the sequence of which is derived from the oligomerization domain of Hsp70 which is expressed on the cell surface of tumor cells, can also be used for targeting membrane Hsp70 positive (memHsp70+ tumor cells, in vitro. METHODOLOGY/PRINCIPAL FINDINGS: The specificity of carboxy-fluorescein (CF- labeled TPP (TPP to Hsp70 was proven in an Hsp70 knockout mammary tumor cell system. TPP specifically binds to different memHsp70+ mouse and human tumor cell lines and is rapidly taken up via endosomes. Two to four-fold higher levels of CF-labeled TPP were detected in MCF7 (82% memHsp70+ and MDA-MB-231 (75% memHsp70+ cells compared to T47D cells (29% memHsp70+ that exhibit a lower Hsp70 membrane positivity. After 90 min incubation, TPP co-localized with mitochondrial membranes in memHsp70+ tumors. Although there was no evidence that any given vesicle population was specifically localized, fluorophore-labeled cmHsp70.1 antibody and TPP preferentially accumulated in the proximity of the adherent surface of cultured cells. These findings suggest a potential association between membrane Hsp70 expression and cytoskeletal elements that are involved in adherence, the establishment of intercellular synapses and/or membrane reorganization. CONCLUSIONS

  8. Combination of Correctors Rescue ΔF508-CFTR by Reducing Its Association with Hsp40 and Hsp27. (United States)

    Lopes-Pacheco, Miquéias; Boinot, Clément; Sabirzhanova, Inna; Morales, Marcelo M; Guggino, William B; Cebotaru, Liudmila


    Correcting the processing of ΔF508-CFTR, the most common mutation in cystic fibrosis, is the major goal in the development of new therapies for this disease. Here, we determined whether ΔF508 could be rescued by a combination of small-molecule correctors, and identified the mechanism by which correctors rescue the trafficking mutant of cystic fibrosis transmembrane conductance regulator (CFTR). We transfected COS-7 cells with ΔF508, created HEK-293 stably expressing ΔF508, and utilized CFBE41o(-) cell lines stably transduced with ΔF508. As shown previously, ΔF508 expressed less protein, was unstable at physiological temperature, and rapidly degraded. When the cells were treated with the combination C18 + C4 the mature C-band was expressed at the cell surface. After treatment with C18 + C4, we saw a lower rate of protein disappearance after translation was stopped with cycloheximide. To understand how this rescue occurs, we evaluated the change in the binding of proteins involved in endoplasmic reticulum-associated degradation, such as Hsp27 (HspB1) and Hsp40 (DnaJ). We saw a dramatic reduction in binding to heat shock proteins 27 and 40 following combined corrector therapy. siRNA experiments confirmed that a reduction in Hsp27 or Hsp40 rescued CFTR in the ΔF508 mutant, but the rescue was not additive or synergistic with C4 + 18 treatment, indicating these correctors shared a common pathway for rescue involving a network of endoplasmic reticulum-associated degradation proteins.

  9. Heat Shock Protein HSP27 Secretion by Ovarian Cancer Cells Is Linked to Intracellular Expression Levels, Occurs Independently of the Endoplasmic Reticulum Pathway and HSP27’s Phosphorylation Status, and Is Mediated by Exosome Liberation

    Directory of Open Access Journals (Sweden)

    Matthias B. Stope


    Full Text Available The heat shock protein HSP27 has been correlated in ovarian cancer (OC patients with aggressiveness and chemoresistance and, therefore, represents a promising potential biomarker for OC diagnosis, prognosis, and treatment response. Notably, secretion of soluble HSP27 has been described by a few cell types and may take place as well in OC cells. Therefore, we studied HSP27 secretion mechanisms under diverse cellular conditions in an OC cell model system. Secretion of HSP27 was characterized after overexpression of HSP27 by transfected plasmids and after heat shock. Intra- and extracellular HSP27 amounts were assessed by Western blotting and ELISA. Protein secretion was blocked by brefeldin A and the impact of the HSP27 phosphorylation status was analyzed overexpressing HSP27 phosphomutants. The present study demonstrated that HSP27 secretion by OVCAR-3 and SK-OV-3 cells depends on intracellular HSP27 concentrations. Moreover, HSP27 secretion is independent of the endoplasmic reticulum secretory pathway and HSP27 phosphorylation. Notably, analysis of OC cell-born exosomes not only confirmed the concentration-dependent correlation of HSP27 expression and secretion but also demonstrated a concentration-dependent incorporation of HSP27 protein into exosomes. Thus, secreted HSP27 may become more important as an extracellular factor which controls the tumor microenvironment and might be a noninvasive biomarker.

  10. Cloning and Expression of a Cytosolic HSP90 Gene in Chlorella vulgaris

    Directory of Open Access Journals (Sweden)

    Zhengyi Liu


    Full Text Available Heat shock protein 90 (HSP90, a highly conserved molecular chaperone, plays essential roles in folding, keeping structural integrity, and regulating the subset of cytosolic proteins. We cloned the cDNA of Chlorella vulgaris HSP90 (named CvHSP90 by combining homology cloning with rapid amplification of cDNA ends (RACE. Sequence analysis indicated that CvHSP90 is a cytosolic member of the HSP90 family. Quantitative RT-PCR was applied to determine the expression level of messenger RNA (mRNA in CvHSP90 under different stress conditions. C. vulgaris was kept in different temperatures (5–45°C for 1 h. The mRNA expression level of CvHSP90 increased with temperature from 5 to 10°C, went further from 35 to 40°C, and reached the maximum at 40°C. On the other hand, for C. vulgaris kept at 35°C for different durations, the mRNA expression level of CvHSP90 increased gradually and reached the peak at 7 h and then declined progressively. In addition, the expression level of CvHSP90 at 40 or 45 in salinity (‰ was almost fourfold of that at 25 in salinity (‰ for 2 h. Therefore, CvHSP90 may be a potential biomarker to monitor environment changes.

  11. Preferential secretion of inducible HSP70 by vitiligo melanocytes under stress. (United States)

    Mosenson, Jeffrey A; Flood, Kelsey; Klarquist, Jared; Eby, Jonathan M; Koshoffer, Amy; Boissy, Raymond E; Overbeck, Andreas; Tung, Rebecca C; Le Poole, I Caroline


    Inducible HSP70 (HSP70i) chaperones peptides from stressed cells, protecting them from apoptosis. Upon extracellular release, HSP70i serves an adjuvant function, enhancing immune responses to bound peptides. We questioned whether HSP70i differentially protects control and vitiligo melanocytes from stress and subsequent immune responses. We compared expression of HSP70i in skin samples, evaluated the viability of primary vitiligo and control melanocytes exposed to bleaching phenols, and measured secreted HSP70i. We determined whether HSP70i traffics to melanosomes to contact immunogenic proteins by cell fractionation, western blotting, electron microscopy, and confocal microscopy. Viability of vitiligo and control melanocytes was equally affected under stress. However, vitiligo melanocytes secreted increased amounts of HSP70i in response to MBEH, corroborating with aberrant HSP70i expression in patient skin. Intracellular HSP70i colocalized with melanosomes, and more so in response to MBEH in vitiligo melanocytes. Thus, whereas either agent is cytotoxic to melanocytes, MBEH preferentially induces immune responses to melanocytes.

  12. Calcium Homeostasis and Muscle Energy Metabolism Are Modified in HspB1-Null Mice (United States)

    Picard, Brigitte; Kammoun, Malek; Gagaoua, Mohammed; Barboiron, Christiane; Meunier, Bruno; Chambon, Christophe; Cassar-Malek, Isabelle


    Hsp27—encoded by HspB1—is a member of the small heat shock proteins (sHsp, 12–43 kDa (kilodalton)) family. This protein is constitutively present in a wide variety of tissues and in many cell lines. The abundance of Hsp27 is highest in skeletal muscle, indicating a crucial role for muscle physiology. The protein identified as a beef tenderness biomarker was found at a crucial hub in a functional network involved in beef tenderness. The aim of this study was to analyze the proteins impacted by the targeted invalidation of HspB1 in the Tibialis anterior muscle of the mouse. Comparative proteomics using two-dimensional gel electrophoresis revealed 22 spots that were differentially abundant between HspB1-null mice and their controls that could be identified by mass spectrometry. Eighteen spots were more abundant in the muscle of the mutant mice, and four were less abundant. The proteins impacted by the absence of Hsp27 belonged mainly to calcium homeostasis (Srl and Calsq1), contraction (TnnT3), energy metabolism (Tpi1, Mdh1, PdhB, Ckm, Pygm, ApoA1) and the Hsp proteins family (HspA9). These data suggest a crucial role for these proteins in meat tenderization. The information gained by this study could also be helpful to predict the side effects of Hsp27 depletion in muscle development and pathologies linked to small Hsps.

  13. A novel biomarker for marine environmental pollution of HSP90 from Mytilus coruscus. (United States)

    Liu, Huihui; Wu, Jiong; Xu, Mengshan; He, Jianyu


    Heat shock protein 90 (HSP90) is a conserved molecular chaperone contributing to cell cycle control, organism development and the proper regulation of cytosolic proteins. The full-length HSP90 cDNA of Mytilus coruscus (McHSP90, KT946644) was 2420bp, including an ORF of 2169bp encoding a polypeptide of 722 amino acids with predicted pI/MW 4.89/83.22kDa. BLASTp analysis and phylogenetic relationship strongly suggested McHSP90 was a member of HSP90 family, and it was highly conserved with other known HSP90, especially in the HSP90 family signatures, ATP/GTP-Binding sites and 'EEVD' motif. The mRNA of McHSP90 in haemolymph was upregulated in all treatments including Vibrio alginolyticus and Vibrio harveyi challenge, metals stresses (copper and cadmium) and 180 CST fuel exposure. All the results implied the expression of McHSP90 could be affected by Vibrio challenge and environmental stress, which might help us gain more insight into the molecular mechanism of HSP against adverse stresses in mollusca.

  14. Studies on effect of different seasons on expression of HSP70 and HSP90 gene in sperm of Tharparkar bull semen

    Directory of Open Access Journals (Sweden)

    J.S. Rajoriya


    Conclusion: It was concluded from the present study that there was no significant difference in the mRNA expression of HSP 70 and HSP 90 between the winter and summer season, presence of similar type of stress resistant spermatozoa in Tharparkar bull semen and the semen can be cryopreserved throughout the year in this prestigious Indian breed.

  15. Effect of vibrational stress and spaceflight on regulation of heat shock proteins hsp70 and hsp27 in human lymphocytes (Jurkat) (United States)

    Cubano, L. A.; Lewis, M. L.


    Heat shock protein levels are increased in cells as a result of exposure to stress. To determine whether heat shock protein regulation could be used to evaluate stress in cells during spaceflight, the response of Jurkat cells to spaceflight and simulated space shuttle launch vibration was investigated by evaluating hsp70 and hsp27 gene expression. Gene expression was assessed by reverse transcription-polymerase chain reaction using mRNA extracted from vibrated, nonvibrated, space-flown, and ground control cells. Results indicate that mechanical stresses of vibration and low gravity do not up-regulate the mRNA for hsp70, although the gene encoding hsp27 is up-regulated by spaceflight but not by vibration. In ground controls, the mRNA for hsp70 and hsp27 increased with time in culture. We conclude that hsp70 gene expression is a useful indicator of stress related to culture density but is not an indicator of the stresses of launch vibration or microgravity. Up-regulation of hsp27 gene expression in microgravity is a new finding.

  16. Functional Divergence of Hsp90 Genetic Interactions in Biofilm and Planktonic Cellular States.

    Directory of Open Access Journals (Sweden)

    Stephanie Diezmann

    Full Text Available Candida albicans is among the most prevalent opportunistic fungal pathogens. Its capacity to cause life-threatening bloodstream infections is associated with the ability to form biofilms, which are intrinsically drug resistant reservoirs for dispersal. A key regulator of biofilm drug resistance and dispersal is the molecular chaperone Hsp90, which stabilizes many signal transducers. We previously identified 226 C. albicans Hsp90 genetic interactors under planktonic conditions, of which 56 are involved in transcriptional regulation. Six of these transcriptional regulators have previously been implicated in biofilm formation, suggesting that Hsp90 genetic interactions identified in planktonic conditions may have functional significance in biofilms. Here, we explored the relationship between Hsp90 and five of these transcription factor genetic interactors: BCR1, MIG1, TEC1, TUP1, and UPC2. We deleted each transcription factor gene in an Hsp90 conditional expression strain, and assessed biofilm formation and morphogenesis. Strikingly, depletion of Hsp90 conferred no additional biofilm defect in the mutants. An interaction was observed in which deletion of BCR1 enhanced filamentation upon reduction of Hsp90 levels. Further, although Hsp90 modulates expression of TEC1, TUP1, and UPC2 in planktonic conditions, it has no impact in biofilms. Lastly, we probed for physical interactions between Hsp90 and Tup1, whose WD40 domain suggests that it might interact with Hsp90 directly. Hsp90 and Tup1 formed a stable complex, independent of temperature or developmental state. Our results illuminate a physical interaction between Hsp90 and a key transcriptional regulator of filamentation and biofilm formation, and suggest that Hsp90 has distinct genetic interactions in planktonic and biofilm cellular states.

  17. Hsp90 is a direct target of the anti-allergic drugs disodium cromoglycate and amlexanox. (United States)

    Okada, Miki; Itoh, Hideaki; Hatakeyama, Takashi; Tokumitsu, Hiroshi; Kobayashi, Ryoji


    Hsp90 (heat-shock protein 90) alone can act to prevent protein aggregation and promote refolding in vitro, but in vivo it operates as a part of a multichaperone complex, which includes Hsp70 and cohort proteins. Since the physiological function of Hsp90 is not yet fully understood, the development of specific antagonists might open new lines of investigation on the role of Hsp90. In an effort to discover Hsp90 antagonists, we screened many drugs and found that the anti-allergic drugs DSCG (disodium cromoglycate) and amlexanox target Hsp90. Both drugs were found to bind directly wild-type Hsp90 via the N- and C-terminal domains. Both drugs strongly suppressed the in vitro chaperone activity of native Hsp90 towards citrate synthase at 1.5-3.0 microM. Amlexanox suppressed C-terminal chaperone activity in vitro, but not N-terminal chaperone activity, and inhibited the association of cohort proteins, such as cyclophilin 40 and Hsp-organizing protein, to the C-terminal domain of Hsp90. These data suggest that amlexanox might disrupt the multichaperone complex, including Hsp70 and cohort proteins, both in vitro and in vivo. Although DSCG inhibited the in vitro chaperone activity of the N-terminal domain, the drug had no effect either on the C-terminal chaperone activity or on the association of the cohort proteins with the C-terminus of Hsp90. The physiological significance of these interactions in vivo remains to be investigated further, but undoubtedly must be taken into account when considering the pharmacology of anti-allergic drugs. DSCG and amlexanox may serve as useful tools for evaluating the physiological significance of Hsp90.

  18. Hsp90 governs echinocandin resistance in the pathogenic yeast Candida albicans via calcineurin.

    Directory of Open Access Journals (Sweden)

    Sheena D Singh


    Full Text Available Candida albicans is the leading fungal pathogen of humans, causing life-threatening disease in immunocompromised individuals. Treatment of candidiasis is hampered by the limited number of antifungal drugs whose efficacy is compromised by host toxicity, fungistatic activity, and the emergence of drug resistance. We previously established that the molecular chaperone Hsp90, which regulates the form and function of diverse client proteins, potentiates resistance to the azoles in C. albicans and in the model yeast Saccharomyces cerevisiae. Genetic studies in S. cerevisiae revealed that Hsp90's role in azole resistance is to enable crucial cellular responses to the membrane stress exerted by azoles via the client protein calcineurin. Here, we demonstrate that Hsp90 governs cellular circuitry required for resistance to the only new class of antifungals to reach the clinic in decades, the echinocandins, which inhibit biosynthesis of a critical component of the fungal cell wall. Pharmacological or genetic impairment of Hsp90 function reduced tolerance of C. albicans laboratory strains and resistance of clinical isolates to the echinocandins and created a fungicidal combination. Compromising calcineurin function phenocopied compromising Hsp90 function. We established that calcineurin is an Hsp90 client protein in C. albicans: reciprocal co-immunoprecipitation validated physical interaction; Hsp90 inhibition blocked calcineurin activation; and calcineurin levels were depleted upon genetic reduction of Hsp90. The downstream effector of calcineurin, Crz1, played a partial role in mediating calcineurin-dependent stress responses activated by echinocandins. Hsp90's role in echinocandin resistance has therapeutic potential given that genetic compromise of C. albicans HSP90 expression enhanced the efficacy of an echinocandin in a murine model of disseminated candidiasis. Our results identify the first Hsp90 client protein in C. albicans, establish an entirely

  19. Cell stress promotes the association of phosphorylated HspB1 with F-actin.

    Directory of Open Access Journals (Sweden)

    Joseph P Clarke

    Full Text Available Previous studies have suggested that the small heat shock protein, HspB1, has a direct influence on the dynamics of cytoskeletal elements, in particular, filamentous actin (F-actin polymerization. In this study we have assessed the influence of HspB1 phosphorylation on its interaction(s with F-actin. We first determined the distribution of endogenous non-phosphorylated HspB1, phosphorylated HspB1 and F-actin in neuroendocrine PC12 cells by immunocytochemistry and confocal microscopy. We then investigated a potential direct interaction between HspB1 with F-actin by precipitating F-actin directly with biotinylated phalloidin followed by Western analyses; the reverse immunoprecipitation of HspB1 was also carried out. The phosphorylation influence of HspB1 in this interaction was investigated by using pharmacologic inhibition of p38 MAPK. In control cells, HspB1 interacts with F-actin as a predominantly non-phosphorylated protein, but subsequent to stress there is a redistribution of HspB1 to the cytoskeletal fraction and a significantly increased association of pHspB1 with F-actin. Our data demonstrate HspB1 is found in a complex with F-actin both in phosphorylated and non-phosphorylated forms, with an increased association of pHspB1 with F-actin after heat stress. Overall, our study combines both cellular and biochemical approaches to show cellular localization and direct demonstration of an interaction between endogenous HspB1 and F-actin using methodolgy that specifically isolates F-actin.

  20. Changes in HSP gene and protein expression in natural scrapie with brain damage

    Directory of Open Access Journals (Sweden)

    Serrano Carmen


    Full Text Available Abstract Heat shock proteins (Hsp perform cytoprotective functions such as apoptosis regulation and inflammatory response control. These proteins can also be secreted to the extracellular medium, acting as inflammatory mediators, and their chaperone activity permits correct folding of proteins and avoids the aggregation of anomalous isoforms. Several studies have proposed the implication of Hsp in prion diseases. We analysed the gene expression and protein distribution of different members of the Hsp27, Hsp70, and Hsp90 families in the central nervous system of sheep naturally infected with scrapie. Different expression profiles were observed in the areas analysed. Whereas changes in transcript levels were not observed in the cerebellum or medulla oblongata, a significant decrease in HSP27 and HSP90 was detected in the prefrontal cortex. In contrast, HSP73 was over-expressed in diencephalons of scrapie animals. Western blotting did not reveal significant differences in Hsp90 and Hsp70 protein expression between scrapie and control animals. Expression rates identified by real-time RT-PCR and western blotting were compared with the extent of classical scrapie lesions using stepwise regression. Changes in Hsp gene and protein expression were associated with prion protein deposition, gliosis and spongiosis rather than with apoptosis. Finally, immunohistochemistry revealed intense Hsp70 and Hsp90 immunolabelling in Purkinje cells of scrapie sheep. In contrast, controls displayed little or no staining in these cells. The observed differences in gene expression and protein distribution suggest that the heat shock proteins analysed play a role in the natural form of the disease.

  1. A Cytosolic Relay of Heat Shock Proteins HSP70-1A and HSP90β Monitors the Folding Trajectory of the Serotonin Transporter* (United States)

    El-Kasaby, Ali; Koban, Florian; Sitte, Harald H.; Freissmuth, Michael; Sucic, Sonja


    Mutations in the C terminus of the serotonin transporter (SERT) disrupt folding and export from the endoplasmic reticulum. Here we examined the hypothesis that a cytosolic heat shock protein relay was recruited to the C terminus to assist folding of SERT. This conjecture was verified by the following observations. (i) The proximal portion of the SERT C terminus conforms to a canonical binding site for DnaK/heat shock protein of 70 kDa (HSP70). A peptide covering this segment stimulated ATPase activity of purified HSP70-1A. (ii) A GST fusion protein comprising the C terminus of SERT pulled down HSP70-1A. The interaction between HSP70-1A and SERT was visualized in live cells by Förster resonance energy transfer: it was restricted to endoplasmic reticulum-resident transporters and enhanced by an inhibitor that traps HSP70-1A in its closed state. (iv) Co-immunoprecipitation confirmed complex formation of SERT with HSP70-1A and HSP90β. Consistent with an HSP relay, co-chaperones (e.g. HSC70-HSP90-organizing protein) were co-immunoprecipitated with the stalled mutants SERT-R607A/I608A and SERT-P601A/G602A. (v) Depletion of HSP90β by siRNA or its inhibition increased the cell surface expression of wild type SERT and SERT-F604Q. In contrast, SERT-R607A/I608A and SERT-P601A/G602A were only rendered susceptible to inhibition of HSP70 and HSP90 by concomitant pharmacochaperoning with noribogaine. (vi) In JAR cells, inhibition of HSP90 also increased the levels of SERT, indicating that endogenously expressed transporter was also susceptible to control by HSP90β. These findings support the concept that the folding trajectory of SERT is sampled by a cytoplasmic chaperone relay. PMID:25202009

  2. Low sequence identity but high structural and functional conservation: The case of Hsp70/Hsp90 organizing protein (Hop/Sti1) of Leishmania braziliensis. (United States)

    Batista, Fernanda A H; Seraphim, Thiago V; Santos, Clelton A; Gonzaga, Marisvanda R; Barbosa, Leandro R S; Ramos, Carlos H I; Borges, Júlio C


    Parasites belonging to the genus Leishmania are subjected to extensive environmental changes during their life cycle; molecular chaperones/co-chaperones act as protagonists in this scenario to maintain cellular homeostasis. Hop/Sti1 is a co-chaperone that connects the Hsp90 and Hsp70 systems, modulating their ATPase activities and affecting the fate of client proteins because it facilitates their transfer from the Hsp70 to the Hsp90 chaperone. Hop/Sti1 is one of the most prevalent co-chaperones, highlighting its importance despite the relatively low sequence identity among orthologue proteins. This multi-domain protein comprises three tetratricopeptides domains (TPR1, TPR2A and TPR2B) and two Asp/Pro-rich domains. Given the importance of Hop/Sti1 for the chaperone system and for Leishmania protozoa viability, the Leishmania braziliensis Hop (LbHop) and a truncated mutant (LbHop(TPR2AB)) were characterized. Structurally, both proteins are α-helix-rich and highly elongated monomeric proteins. Functionally, they inhibited the ATPase activity of Leishmania braziliensis Hsp90 (LbHsp90) to a similar extent, and the thermodynamic parameters of their interactions with LbHsp90 were similar, indicating that TPR2A-TPR2B forms the functional center for the LbHop interaction with LbHsp90. These results highlight the structural and functional similarity of Hop/Sti1 proteins, despite their low sequence conservation compared to the Hsp70 and Hsp90 systems, which are phylogenetic highly conserved.

  3. Gene expression profiles of cytosolic heat shock proteins Hsp70 and Hsp90 from symbiotic dinoflagellates in response to thermal stress: possible implications for coral bleaching. (United States)

    Rosic, Nedeljka N; Pernice, Mathieu; Dove, Sophie; Dunn, Simon; Hoegh-Guldberg, Ove


    Unicellular photosynthetic dinoflagellates of the genus Symbiodinium are the most common endosymbionts of reef-building scleractinian corals, living in a symbiotic partnership known to be highly susceptible to environmental changes such as hyperthermic stress. In this study, we identified members of two major heat shock proteins (HSPs) families, Hsp70 and Hsp90, in Symbiodinium sp. (clade C) with full-length sequences that showed the highest similarity and evolutionary relationship with other known HSPs from dinoflagellate protists. Regulation of HSPs gene expression was examined in samples of the scleractinian coral Acropora millepora subjected to elevated temperatures progressively over 18 h (fast) and 120 h (gradual thermal stress). Moderate to severe heat stress at 26°C and 29°C (+3°C and +6°C above average sea temperature) resulted in an increase in algal Hsp70 gene expression from 39% to 57%, while extreme heat stress (+9°C) reduced Hsp70 transcript abundance by 60% (after 18 h) and 70% (after 120 h). Elevated temperatures decreased an Hsp90 expression under both rapid and gradual heat stress scenarios. Comparable Hsp70 and Hsp90 gene expression patterns were observed in Symbiodinium cultures and in hospite, indicating their independent regulation from the host. Differential gene expression profiles observed for Hsp70 and Hsp90 suggests diverse roles of these molecular chaperones during heat stress response. Reduced expression of the Hsp90 gene under heat stress can indicate a reduced role in inhibiting the heat shock transcription factor which may lead to activation of heat-inducible genes and heat acclimation.

  4. 草鱼hsp70和hsp90对温度急性变化的响应%Effect of rapid temperature change on expression of hsp70 and hsp90 in grass carp(Ctenopharyngodon idella)

    Institute of Scientific and Technical Information of China (English)

    周鑫; 董云伟; 王芳; 董双林


    Grass carp ( Ctenopharyngodon idella) is one of the most important cultured fishes in China. During transportation in summer,massive mortality often occurs. As molecular chaperones,Hsps assist cells in their recovery from stress and promote cytoprotection. Limited reports described the expression of Hsps in grass carp at high temperatures, and the relationship between oxygen consumption, expression of heat shock protein and mortality needs to be investigated based on the oxygen- and capability- limited thermal tolerance theory (OCLTT) to further elucidate the thermal adaptation of this species to high temperature in view of integrated biology. In the present study,the expressions of hsp70 and hsp90 after rapid temperature increase were studied. Grass carps were acclimated at 20 t and then exposed to designated temperatures(20,22,24, 26,28,30,32,34 ℃) for 3 h heat shock. After 2 h-recovery at 20℃, liver, muscle and gill of tested fish were sampled and hsp70 and hsp90 were determined using semi-quantitive real-time PCR. Expression of hsps increased with temperature increase until 32℃ in muscle and gill, and then decreased at 34 ℃. In liver, expression of hsps kept increasing in all temperature treatments. These results indicated that hsp70 and hsp90 were sensitive to temperature increase in grass carp. Analyzed heat shock proteins data with previous results of oxygen consumption and lethal temperatures of grass carp, the physiological adaptations of grass carp could be explained by oxygen- and capability- limited thermal tolerance theory (OCLTT), which pointed out that the imbalance between oxygen demand and tissue oxygen supply ability was the primary factor which limited organisms' thermal tolerance and the ability of aerobic metabolism greatly limited the survival of organism beyond critical temperatures. With temperature increase, the maximum value of oxygen consumption in grass carp occurred at 28 ℃, and then the oxygen consumption decreased when

  5. Cytochemical identification of HSP110 during early mouse facial development. (United States)

    Evrard, L; Vanmuylder, N; Dourov, N; Glineur, R; Louryan, S


    Apoptotic cell death constitutes a common phenomenon observed during development. This process plays an important role in the regulation of cell populations and in early differentiation of embryonic organs. Several teratologic situations are considered as resulting in a dramatic increase of the apoptotic process. In mammalian cells, heat shock proteins (HSPs), expressed or increased in response to various stresses, act as molecular chaperones in physiological conditions. In order to determine specific histochemical markers of apoptotic cells in normal craniofacial development, we observed the expression of stress proteins (HSPs) 70, 86, and 110. The apoptotic pattern of mesectodermal cell death areas was confirmed using both nuclear staining (Feulgen) and specific labeling of DNA fragmentation (TUNEL). These areas are localized in the proximal parts of the first and second visceral arches. They are located in mesectodermal and ganglionic cells. Apoptotic mesectodermal populations strongly express HSP110, as shown by the cytochemical identification of HSP110 and by double staining HSP110-TUNEL, suggesting that this protein could be considered as a new marker for apoptotic embryonic cells, and could be used in further teratologic studies to better quantify induced cell death.

  6. The HSP90 Inhibitor Ganetespib Radiosensitizes Human Lung Adenocarcinoma Cells

    Energy Technology Data Exchange (ETDEWEB)

    Gomez-Casal, Roberto; Bhattacharya, Chitralekha [The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213 (United States); Department of Medicine, The University of Pittsburgh, Pittsburgh, PA 15213 (United States); Epperly, Michael W. [The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213 (United States); Department of Radiation Oncology, The University of Pittsburgh, Pittsburgh, PA 15213 (United States); Basse, Per H. [The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213 (United States); Department of Immunology, The University of Pittsburgh, Pittsburgh, PA 15213 (United States); Wang, Hong [The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213 (United States); Department of Biostatistics, The University of Pittsburgh, Pittsburgh, PA 15213 (United States); Wang, Xinhui [Harvard Medical School, Harvard University, 25 Shattuck Street, Boston, MA 02115 (United States); Proia, David A. [Synta Pharmaceuticals Corp., 45 Hartwell Avenue, Lexington, MA 02421 (United States); Greenberger, Joel S. [The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213 (United States); Department of Radiation Oncology, The University of Pittsburgh, Pittsburgh, PA 15213 (United States); Socinski, Mark A.; Levina, Vera, E-mail: [The University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213 (United States); Department of Medicine, The University of Pittsburgh, Pittsburgh, PA 15213 (United States)


    The molecular chaperone HSP90 is involved in stabilization and function of multiple client proteins, many of which represent important oncogenic drivers in NSCLC. Utilization of HSP90 inhibitors as radiosensitizing agents is a promising approach. The antitumor activity of ganetespib, HSP90 inhibitor, was evaluated in human lung adenocarcinoma (AC) cells for its ability to potentiate the effects of IR treatment in both in vitro and in vivo. The cytotoxic effects of ganetespib included; G2/M cell cycle arrest, inhibition of DNA repair, apoptosis induction, and promotion of senescence. All of these antitumor effects were both concentration- and time-dependent. Both pretreatment and post-radiation treatment with ganetespib at low nanomolar concentrations induced radiosensitization in lung AC cells in vitro. Ganetespib may impart radiosensitization through multiple mechanisms: such as down regulation of the PI3K/Akt pathway; diminished DNA repair capacity and promotion of cellular senescence. In vivo, ganetespib reduced growth of T2821 tumor xenografts in mice and sensitized tumors to IR. Tumor irradiation led to dramatic upregulation of β-catenin expression in tumor tissues, an effect that was mitigated in T2821 xenografts when ganetespib was combined with IR treatments. These data highlight the promise of combining ganetespib with IR therapies in the treatment of AC lung tumors.

  7. Identification of an Allosteric Pocket on Human Hsp70 Reveals a Mode of Inhibition of This Therapeutically Important Protein (United States)

    Rodina, Anna; Patel, Pallav D.; Kang, Yanlong; Patel, Yogita; Baaklini, Imad; Wong, Michael J.H.; Taldone, Tony; Yan, Pengrong; Yang, Chenghua; Maharaj, Ronnie; Gozman, Alexander; Patel, Maulik R.; Patel, Hardik J.; Chirico, William; Erdjument-Bromage, Hediye; Talele, Tanaji T.; Young, Jason C.; Chiosis, Gabriela


    SUMMARY Hsp70s are important cancer chaperones that act upstream of Hsp90 and exhibit independent anti-apoptotic activities. To develop chemical tools for the study of human Hsp70, we developed a homology model that unveils a previously unknown allosteric site located in the nucleotide binding domain of Hsp70. Combining structure-based design and phenotypic testing, we discovered a previously unknown inhibitor of this site, YK5. In cancer cells, this compound is a potent and selective binder of the cytosolic but not the organellar human Hsp70s and has biological activity partly by interfering with the formation of active oncogenic Hsp70/Hsp90/client protein complexes. YK5 is a small molecule inhibitor rationally designed to interact with an allosteric pocket of Hsp70 and represents a previously unknown chemical tool to investigate cellular mechanisms associated with Hsp70. PMID:24239008

  8. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard


    We show that inhibition of HDAC activity leads to surface expression of Hsp70 on various hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC inhibitor-mediated Hsp70 surface expression was confined to the apoptotic Annexin V...... activity selectively induces surface expression of Hsp70 on hematopoietic cancer cells and that this may increase immunorecognition of these cells.......-positive cells and blocked by inhibition of apoptosis. Other chemotherapeutic inducers of apoptosis such as etoposide and camptothecin also led to a robust induction of Hsp70 surface expression. Hsp70 expression was, however, not caused by induction of apoptosis per se, as activated CD4 T cells remained Hsp70...

  9. Clinical application of bio ceramics (United States)

    Anu, Sharma; Gayatri, Sharma


    Ceramics are the inorganic crystalline material. These are used in various field such as biomedical, electrical, electronics, aerospace, automotive and optical etc. Bio ceramics are the one of the most active areas of research. Bio ceramics are the ceramics which are biocompatible. The unique properties of bio ceramics make them an attractive option for medical applications and offer some potential advantages over other materials. During the past three decades, a number of major advances have been made in the field of bio ceramics. This review focuses on the use of these materials in variety of clinical scenarios.

  10. bioética

    Directory of Open Access Journals (Sweden)

    Francisco Javier León Correa


    Full Text Available Desde el inicio de la publicación de la revista Cuadernos de Bioética, hemos podido comprobar una evolución importante en el desarrollo de esta disciplina, tanto en España como en Latinoamérica, que son los ámbitos donde muy especialmente se difunde. Trataré de señalar los aspectos que considero de mayor interés en los inicios y desarrollo de esta nueva disciplina, para poder entender mejor los retos actualesque enfrenta y cuáles son las líneas que ha seguido la revista en esta andadura inicial, y que deberá seguir para responder a los retos de los próximos años. Es un reto la realidad que vivimos en nuestros países, con la aprobación de leyes impuestas desde los poderes públicos con muy poca o ninguna discusión social que se parezca de lejos al diálogo tolerante racional que propugna la Bioética; con las reformas aceleradas e indecisas de los sistemas de salud; con el cambio del rol del médico y de la relación médico-paciente desde un paternalismo aún presente a un modelo participativo o contractual sin defi nir claramente; con importantes desigualdades en el acceso a la atención de salud dentro de cada propio país, pero a la vez con un creciente reconocimiento de los derechos de los pacientes y de su necesaria participación para mejorar la calidad de la asistencia clínica. Tiene especial importancia valorar cuáles han sido los instrumentos del éxito y de la difusión de la Bioética clínica, y cuáles pueden ser las nuevas propuestas para el desarrollo de la Bioética como una disciplina de debate ético social y político, ligada a las exigencias éticas de la justicia y el desarrollo social. Hemos de ver cuál ha sido el aporte en estos quince años de la revista Cuadernos de Bioética, con la colaboración de cientos de autores de diversos países, en que se ha mantenido fi el a la propuesta de los que la iniciamos en Santiago de Compostela en el año 1990: construir un buen aporte al debate actual de Bio

  11. Chlamydia trachomatis infection and anti-Hsp60 immunity: the two sides of the coin.

    Directory of Open Access Journals (Sweden)

    Francesco Cappello


    Full Text Available Chlamydia trachomatis (CT infection is one of the most common causes of reproductive tract diseases and infertility. CT-Hsp60 is synthesized during infection and is released in the bloodstream. As a consequence, immune cells will produce anti-CT-Hsp60 antibodies. Hsp60, a ubiquitous and evolutionarily conserved chaperonin, is normally sequestered inside the cell, particularly into mitochondria. However, upon cell stress, as well as during carcinogenesis, the chaperonin becomes exposed on the cell surface (sf-Hsp60 and/or is secreted from cells into the extracellular space and circulation. Reports in the literature on circulating Hsp and anti-Hsp antibodies are in many cases short on details about Hsp60 concentrations, and about the specificity spectra of the antibodies, their titers, and their true, direct, pathogenetic effects. Thus, more studies are still needed to obtain a definitive picture on these matters. Nevertheless, the information already available indicates that the concurrence of persistent CT infection and appearance of sf-Hsp60 can promote an autoimmune aggression towards stressed cells and the development of diseases such as autoimmune arthritis, multiple sclerosis, atherosclerosis, vasculitis, diabetes, and thyroiditis, among others. At the same time, immunocomplexes composed of anti-CT-Hsp60 antibodies and circulating Hsp60 (both CT and human may form deposits in several anatomical locations, e.g., at the glomerular basal membrane. The opposite side of the coin is that pre-tumor and tumor cells with sf-Hsp60 can be destroyed with participation of the anti-Hsp60 antibody, thus stopping cancer progression before it is even noticed by the patient or physician.

  12. Hsp72 overexpression protects from APAP and MCD induced liver injury via attenuation of JNK signalling


    Levada, Kateryna


    Hsp72 is a classic, stress-inducible heat shock protein. It protects the organism from variety of diseases and stress situations. Because of its established cytoprotective function and stress-inducible expression Hsp72 plays an important role in different human disorders. However, its hepatic function remains largely unknown due to a lack of a suitable transgenic model. To study the hepatic function of Hsp72, I analyzed its expression in patients with liver disease and in newly generated tran...

  13. Recovery From Radiation-induced Bone Marrow Damage by HSP25 Through Tie2 Signaling

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hae-June [Division of Radiation Effects, Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of); Kwon, Hee-Chung [Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of); Chung, Hee-Yong [College of Medicine, Hanyang University, Seoul (Korea, Republic of); Lee, Yoon-Jin [Division of Radiation Effects, Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of); Lee, Yun-Sil, E-mail: [College of Pharmacy and Division of Life and Pharmaceutical Sciences, Ewha Woman' s University, Seoul (Korea, Republic of)


    Purpose: Whole-body radiation therapy can cause severe injury to the hematopoietic system, and therefore it is necessary to identify a novel strategy for overcoming this injury. Methods and Materials: Mice were irradiated with 4.5 Gy after heat shock protein 25 (HSP25) gene transfer using an adenoviral vector. Then, peripheral blood cell counts, histopathological analysis, and Western blotting on bone marrow (BM) cells were performed. The interaction of HSP25 with Tie2 was investigated with mouse OP9 and human BM-derived mesenchymal stem cells to determine the mechanism of HSP25 in the hematopoietic system. Results: HSP25 transfer increased BM regeneration and reduced apoptosis following whole-body exposure to ionizing radiation (IR). The decrease in Tie2 protein expression that followed irradiation of the BM was blocked by HSP25 transfer, and Tie2-positive cells were more abundant among the BM cells of HSP25-transferred mice, even after IR exposure. Following systemic RNA interference of Tie2 before IR, HSP25-mediated radioprotective effects were partially blocked in both mice and cell line systems. Stability of Tie2 was increased by HSP25, a response mediated by the interaction of HSP25 with Tie2. IR-induced tyrosine phosphorylation of Tie2 was augmented by HSP25 overexpression; downstream events in the Tie2 signaling pathway, including phosphorylation of AKT and EKR1/2, were also activated. Conclusions: HSP25 protects against radiation-induced BM damage by interacting with and stabilizing Tie2. This may be a novel strategy for HSP25-mediated radioprotection in BM.

  14. A bichaperone (Hsp70-Hsp78) system restores mitochondrial DNA synthesis following thermal inactivation of Mip1p polymerase. (United States)

    Germaniuk, Aleksandra; Liberek, Krzysztof; Marszalek, Jaroslaw


    Mitochondrial DNA synthesis is a thermosensitive process in the yeast Saccharomyces cerevisiae. We found that restoration of mtDNA synthesis following heat treatment of cells is dependent on reactivation of the mtDNA polymerase Mip1p through the action of a mitochondrial bichaperone system consisting of the Hsp70 system and the Hsp78 oligomeric protein. mtDNA synthesis was inefficiently restored after heat shock in yeast lacking either functional component of the bichaperone system. Furthermore, the activity of purified Mip1p was also thermosensitive; however, the purified components of the mitochondrial bichaperone system (Ssc1p, Mdj1p, Mge1p, and Hsp78p) were able to protect its activity under moderate heat shock conditions as well as to reactivate thermally inactivated Mip1p. Interestingly, the reactivation of endogenous Mip1p contributed more significantly to the restoration of mtDNA synthesis than did import of newly synthesized Mip1p from the cytosol. These observations suggest an important link between function of mitochondrial chaperones and the propagation of mitochondrial genomes under ever-changing environmental conditions.

  15. Bio-forensics

    Energy Technology Data Exchange (ETDEWEB)

    Trewhella, J. (Jill)


    Bioforensics presents significant technical challenges. Determining if an outbreak is natural or not, and then providing evidence to trace an outbreak to its origin is very complex. Los Alamos scientists pioneered research and development that has generated leading edge strain identification methods based on sequence data. Molecular characterization of environmental background samples enable development of highly specific pathogen signatures. Economic impacts of not knowing the relationships at the molecular level Many different kinds of data are needed for DNA-based bio-forensics.

  16. Bioética


    Nunes, Cássia Regina Rodrigues [UNESP; Nunes,Amauri Porto


    A bioética é um novo campo de conhecimento e uma corrente de pensamento que vem se desenvolvendo amplamente no Brasil, principalmente a partir da década de 90. Nesse artigo buscamos contextualizar o seu surgimento, justificando a necessidade da criação desse neologismo, uma vez que a ética estava de certo modo esquecida, em relação à técnica, restringindo-se apenas à ética profissional. Hoje nos deparamos com uma ampla gama de produções nessa área que ajudam a nortear nossa ação enquanto cida...

  17. Tumor-specific Hsp70 plasma membrane localization is enabled by the glycosphingolipid Gb3.

    Directory of Open Access Journals (Sweden)

    Mathias Gehrmann

    Full Text Available BACKGROUND: Human tumors differ from normal tissues in their capacity to present Hsp70, the major stress-inducible member of the HSP70 family, on their plasma membrane. Membrane Hsp70 has been found to serve as a prognostic indicator of overall patient survival in leukemia, lower rectal and non small cell lung carcinomas. Why tumors, but not normal cells, present Hsp70 on their cell surface and the impact of membrane Hsp70 on cancer progression remains to be elucidated. METHODOLOGY/PRINCIPAL FINDINGS: Although Hsp70 has been reported to be associated with cholesterol rich microdomains (CRMs, the partner in the plasma membrane with which Hsp70 interacts has yet to be identified. Herein, global lipid profiling demonstrates that Hsp70 membrane-positive tumors differ from their membrane-negative counterparts by containing significantly higher amounts of globotriaoslyceramide (Gb3, but not of other lipids such as lactosylceramide (LacCer, dodecasaccharideceramide (DoCer, galactosylceramide (GalCer, ceramide (Cer, or the ganglioside GM1. Apart from germinal center B cells, normal tissues are Gb3 membrane-negative. Co-localization of Hsp70 and Gb3 was selectively determined in Gb3 membrane-positive tumor cells, and these cells were also shown to bind soluble Hsp70-FITC protein from outside in a concentration-dependent manner. Given that the latter interaction can be blocked by a Gb3-specific antibody, and that the depletion of globotriaosides from tumors reduces the amount of membrane-bound Hsp70, we propose that Gb3 is a binding partner for Hsp70. The in vitro finding that Hsp70 predominantly binds to artificial liposomes containing Gb3 (PC/SM/Chol/Gb3, 17/45/33/5 confirms that Gb3 is an interaction partner for Hsp70. CONCLUSIONS/SIGNIFICANCE: These data indicate that the presence of Gb3 enables anchorage of Hsp70 in the plasma membrane of tumors and thus they might explain tumor-specific membrane localization of Hsp70.

  18. Chaperonopathies and chaperonotherapy. Hsp60 as therapeutic target in cancer: potential benefits and risks. (United States)

    Cappello, Francesco; Angileri, Francesca; de Macario, Everly Conway; Macario, Alberto J L


    In this minireview we focus on Hsp60 as a target for anticancer therapy. We discuss the new concepts of chaperonopathies and chaperonotherapy and present information on Hsp60 localization in the cell membrane of human tumor cells. We describe novel mechanisms for Hsp60 reaching the extracellular environment that involve membrane-associated stages, as well as data on anti-Hsp60 antibodies found in human sera, both in normal subjects and patients affected by autoimmune diseases. Finally, we discuss possible therapeutic applications of anti-Hsp60 antibodies in cancer treatment, evaluating also side effects on non-tumor cells. In conclusion, the way for investigating Hsp60-targeted anti-tumor therapy is open, at least for those tumors that express Hsp60 on its surface and/or secrete it outside the cell, as is the search for the molecular mechanisms involved in Hsp60 translocation from cytosol to cell membrane: elucidation of this mechanism will greatly facilitate the optimization of chaperonotherapy centered on Hsp60 with anti-tumor efficacy and minimal side effects.

  19. Hsp70 chaperone systems: diversity of cellular functions and mechanism of action. (United States)

    Mayer, M P; Bukau, B


    Hsp70 chaperone systems play an essential role in the life cycle of many proteins not only in an hostile environment but also under normal growth conditions. In the course of evolution the diversification of functions was accompanied by an amplification of components of the Hsp70 system. Here strategies are reviewed how different Hsp70 systems work independently or cooperate with each other in a functional network to perform their housekeeping tasks even under stress conditions. We further discuss how co-chaperones which act as targeting factors regulate the cycle of substrate binding and release upon which the Hsp70 chaperone activity depends.

  20. Functional characterization of the small heat shock protein Hsp12p from Candida albicans.

    Directory of Open Access Journals (Sweden)

    Man-Shun Fu

    Full Text Available Hsp12p is considered to be a small heat shock protein and conserved among fungal species. To investigate the expression of this heat shock protein in the fungal pathogen Candida albicans we developed an anti-CaHsp12p antibody. We show that this protein is induced during stationary phase growth and under stress conditions including heat shock, osmotic, oxidative and heavy metal stress. Furthermore, we find that CaHsp12p expression is influenced by the quorum sensing molecule farnesol, the change of CO(2 concentration and pH. Notably we show that the key transcription factor Efg1p acts as a positive regulator of CaHsp12p in response to heat shock and oxidative stress and demonstrate that CaHsp12p expression is additionally modulated by Hog1p and the cAMP-PKA signaling pathway. To study the function of Hsp12p in C. albicans we generated a null mutant, in which all four CaHSP12 genes have been deleted. Phenotypic analysis of the strain shows that CaHSP12 is not essential for stress resistance, morphogenesis or virulence when tested in a Drosophila model of infection. However, when overexpressed, CaHSP12 significantly enhanced cell-cell adhesion, germ tube formation and susceptibility to azole antifungal agents whilst desensitizing C. albicans to the quorum sensing molecule farnesol.

  1. Cord blood CD4+ T cells respond to self heat shock protein 60 (HSP60.

    Directory of Open Access Journals (Sweden)

    Joost A Aalberse

    Full Text Available BACKGROUND: To prevent harmful autoimmunity most immune responses to self proteins are controlled by central and peripheral tolerance. T cells specific for a limited set of self-proteins such as human heat shock protein 60 (HSP60 may contribute to peripheral tolerance. It is not known whether HSP60-specific T cells are present at birth and thus may play a role in neonatal tolerance. We studied whether self-HSP60 reactive T cells are present in cord blood, and if so, what phenotype these cells have. METHODOLOGY/PRINCIPAL FINDINGS: Cord blood mononuclear cells (CBMC of healthy, full term neonates (n = 21, were cultured with HSP60 and Tetanus Toxoid (TT to study antigen specific proliferation, cytokine secretion and up-regulation of surface markers. The functional capacity of HSP60-induced T cells was determined with in vitro suppression assays. Stimulation of CBMC with HSP60 led to CD4(+ T cell proliferation and the production of various cytokines, most notably IL-10, Interferon-gamma, and IL-6. HSP60-induced T cells expressed FOXP3 and suppressed effector T cell responses in vitro. CONCLUSION: Self-reactive HSP60 specific T cells are already present at birth. Upon stimulation with self-HSP60 these cells proliferate, produce cytokines and express FOXP3. These cells function as suppressor cells in vitro and thus they may be involved in the regulation of neonatal immune responses.

  2. Potentiated Hsp104 variants suppress toxicity of diverse neurodegenerative disease-linked proteins

    Directory of Open Access Journals (Sweden)

    Meredith E. Jackrel


    Full Text Available Protein misfolding is implicated in numerous lethal neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS and Parkinson disease (PD. There are no therapies that reverse these protein-misfolding events. We aim to apply Hsp104, a hexameric AAA+ protein from yeast, to target misfolded conformers for reactivation. Hsp104 solubilizes disordered aggregates and amyloid, but has limited activity against human neurodegenerative disease proteins. Thus, we have previously engineered potentiated Hsp104 variants that suppress aggregation, proteotoxicity and restore proper protein localization of ALS and PD proteins in Saccharomyces cerevisiae, and mitigate neurodegeneration in an animal PD model. Here, we establish that potentiated Hsp104 variants possess broad substrate specificity and, in yeast, suppress toxicity and aggregation induced by wild-type TDP-43, FUS and α-synuclein, as well as missense mutant versions of these proteins that cause neurodegenerative disease. Potentiated Hsp104 variants also rescue toxicity and aggregation of TAF15 but not EWSR1, two RNA-binding proteins with a prion-like domain that are connected with the development of ALS and frontotemporal dementia. Thus, potentiated Hsp104 variants are not entirely non-specific. Indeed, they do not unfold just any natively folded protein. Rather, potentiated Hsp104 variants are finely tuned to unfold proteins bearing short unstructured tracts that are not recognized by wild-type Hsp104. Our studies establish the broad utility of potentiated Hsp104 variants.

  3. Production and preliminary evaluation of Trypanosoma evansi HSP70 for antibody detection in Equids. (United States)

    Kumar, Jaideep; Chaudhury, Ashok; Bera, Bidhan C; Kumar, Ritesh; Kumar, Rajender; Tatu, Utpal; Yadav, Suresh Chandra


    The present immuno-diagnostic method using soluble antigens from whole cell lysate antigen for trypanosomosis have certain inherent problems like lack of standardized and reproducible antigens, as well as ethical issues due to in vivo production, that could be alleviated by in vitro production. In the present study we have identified heat shock protein 70 (HSP70) from T. evansi proteome. The nucleotide sequence of T. evansi HSP70 was 2116 bp, which encodes 690 amino acid residues. The phylogenetic analysis of T. evansi HSP70 showed that T. evansi occurred within Trypanosoma clade and is most closely related to T. brucei brucei and T. brucei gambiense, whereas T. congolense HSP70 laid in separate clade. The two partial HSP70 sequences (HSP-1 from N-terminal region and HSP-2 from C-terminal region) were expressed and evaluated as diagnostic antigens using experimentally infected equine serum samples. Both recombinant proteins detected antibody in immunoblot using serum samples from experimental infected donkeys with T. evansi. Recombinant HSP-2 showed comparable antibody response to Whole cell lysate (WCL) antigen in immunoblot and ELISA. The initial results indicated that HSP70 has potential to detect the T. evansi infection and needs further validation on large set of equine serum samples.

  4. Investigation of HSP60 gene expression in mRNA level in heart at dilated cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Riabenko D. V.


    Full Text Available The expression of HSP60 in the mRNA level in human hearts at the end-stage of dilated cardiomyopathy (DCM as well as in the hearts of mice with disease model similar to human DCM was investigated. We observed a significant increase in the Hsp60 mRNA level at the beginning of the disease and decrease to a normal level at the end stage. As the Hsp60 level was increased during the disease up to the end stage we can presume some changes in the regulation of Hsp60 synthesis or its degradation at DCM progression

  5. Cloning and Sequence Analysis of Hsp90 and Hsp70 Gene from Carposina sasakii Matsumura under Heat Stress%高温胁迫下桃小食心虫热激蛋白Hsp90和Hsp70基因的克隆及序列分析

    Institute of Scientific and Technical Information of China (English)

    高萍; 贾向风; 王洪平


    桃小食心虫(Carposina sasakii Matsumura)是果树上重要的食心虫类害虫,热激蛋白Hsp90和Hsp70在昆虫抵御温度胁迫反应中具有重要作用。采用RT-PCR方法克隆获得了高温胁迫下桃小食心虫热激蛋白Hsp90和Hsp70基因cDNA部分序列,并对其进行分析,为深入揭示桃小食心虫对环境适应的分子机理提供理论依据。已获得的桃小食心虫热激蛋白Hsp90基因(GenBank登录号:KJ139642)序列长420bp,编码139个氨基酸残基,推导的氨基酸序列中含有热激蛋白90家族的一段序列为YSNKEIFLRE的特征序列,并且cDNA序列在N端具有Hsp90基因保守的ATPase结构,该序列与天蚕(Antheraea yamamai)和柞蚕(Antheraea pernyi)等昆虫的氨基酸序列一致性高达99%。已获得的Hsp70-1基因(GenBank登录号:KJ139643)和Hsp70-2基因(GenBank登录号:KJ139644)序列长均为305bp,编码101个氨基酸残基。 Hsp70-1基因推导的氨基酸序列与家蚕(Bombyx mori)的氨基酸序列一致性为94%,Hsp70-2基因推导的氨基酸序列与小菜蛾(Plutella xylostella)和烟草夜蛾(Manduca sexta)等昆虫的氨基酸序列一致性为96%。%Peach fruit moth,Carposina sasakii Matsumura is one of the main pests of the fruit trees. The heat shock protein 90 (Hsp90)and the heat shock protein70(Hsp70)widely exist in insects and play important role under temperature stress. In order to provide theoretical basis for exploring molecular mechanism of insect adaption to environment, partial-length cDNA encoding Hsp90 and Hsp70 from Carposina Sasakii were cloned by RT-PCR and the sequence was analyzed. Sequence analysis indicated that the partial cDNA of Hsp90 (GenBank accession number: KJ139642) was 420bp, encoding 139 amino acid residues. The deduced amino acid sequence contains an important Hsp90 family characteristic sequence(YSNKEIFLRE) and conservative ATPase structure in N-terminal domain, which shares 99% amino acid sequence identies with the Hsp90 genes from

  6. 两种粉虱Hsp60基因的克隆与表达%Cloning and Expression of Hsp60 Gene in Two Whitefly

    Institute of Scientific and Technical Information of China (English)

    郭莉莉; 肖林云; 余昊; 王运兵


    以近缘昆虫Hsp60基因保守区域设计兼并引物,PCR扩增烟粉虱(Bemisiatabaci)地中海(MED)隐种与温室白粉虱(Trialeurodes vaporariorum)Hsp60基因cDNA,并检测了Hsp60基因受温度影响的表达情况.结果表明,烟粉虱MED隐种Hsp60基因cDNA的开放性阅读框长1 821bp,编码607个氨基酸;温室白粉虱Hsp60基因cDNA的开放性阅读框长1 788 bp,编码596个氨基酸,Hsp60基因在昆虫纲低级阶元水平和高级阶元水平系统进化上能得到一个较理想结果.温室白粉虱Hsp60基因表达量受温度影响显著(P<0.05),而烟粉虱MED隐种则不显著(P>0.05).

  7. Bio-tribology. (United States)

    Dowson, Duncan


    It is now forty six years since the separate topics of friction, lubrication, wear and bearing design were integrated under the title 'Tribology' [Department of Education and Science, Lubrication (Tribology) Education and Research. A Report on the Present Position and Industry's Needs, HMSO, London, 1966]. Significant developments have been reported in many established and new aspects of tribology during this period. The subject has contributed to improved performance of much familiar equipment, such as reciprocating engines, where there have been vast improvements in engine reliability and efficiency. Nano-tribology has been central to remarkable advances in information processing and digital equipment. Shortly after widespread introduction of the term tribology, integration with biology and medicine prompted rapid and extensive interest in the fascinating sub-field now known as Bio-tribology [D. Dowson and V. Wright, Bio-tribology, in The Rheology of Lubricants, ed. T. C. Davenport, Applied Science Publishers, Barking, 1973, pp. 81-88]. An outline will be given of some of the developments in the latter field.

  8. Specific recognition of the collagen triple helix by chaperone HSP47. II. The HSP47-binding structural motif in collagens and related proteins. (United States)

    Koide, Takaki; Nishikawa, Yoshimi; Asada, Shinichi; Yamazaki, Chisato M; Takahara, Yoshifumi; Homma, Daisuke L; Otaka, Akira; Ohtani, Katsuki; Wakamiya, Nobutaka; Nagata, Kazuhiro; Kitagawa, Kouki


    The endoplasmic reticulum-resident chaperone heat-shock protein 47 (HSP47) plays an essential role in procollagen biosynthesis. The function of HSP47 relies on its specific interaction with correctly folded triple-helical regions comprised of Gly-Xaa-Yaa repeats, and Arg residues at Yaa positions have been shown to be important for this interaction. The amino acid at the Yaa position (Yaa(-3)) in the N-terminal-adjoining triplet containing the critical Arg (defined as Arg(0)) was also suggested to be directly recognized by HSP47 (Koide, T., Asada, S., Takahara, Y., Nishikawa, Y., Nagata, K., and Kitagawa, K. (2006) J. Biol. Chem. 281, 3432-3438). Based on this finding, we examined the relationship between the structure of Yaa(-3) and HSP47 binding using synthetic collagenous peptides. The results obtained indicated that the structure of Yaa(-3) determined the binding affinity for HSP47. Maximal binding was observed when Yaa(-3) was Thr. Moreover, the required relative spatial arrangement of these key residues in the triple helix was analyzed by taking advantage of heterotrimeric collagen-model peptides, each of which contains one Thr(-3) and one Arg(0). The results revealed that HSP47 recognizes the Yaa(-3) and Arg(0) residues only when they are on the same peptide strand. Taken together, the data obtained led us to define the HSP47-binding structural epitope in the collagen triple helix and also define the HSP47-binding motif in the primary structure. A motif search against human protein database predicted candidate clients for this molecular chaperone. The search result indicated that not all collagen family proteins require the chaperoning by HSP47.

  9. What is BioOne? (United States)

    Fitzpatrick, Roberta Bronson


    BioOne is a Web-based aggregation of full-text, high-impact bioscience research journals. Most of its titles are published by small societies or non-commercial publishers and have not been previously available in electronic format. This column describes the BioOne database and gives some basic information about the best ways to search its content.

  10. Bio-threat microparticle simulants (United States)

    Farquar, George Roy; Leif, Roald N


    A bio-threat simulant that includes a carrier and DNA encapsulated in the carrier. Also a method of making a simulant including the steps of providing a carrier and encapsulating DNA in the carrier to produce the bio-threat simulant.

  11. Bio-threat microparticle simulants (United States)

    Farquar, George Roy; Leif, Roald


    A bio-threat simulant that includes a carrier and DNA encapsulated in the carrier. Also a method of making a simulant including the steps of providing a carrier and encapsulating DNA in the carrier to produce the bio-threat simulant.

  12. COPD and levels of Hsp70 (HSPA1A) and Hsp27 (HSPB1) in plasma and lymphocytes among coal workers: a case-control study. (United States)

    Cui, Xiuqing; Xing, Jingcai; Liu, Yuewei; Zhou, Yun; Luo, Xin; Zhang, Zhihong; Han, Wenhui; Wu, Tangchun; Chen, Weihong


    This case-control study aimed to investigate whether the levels of Hsp70 (HSPA1A) and Hsp27 (HSPB1) in plasma and lymphocytes were associated with the risk of chronic obstructive pulmonary disease (COPD) among coal workers. A total of 76 COPD cases and 48 age-matched healthy controls from a group of coal workers were included. The case group consisted of 35 COPD patients whose condition was complicated with coal workers' pneumoconiosis (CWP) and 41 COPD patients without CWP. Heat shock proteins (Hsps) in plasma and lymphocytes were detected by ELISA and flow cytometry, respectively. Multiple logistic regression models were applied to estimate the association between Hsp levels and COPD risk. Our results showed that plasma Hsp70 and lymphocyte Hsp27 levels were significantly higher and plasma Hsp27 levels were significantly lower in COPD cases than in controls (p < 0.01). No significant differences in lymphocyte Hsp70 levels were found between COPD cases and the matched subjects. Higher plasma Hsp70 levels (odds ratio (OR) = 13.8, 95 % confidence interval (CI) = 5.7-33.5) and lower plasma Hsp27 levels (OR = 4.6, 95 % CI = 2.0-10.5) were significantly associated with an increased risk of COPD after adjusting for confounders. Higher lymphocyte Hsp27 levels were only associated with an increased risk of COPD with CWP (OR = 6.6, 95 % CI = 2.0-22.1) but not with an increased risk of COPD without CWP (OR = 3.0, 95 % CI = 0.9-8.9). Additionally, there were strong joint effects of different Hsps on COPD risk. These results showed that higher levels of plasma Hsp70 and lower levels of plasma Hsp27 might be associated with an increased risk of COPD among coal workers. They may have the potential to serve as monitoring markers for COPD in coal workers.

  13. Blind cavefish and heat shock protein chaperones: a novel role for hsp90alpha in lens apoptosis. (United States)

    Hooven, Thomas A; Yamamoto, Yoshiyuki; Jeffery, William R


    Lens apoptosis plays a central role in cavefish eye degeneration. Heat shock proteins (hsps) can regulate apoptosis; therefore, we examined the relationship between constitutive hsp70 and hsp90 expression and lens apoptosis. The model system is Astyanax mexicanus, a teleost species consisting of an eyed surface-dwelling (surface fish) form and numerous blind cave-dwelling (cavefish) forms. Optic primordia are formed in the cavefish embryo but they subsequently undergo lens apoptosis, arrest in development and degenerate. Astyanax hsp90 and hsp70 DNAs were isolated to use as probes to compare gene expression during surface fish and cavefish development. Hsp90beta, which encodes one of two hsp90 isoforms, was not expressed in the surface fish or cavefish lens, whereas hsp70 was expressed in the lens of both forms, suggesting that neither is directly involved in lens apoptosis. In contrast, hsp90alpha, the other hsp90 isoform, was expressed in the cavefish but not the surface fish lens. Hsp90alpha expression peaked shortly before the beginning of lens apoptosis in three convergent cavefish populations, suggesting a close relationship with lens apoptosis. The absence of hsp90beta in the lens allowed us to use geldanamycin and radicicol, specific inhibitors of hsp90 chaperone function, to determine whether lens cell death requires hsp90alpha expression. Both inhibitors blocked TUNEL labeling in the cavefish lens, suggesting that hsp90alpha is required for apoptosis. In contrast to their effects on the lens, these inhibitors induced TUNEL labeling in the surface epidermis, presumably due to effects on hsp90beta function, implying that the two-hsp90 isoforms may have contrasting roles in cell survival. We conclude that hsp90alpha plays a novel role in lens apoptosis and cavefish eye degeneration.

  14. Exposure levels to radiation in a nuclear medicine laboratory: measurements with thermoluminescence dosemeters; Niveles de exposicion a la radiacion en un laboratorio de medicina nuclear: mediciones con dosimetros de termoluminiscencia

    Energy Technology Data Exchange (ETDEWEB)

    Ruiz J, A. [Departamento de Medicina Nuclear del Instituto nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico D.F. (Mexico)


    Since 1996 in the Nuclear medicine Department and another routine and research departments of the National Institute of Medical and Nutrition Sciences (INCMNSZ) are working at least with 12 radioactive sources opened and sealed. However, it was unknown if with the Tl personal dosemeters with lithium fluoride crystals (LiF), could be possible to receive information about the equivalent dose for each radionuclide or if there was some radionuclide that by being low energy emissor or beta energy emissor, it did not represent an ionization power sufficient to excite the crystals of the thermoluminescent dosemeters (DTL). In this work the obtained results with control dosemeters of diverse source opened or sealed are shown, which were put at 1 cm of the LiF crystals during a time exposure 18 hours. Therefore the objective of this work is to verify which sources excite or not to DTL crystals and taking the pertinent safety measures for each radionuclide. The conclusion is that the majority of dosemeters were excited by beta or gamma radiation of the radionuclides and that LiF crystals are able to receive equivalent doses until 1200 mSv, being this a guarantee for the staff, that in an any moment could be exposed to high dose in his working day. Also it was corroborated that the radionuclides more energizer are: iodine-131, iodine-125 and sodium-22 while the phosphorus-32 must be managed with careful, since the exposure to hands can result significant, just like the technetium-99m ({sup 99m} Tc). (Author)

  15. New ICRU quantities for the environmental and individual monitoring. Standardization of individual dosemeters by using external beams of photon radiation; Nuevas magnitudes ICRU para la vigilancia radiologica ambiental e individual. Calibracion de dosimetros personales usando haces externos de fotones

    Energy Technology Data Exchange (ETDEWEB)

    Brosed, A.; Delgado, A.; Granados, C. E.


    The quantities introduced by ICRU for the radiological monitoring are commented, specially those implied in individual protection against external photons. A procedure is proposed in order to standardize the individual dosemeters by using the kerma in air references of CIEMAT-JEN. The reference radiation beams are described in connection with ISO standards. Provisional values are selected for the appropriate conversion and correction factors. (Author) 23 refs.

  16. Relative thermoluminescent efficiencies proton/gamma and helium/gamma of high temperature peaks in TLD-100 dosemeters; Eficiencias termoluminiscentes relativas proton/gamma y helio/gamma de picos de alta temperatura en dosimetros TLD-100

    Energy Technology Data Exchange (ETDEWEB)

    Flores M, E.; Avila, O.; Rodriguez V, M. [ININ, 52750 La Marquesa, Estado de Mexico (Mexico); Massillon, J.L.G.; Buenfil A, E.; Ruiz T, C.; Brandan, M.E. [IFUNAM, 04500 Mexico D.F. (Mexico); Gamboa De Buen, I. [ICN-UNAM, 04500 Mexico D.F. (Mexico)


    This work presents measures of relative thermoluminescent efficiency of those high temperature peaks of TLD-100 dosemeters exposed to protons of 1.5 MeV and to helium nuclei of 3 and 7.5 MeV. A rigorous reading and of deconvolution protocol was used for the calculation of the TL efficiencies. Additionally an Excel program that facilitated the deconvolution adjustment process of the glow curves was elaborated. (Author)

  17. Diversity of cytosolic HSP70 Heat Shock Protein from decapods and their phylogenetic placement within Arthropoda. (United States)

    Baringou, Stephane; Rouault, Jacques-Deric; Koken, Marcel; Hardivillier, Yann; Hurtado, Luis; Leignel, Vincent


    The 70kDa heat shock proteins (HSP70) are considered the most conserved members of the HSP family. These proteins are primordial to the cell, because of their implications in many cellular pathways (e. g., development, immunity) and also because they minimize the effects of multiple stresses (e. g., temperature, pollutants, salinity, radiations). In the cytosol, two ubiquitous HSP70s with either a constitutive (HSC70) or an inducible (HSP70) expression pattern are found in all metazoan species, encoded by 5 or 6 genes (Drosophila melanogaster or yeast and human respectively). The cytosolic HSP70 protein family is considered a major actor in environmental adaptation, and widely used in ecology as an important biomarker of environmental stress. Nevertheless, the diversity of cytosolic HSP70 remains unclear amongst the Athropoda phylum, especially within decapods. Using 122 new and 311 available sequences, we carried out analyses of the overall cytosolic HSP70 diversity in arthropods (with a focus on decapods) and inferred molecular phylogenies. Overall structural and phylogenetic analyses showed a surprisingly high diversity in cytosolic HSP70 and revealed the existence of several unrecognised groups. All crustacean HSP70 sequences present signature motifs and molecular weights characteristic of non-organellar HSP70, with multiple specific substitutions in the protein sequence. The cytosolic HSP70 family in arthropods appears to be constituted of at least three distinct groups (annotated as A, B and C), which comprise several subdivisions, including both constitutive and inducible forms. Group A is constituted by several classes of Arthropods, while group B and C seem to be specific to Malacostraca and Hexapoda/Chelicerata, respectively. The HSP70 organization appeared much more complex than previously suggested, and far beyond a simple differentiation according to their expression pattern (HSC70 versus HSP70). This study proposes a new classification of cytosolic

  18. Decreased Hsp90 expression in infiltrative lobular carcinoma: an immunohistochemical study

    Directory of Open Access Journals (Sweden)

    Zagouri Flora


    Full Text Available Abstract Background Elevated Hsp90 expression has been documented in breast ductal carcinomas, whereas decreased Hsp90 expression has been reported in precursor lobular lesions. This study aims to assess Hsp90 expression in infiltrative lobular carcinomas of the breast. Methods Tissue specimens were taken from 32 patients with infiltrative lobular carcinoma. Immunohistochemical assessment of Hsp90 was performed both in the lesion and the adjacent normal breast ducts and lobules; the latter serving as control. Concerning Hsp90 assessment: i the percentage of positive cells and ii the intensity were separately analyzed. Subsequently, the Allred score was adopted and calculated. The intensity was treated as an ordinal variable-score (0: negative, low: 1, moderate: 2, high: 3. Statistical analysis followed. Results All infiltrative lobular carcinoma foci mainly presented with a positive cytoplasmic immunoreaction for Hsp90. Compared to the adjacent normal ducts and lobules, infiltrative lobular carcinoma exhibited a statistically significant decrease in Hsp90 expression, both in terms of Hsp90 positive cells (% and Allred score (74.2 ± 11.2 vs. 59.1 ± 14.2 p = 0.0001; 7.00 ± 0.95 vs. 6.22 ± 1.01, p = 0.007, Wilcoxon matched-pairs signed-ranks test. Concerning the intensity of Hsp90 immunostaining only a marginal decrease was noted (2.16 ± 0.68 vs. 1.84 ± 0.63, p = 0.087, Wilcoxon matched-pairs signed-ranks test. Conclusion ILC lesions seem to exhibit decreased Hsp90 expression, a finding contrary to what might have been expected, given that high Hsp90 expression is a trait of invasive ductal carcinomas.

  19. Cytosolic HSP90 Cochaperones HOP and FKBP Interact with Freshly Synthesized Chloroplast Preproteins of Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Christine Fellerer; Regina Schweiger; Katharina Sch(o)ngruber; Jürgen Soll; Serena Schwenkert


    Most chloroplast and mitochondrial proteins are synthesized in the cytosol of the plant cell and have to be imported into the organelles post-translationally.Molecular chaperones play an important role in preventing protein aggregation of freshly translated preproteins and assist in maintaining the preproteins in an import competent state.Preproteins can associate with HSPT0,HSP90,and 14-3-3 proteins in the cytosol.In this study,we analyzed a large set of wheat germ-translated chloroplast preproteins with respect to their chaperone binding.Our results demonstrate that the formation of distinct 14-3-3 or HSP90 containing preprotein complexes is a common feature in post-translational protein transport in addition to preproteins that seem to interact solely with HSP70.We were able to identify a diverse and extensive class of preproteins as HSP90 substrates,thus providing a tool for the investigation of HSP90 client protein association.The analyses of chimeric HSP90 and 14-3-3 binding preproteins with exchanged transit peptides indicate an involvement of both the transit peptide and the mature part of the proteins,in HSP90 binding.We identified two partner components of the HSP90 cycle,which were present in the preprotein containing high-molecular-weight complexes,the HSP70/HSP90 organizing protein HOP,as well as the immunophilin FKBP73.The results establish chloroplast preproteins as a general class of HSP90 client proteins in plants using HOP and FKBP as novel cochaperones.

  20. The complex evolutionary dynamics of Hsp70s: a genomic and functional perspective. (United States)

    Kominek, Jacek; Marszalek, Jaroslaw; Neuvéglise, Cécile; Craig, Elizabeth A; Williams, Barry L


    Hsp70 molecular chaperones are ubiquitous. By preventing aggregation, promoting folding, and regulating degradation, Hsp70s are major factors in the ability of cells to maintain proteostasis. Despite a wealth of functional information, little is understood about the evolutionary dynamics of Hsp70s. We undertook an analysis of Hsp70s in the fungal clade Ascomycota. Using the well-characterized 14 Hsp70s of Saccharomyces cerevisiae, we identified 491 orthologs from 53 genomes. Saccharomyces cerevisiae Hsp70s fall into seven subfamilies: four canonical-type Hsp70 chaperones (SSA, SSB, KAR, and SSC) and three atypical Hsp70s (SSE, SSZ, and LHS) that play regulatory roles, modulating the activity of canonical Hsp70 partners. Each of the 53 surveyed genomes harbored at least one member of each subfamily, and thus establishing these seven Hsp70s as units of function and evolution. Genomes of some species contained only one member of each subfamily that is only seven Hsp70s. Overall, members of each subfamily formed a monophyletic group, suggesting that each diversified from their corresponding ancestral gene present in the common ancestor of all surveyed species. However, the pattern of evolution varied across subfamilies. At one extreme, members of the SSB subfamily evolved under concerted evolution. At the other extreme, SSA and SSC subfamilies exhibited a high degree of copy number dynamics, consistent with a birth-death mode of evolution. KAR, SSE, SSZ, and LHS subfamilies evolved in a simple divergent mode with little copy number dynamics. Together, our data revealed that the evolutionary history of this highly conserved and ubiquitous protein family was surprising complex and dynamic.

  1. Hsp90 as a "Chaperone" of the Epigenome: Insights and Opportunities for Cancer Therapy. (United States)

    Isaacs, Jennifer S


    The cellular functions of Hsp90 have historically been attributed to its ability to chaperone client proteins involved in signal transduction. Although numerous stimuli and the signaling cascades they activate contribute to cancer progression, many of these pathways ultimately require transcriptional effectors to elicit tumor-promoting effects. Despite this obvious connection, the majority of studies evaluating Hsp90 function in malignancy have focused upon its regulation of cytosolic client proteins, and particularly members of receptor and/or kinase families. However, in recent years, Hsp90 has emerged as a pivotal orchestrator of nuclear events. Discovery of an expanding repertoire of Hsp90 clients has illuminated a vital role for Hsp90 in overseeing nuclear events and influencing gene transcription. Hence, this chapter will cast a spotlight upon several regulatory themes involving Hsp90-dependent nuclear functions. Highlighted topics include a summary of chaperone-dependent regulation of key transcription factors (TFs) and epigenetic effectors in malignancy, as well as a discussion of how the complex interplay among a subset of these TFs and epigenetic regulators may generate feed-forward loops that further support cancer progression. This chapter will also highlight less recognized indirect mechanisms whereby Hsp90-supported signaling may impinge upon epigenetic regulation. Finally, the relevance of these nuclear events is discussed within the framework of Hsp90's capacity to enable phenotypic variation and drug resistance. These newly acquired insights expanding our understanding of Hsp90 function support the collective notion that nuclear clients are major beneficiaries of Hsp90 action, and their impairment is likely responsible for many of the anticancer effects elicited by Hsp90-targeted approaches.

  2. Catapult mechanism renders the chaperone action of Hsp70 unidirectional. (United States)

    Gisler, S M; Pierpaoli, E V; Christen, P


    Molecular chaperones of the Hsp70 type promote the folding and membrane translocation of proteins. The interaction of Hsp70s with polypeptides is linked to ATP binding and hydrolysis. We formed complexes of seven different fluorescence-labeled peptides with DnaK, the Hsp70 homolog of Escherichia coli, and determined the rate of peptide release under two different sets of conditions. (1) Upon addition of ATP to nucleotide-free peptide.DnaK complexes, all tested peptides were released with similar rate constants (2.2 s-1 to 6.7 s-1). (2) In the binding equilibrium of peptide and ATP-liganded DnaK, the dissociation followed one or two-step reactions, depending on the amino acid sequence of the peptide. For the monophasic reactions, the dissociation rate constants diverged by four orders of magnitude from 0.0004 s-1 to 5.7 s-1; for the biphasic reactions, the rate constants of the second, slower isomerization step were in the range from 0.3 s-1 to 0.0005 s-1. The release of the different peptides in case (1) is 1.4 to 14,000 times faster than in case (2). Apparently, binding of ATP induces a transient state of the chaperone which ejects target peptides before the final state of ATP-liganded DnaK is reached. This "catapult" mechanism provides the chaperone cycle with a mode of peptide release that does not correspond with the reverse of peptide binding. By allowing the conformation of the outgoing polypeptide to differ from that of the incoming polypeptide, a futile cycle with respect to conformational work exerted on the target protein is obviated.

  3. Cardiac myocyte exosomes: stability, HSP60, and proteomics. (United States)

    Malik, Z A; Kott, K S; Poe, A J; Kuo, T; Chen, L; Ferrara, K W; Knowlton, A A


    Exosomes, which are 50- to 100-nm-diameter lipid vesicles, have been implicated in intercellular communication, including transmitting malignancy, and as a way for viral particles to evade detection while spreading to new cells. Previously, we demonstrated that adult cardiac myocytes release heat shock protein (HSP)60 in exosomes. Extracellular HSP60, when not in exosomes, causes cardiac myocyte apoptosis via the activation of Toll-like receptor 4. Thus, release of HSP60 from exosomes would be damaging to the surrounding cardiac myocytes. We hypothesized that 1) pathological changes in the environment, such as fever, change in pH, or ethanol consumption, would increase exosome permeability; 2) different exosome inducers would result in different exosomal protein content; 3) ethanol at "physiological" concentrations would cause exosome release; and 4) ROS production is an underlying mechanism of increased exosome production. We found the following: first, exosomes retained their protein cargo under different physiological/pathological conditions, based on Western blot analyses. Second, mass spectrometry demonstrated that the protein content of cardiac exosomes differed significantly from other types of exosomes in the literature and contained cytosolic, sarcomeric, and mitochondrial proteins. Third, ethanol did not affect exosome stability but greatly increased the production of exosomes by cardiac myocytes. Fourth, ethanol- and hypoxia/reoxygenation-derived exosomes had different protein content. Finally, ROS inhibition reduced exosome production but did not completely inhibit it. In conclusion, exosomal protein content is influenced by the cell source and stimulus for exosome formation. ROS stimulate exosome production. The functions of exosomes remain to be fully elucidated.

  4. Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer. (United States)

    Rappa, Francesca; Pitruzzella, Alessandro; Marino Gammazza, Antonella; Barone, Rosario; Mocciaro, Emanuele; Tomasello, Giovanni; Carini, Francesco; Farina, Felicia; Zummo, Giovanni; Conway de Macario, Everly; Macario, Alberto Jl; Cappello, Francesco


    Large bowel carcinogenesis involves accumulation of genetic alterations leading to transformation of normal mucosa into dysplasia and, lastly, adenocarcinoma. It is pertinent to elucidate the molecular changes occurring in the pre-neoplastic lesions to facilitate early diagnosis and treatment. Heat shock proteins (Hsps), many of which are molecular chaperones, are implicated in carcinogenesis, and their variations with tumor progression encourage their study as biomarkers. There are many reports on Hsps and cancer but none to our knowledge on their systematic quantification in pre-neoplastic lesions of the large bowel. We performed immunohistochemical determinations of Hsp10, Hsp60, Hsp70, and Hsp90 in biopsies of large bowel tubular adenomas with moderate grade of dysplasia and compared to normal mucosa and adenocarcinoma with a moderate grade of differentiation (G2). A significant elevation of Hsp10 and Hsp60 only, i.e., in the absence of elevation of Hsp70 or Hsp90, in both epithelium and lamina propria was found in tubular adenoma by comparison with normal mucosa. In contrast, adenocarcinoma was characterized by the highest levels of Hsp10 and Hsp60 in epithelium and lamina propria, accompanied by the highest levels of Hsp70 only in epithelium and of Hsp90 only in lamina propria, by comparison with normal and tubular adenoma counterparts. Hsp10 and Hsp60 are promising biomarkers for early diagnosis of tubular adenoma and for its differentiation from more advanced malignant lesions. Hsp10 and Hsp60 may be implicated in carcinogenesis from its very early steps and, thus, are potentially convenient targets for therapy.

  5. Targeting Plasmodium falciparum Hsp90: Towards Reversing Antimalarial Resistance

    Directory of Open Access Journals (Sweden)

    Dea Shahinas


    Full Text Available Malaria continues to exact a great human toll in tropical settings. Antimalarial resistance is rife and the parasite inexorably develops mechanisms to outwit our best drugs, including the now first-line choice, artesunate. Novel strategies to circumvent resistance are needed. Here we detail drug development focusing on heat shock protein 90 and its central role as a chaperone. A growing body of evidence supports the role for Hsp90 inhibitors as adjunctive drugs able to restore susceptibility to traditionally efficacious compounds like chloroquine.

  6. Bio-ethanol

    DEFF Research Database (Denmark)

    Wenzel, Henrik


    , however, shown that by following this path, we will lose more than we gain on both CO2 emission and fossil fuel dependency. Being renewable, CO2 neutral and storable, biomass is a priority resource for fossil fuel substitution in general. Investigations of the magnitude of biomass that is or can be made...... available for energy purposes - be it from waste, agricultural residues or energy crops - show, however, that biomass is very limited compared to the potential use of it. Even in the most optimistic near term scenarios (30 years ahead), the total physically available biomass can at maximum substitute around......, but they do not improve the energy balance enough for bio-ethanol to compete with alternative uses of the biomass. When using biomass to substitute fossil fuels in heat & power production, a close to 100% substitution efficiency is achieved. The best alternative for CO2 reduction and oil saving is, therefore...

  7. Quercetin suppresses heat shock-induced nuclear translocation of Hsp72

    Directory of Open Access Journals (Sweden)

    Antoni Gawron


    Full Text Available The effect of quercetin and heat shock on the Hsp72 level and distribution in HeLa cells was studied by Western blotting, indirect immunofluorescence and immunogold electron microscopy. In control cells and after quercetin treatment, Hsp72 was located both in the cytoplasm and in the nucleus in comparable amounts. After hyperthermia, the level of nuclear Hsp72 raised dramatically. Expression of Hsp72 in cytoplasm was also higher but not to such extent as that observed in the nucleus. Preincubation of heated cells with quercetin inhibited strong Hsp72 expression observed after hyperthermia and changed the intracellular Hsp72 distribution. The cytoplasmic level of protein exceeded the nuclear one, especially around the nucleus, where the coat of Hsp72 was noticed. Observations indicating that quercetin was present around and in the nuclear envelope suggested an involvement of this drug in the inhibition of nuclear translocation. Our results indicate that pro-apoptotic activity of quercetin may be correlated not only with the inhibition of Hsp72 expression but also with suppression of its migration to the nucleus.

  8. A primate specific extra domain in the molecular chaperone Hsp90.

    Directory of Open Access Journals (Sweden)

    Vishwadeepak Tripathi

    Full Text Available Hsp90 (heat shock protein 90 is an essential molecular chaperone that mediates folding and quality control of client proteins. Many of them such as protein kinases, steroid receptors and transcription factors are involved in cellular signaling processes. Hsp90 undergoes an ATP hydrolysis dependent conformational cycle to assist folding of the client protein. The canonical Hsp90 shows a typical composition of three distinct domains and interacts with individual cochaperone partners such as Hop, Cdc37 and Aha1 (activator of Hsp90 ATPase that regulate the reaction cycle of the molecular chaperone. A bioinformatic survey identified an additional domain of 122 amino acids in front of the canonical Hsp90 sequence. This extra domain (E domain is specific to the Catarrhini or drooping nose monkeys, a subdivision of the higher primates that includes man, the great apes and the old world monkeys but is absent from all other species. Our biochemical analysis reveals that Hsp103 associates with cochaperone proteins such as Hop, Cdc37 and Aha1 similar to Hsp90. However, the extra domain reduces the ATP hydrolysis rate to about half when compared to Hsp90 thereby acting as a negative regulator of the molecular chaperonés intrinsic ATPase activity.

  9. Hsp60, a novel target for antitumor therapy: structure-function features and prospective drugs design. (United States)

    Pace, Andrea; Barone, Giampaolo; Lauria, Antonino; Martorana, Annamaria; Piccionello, Antonio Palumbo; Pierro, Paola; Terenzi, Alessio; Almerico, Anna Maria; Buscemi, Silvestre; Campanella, Claudia; Angileri, Francesca; Carini, Francesco; Zummo, Giovanni; de Macario, Everly Conway; Cappello, Francesco; Macario, Alberto J L


    Heat shock protein 60 kDa (Hsp60) is a chaperone classically believed to be involved in assisting the correct folding of other mitochondrial proteins. Hsp60 also plays a role in cytoprotection against cell stressors, displaying for example, antiapoptotic potential. Despite the plethora of studies devoted to the mechanism of Hsp60's function, especially in prokaryotes, fundamental issues still remain unexplored, including the definition of its role in cancer. Key questions still unanswered pertain to the differences in structure-function features that might exist between the well-studied prokaryotic GroEL and the largely unexplored eukaryotic Hsp60 proteins. In this article we discuss these differences in sequence, structure, and roles of Hsp60, focusing on the human ortholog with the view of devising compounds to block its ability to favour tumor-cell growth and survival. Compounds currently known to directly or indirectly affect Hsp60 functions, such as protein folding, HIF-1α accumulation, or Hsp60-induced cell proliferation, are discussed along with strategies that might prove effective for developing Hsp60-targeting drugs for anticancer therapy.

  10. Spermatogenesis arrest caused by conditional deletion of Hsp90α in adult mice

    Directory of Open Access Journals (Sweden)

    Chiaki Kajiwara


    It is controversial whether a functional androgen receptor (AR on germ cells, including spermatogonia, is essential for their development into sperm and, thus, initiation and maintenance of spermatogenesis. It was recently shown that many spermatocytes underwent apoptosis in the testes of Hsp90α KO mice. We had generated Hsp90α KO mice independently and confirmed this phenotype. However, the important question of whether Hsp90α is required to maintain spermatogenesis in adult mice in which testicular maturation is already completed could not be addressed using these conventional KO mice. To answer this question, we generated a tamoxifen-inducible deletion mutant of Hsp90α and found that conditional deletion of Hsp90α in adult mice caused even more severe apoptosis in germ cells beyond the pachytene stage, leading to complete arrest of spermatogenesis and testicular atrophy. Importantly, immunohistochemical analysis revealed that AR expression in WT testis was more evident in spermatogonia than in spermatocytes, whereas its expression was aberrant and ectopic in Hsp90α KO testis, raising the possibility that an AR abnormality in primordial germ cells is involved in spermatogenesis arrest in the Hsp90α KO mice. Our results suggest that the AR, specifically chaperoned by Hsp90α in spermatogonia, is critical for maintenance of established spermatogenesis and for survival of spermatocytes in adult testis, in addition to setting the first wave of spermatogenesis before puberty.

  11. Functional analysis of the Hikeshi-like protein and its interaction with HSP70 in Arabidopsis

    Energy Technology Data Exchange (ETDEWEB)

    Koizumi, Shinya; Ohama, Naohiko; Mizoi, Junya [Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan); Shinozaki, Kazuo [RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama, Kanagawa 230-0045 (Japan); Yamaguchi-Shinozaki, Kazuko, E-mail: [Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657 (Japan)


    Highlights: • HKL, a Hikeshi homologous gene is identified in Arabidopsis. • HKL interacts with two HSP70 isoforms and regulates the subcellular localization of HSC70-1. • The two HSP70 translocate into nucleus in response to heat stress. • Overexpression of HKL confers thermotolerance in transgenic plants. - Abstract: Heat shock proteins (HSPs) refold damaged proteins and are an essential component of the heat shock response. Previously, the 70 kDa heat shock protein (HSP70) has been reported to translocate into the nucleus in a heat-dependent manner in many organisms. In humans, the heat-induced translocation of HSP70 requires the nuclear carrier protein Hikeshi. In the Arabidopsis genome, only one gene encodes a protein with high homology to Hikeshi, and we named this homolog Hikeshi-like (HKL) protein. In this study, we show that two Arabidopsis HSP70 isoforms accumulate in the nucleus in response to heat shock and that HKL interacts with these HSP70s. Our histochemical analysis revealed that HKL is predominantly expressed in meristematic tissues, suggesting the potential importance of HKL during cell division in Arabidopsis. In addition, we show that HKL regulates HSP70 localization, and HKL overexpression conferred thermotolerance to transgenic Arabidopsis plants. Our results suggest that HKL plays a positive role in the thermotolerance of Arabidopsis plants and cooperatively interacts with HSP70.

  12. The FNIP co-chaperones decelerate the Hsp90 chaperone cycle and enhance drug binding (United States)

    Woodford, Mark R.; Dunn, Diana M.; Blanden, Adam R.; Capriotti, Dante; Loiselle, David; Prodromou, Chrisostomos; Panaretou, Barry; Hughes, Philip F.; Smith, Aaron; Ackerman, Wendi; Haystead, Timothy A.; Loh, Stewart N.; Bourboulia, Dimitra; Schmidt, Laura S.; Marston Linehan, W.; Bratslavsky, Gennady; Mollapour, Mehdi


    Heat shock protein-90 (Hsp90) is an essential molecular chaperone in eukaryotes involved in maintaining the stability and activity of numerous signalling proteins, also known as clients. Hsp90 ATPase activity is essential for its chaperone function and it is regulated by co-chaperones. Here we show that the tumour suppressor FLCN is an Hsp90 client protein and its binding partners FNIP1/FNIP2 function as co-chaperones. FNIPs decelerate the chaperone cycle, facilitating FLCN interaction with Hsp90, consequently ensuring FLCN stability. FNIPs compete with the activating co-chaperone Aha1 for binding to Hsp90, thereby providing a reciprocal regulatory mechanism for chaperoning of client proteins. Lastly, downregulation of FNIPs desensitizes cancer cells to Hsp90 inhibitors, whereas FNIPs overexpression in renal tumours compared with adjacent normal tissues correlates with enhanced binding of Hsp90 to its inhibitors. Our findings suggest that FNIPs expression can potentially serve as a predictive indicator of tumour response to Hsp90 inhibitors. PMID:27353360

  13. HSP10 selective preference for myeloid and megakaryocytic precursors in normal human bone marrow

    Directory of Open Access Journals (Sweden)

    F Cappello


    Full Text Available Heat shock proteins (HSPs constitute a heterogeneous family of proteins involved in cell homeostasis. During cell life they are involved in harmful insults, as well as in immune and inflammatory reactions. It is known that they regulate gene expression, and cell proliferation, differentiation and death. HSP60 is a mitochondrial chaperonin, highly preserved during evolution, responsible of protein folding. Its function is strictly dependent on HSP10 in both prokaryotic and eukaryotic elements. We investigated the presence and the expression of HSP60 and HSP10 in a series of 20 normal human bone marrow specimens (NHBM by the means of immunohistochemistry. NHBM showed no expression of HSP60, probably due to its being below the detectable threshold, as already demonstrated in other normal human tissues. By contrast, HSP10 showed a selective positivity for myeloid and megakaryocytic lineages. The positivity was restricted to precursor cells, while mature elements were constantly negative.We postulate that HSP10 plays a role in bone marrow cell differentiation other than being a mitochondrial co-chaperonin. The present data emphasize the role of HSP10 during cellular homeostasis and encourage further investigations in this field.

  14. Expression profile of HSP genes during different seasons in goats (Capra hircus). (United States)

    Dangi, Satyaveer Singh; Gupta, Mahesh; Maurya, Divakar; Yadav, Vijay Prakash; Panda, Rudra Prasanna; Singh, Gyanendra; Mohan, Nitai Haridas; Bhure, Sanjeev Kumar; Das, Bikash Chandra; Bag, Sadhan; Mahapatra, Ramkrishna; Taru Sharma, Guttalu; Sarkar, Mihir


    The present study has demonstrated the expression of HSP60, HSP70, HSP90, and UBQ in peripheral blood mononuclear cells (PBMCs) during different seasons in three different age groups (Groups I, II, and III with age of 0-2, 2-5, and >5 years, respectively) of goats of tropical and temperate regions. Real-time polymerase chain reaction was applied to investigate mRNA expression of examined factors. Specificity of the desired products was documented using analysis of the melting temperature and high-resolution gel electrophoresis to verify that the transcripts are of the exact molecular size predicted. The mRNA expression of HSP60, HSP90, and UBQ was significantly higher (P < 0.05) in all age groups during peak summer season as compared with peak winter season in both tropical and temperate region goats. HSP70 mRNA expression was significantly higher (P < 0.05) during summer season as compared with winter season in tropical region goats. However, in the temperate region, in goats from all the three age groups studied, a non-significant difference of HSP70 expression between summer and winter seasons was noticed. In conclusion, results demonstrate that (1) HSP genes are expressed in caprine PBMCs and (2) higher expression of HSPs during thermal stress suggest possible involvement of them to ameliorate deleterious effect of thermal stress so as to maintain cellular integrity and homeostasis in goats.

  15. Analysis of Hsp90 cochaperone interactions reveals a novel mechanism for TPR protein recognition. (United States)

    Chadli, Ahmed; Bruinsma, Elizabeth S; Stensgard, Bridget; Toft, David


    The chaperone Hsp90 is required for the appropriate regulation of numerous key signaling molecules, including the progesterone receptor (PR). Many important cochaperones bind Hsp90 through their tetratricopeptide repeat (TPR) domains. Two such proteins, GCUNC45 and FKBP52, assist PR chaperoning and are thought to interact sequentially with PR-Hsp90 complexes. TPR proteins bind to the C-terminal MEEVD sequence of Hsp90, but GCUNC45 has been shown also to bind to a novel site near the N-terminus. We now show that FKBP52 is also able to bind to this site, and that these two cochaperones act competitively, through Hsp90, to modulate PR activity. The N-terminal site involves noncontiguous amino acids within or near the ATP binding pocket of Hsp90. TPR interactions at this site are thus strongly regulated by nucleotide binding and Hsp90 conformation. We propose an expanded model for client chaperoning in which the coordinated use of TPR recognition sites at both N- and C-terminal ends of Hsp90 enhances its ability to coordinate interactions with multiple TPR partners.

  16. Why is this effective HSP90 inhibitor not being developed in HER2+ breast cancer? (United States)

    Arteaga, Carlos L


    Inhibition of the HSP90 chaperone leads to degradation of the HER2 receptor. The HSP90 inhibitor tanespimycin in combination with trastuzumab is active in patients with HER2-overexpressing metastatic breast cancer. This combination is one of several HER2-targeted therapies that will significantly improve the outcome of patients with this subtype of breast cancer.

  17. Inhibition of hsp70 by methylene blue affects signaling protein function and ubiquitination and modulates polyglutamine protein degradation. (United States)

    Wang, Adrienne M; Morishima, Yoshihiro; Clapp, Kelly M; Peng, Hwei-Ming; Pratt, William B; Gestwicki, Jason E; Osawa, Yoichi; Lieberman, Andrew P


    The Hsp90/Hsp70-based chaperone machinery regulates the activity and degradation of many signaling proteins. Cycling with Hsp90 stabilizes client proteins, whereas Hsp70 interacts with chaperone-dependent E3 ubiquitin ligases to promote protein degradation. To probe these actions, small molecule inhibitors of Hsp70 would be extremely useful; however, few have been identified. Here we test the effects of methylene blue, a recently described inhibitor of Hsp70 ATPase activity, in three well established systems of increasing complexity. First, we demonstrate that methylene blue inhibits the ability of the purified Hsp90/Hsp70-based chaperone machinery to enable ligand binding by the glucocorticoid receptor and show that this effect is due to specific inhibition of Hsp70. Next, we establish that ubiquitination of neuronal nitric-oxide synthase by the native ubiquitinating system of reticulocyte lysate is dependent upon both Hsp70 and the E3 ubiquitin ligase CHIP and is blocked by methylene blue. Finally, we demonstrate that methylene blue impairs degradation of the polyglutamine expanded androgen receptor, an Hsp90 client mutated in spinal and bulbar muscular atrophy. In contrast, degradation of an amino-terminal fragment of the receptor, which lacks the ligand binding domain and, therefore, is not a client of the Hsp90/Hsp70-based chaperone machinery, is enhanced through homeostatic induction of autophagy that occurs when Hsp70-dependent proteasomal degradation is inhibited by methylene blue. Our data demonstrate the utility of methylene blue in defining Hsp70-dependent functions and reveal divergent effects on polyglutamine protein degradation depending on whether the substrate is an Hsp90 client.

  18. 温室粉虱和烟粉虱3个隐种中热激蛋白基因hsp70和hsp90含量的比较分析%Comparative analysis of the copy numbers of hsp70 and hsp90 in Trialeurodes vaporariorum and three cryptic species of Bemisia tabaci complex (Hemiptera: Aleyrodidae)

    Institute of Scientific and Technical Information of China (English)

    韦姣; 吕志创; 王韧; 万方浩


    [目的]昆虫适应新环境的能力与其对温度的耐受能力密切相关.热激蛋白HSP70和HSP90具有提高生物体温度耐受性的功能.烟粉虱Bemisia tabaci (Gennadius)不同隐种和不同种粉虱对温度的适应能力有差异,这与它们的热激蛋白基因拷贝数的差异可能相关.[方法]利用实时荧光定量PCR方法,检测入侵型烟粉虱MED隐种和MEAM1隐种、本地型烟粉虱AsiaⅡ1隐种以及温室粉虱Trialeurodes vaporariorum (Westwood)基因组DNA中热激蛋白基因hsp70和hsp90的拷贝数.[结果]不同种类的粉虱和烟粉虱不同隐种体内的hsp70和hsp90的含量均有较大差异,其中hsp70和hsp90拷贝数在入侵型烟粉虱MED和MEAM1隐种中含量较其他两种均高,而在土著种AsiaⅡ1隐种中含量最低,在温室粉虱中居中.此外,相同物种雌雄成虫hsp70和hsp90的拷贝数也不同,雌虫hsp70和hsp90拷贝数约为雄虫的2倍.[结论]不同种粉虱及烟粉虱不同隐种的hsp70和hsp90的拷贝数可能与其耐热性差异相关.本研究为解释不同种粉虱、烟粉虱不同隐种及其不同性别的耐热性差异机制提供了进一步的依据.

  19. Increased levels of IgG antibodies against human HSP60 in patients with spondyloarthritis.

    Directory of Open Access Journals (Sweden)

    Astrid Hjelholt

    Full Text Available Spondyloarthritis (SpA comprises a heterogeneous group of inflammatory diseases, with strong association to human leukocyte antigen (HLA-B27. A triggering bacterial infection has been considered as the cause of SpA, and bacterial heat shock protein (HSP seems to be a strong T cell antigen. Since bacterial and human HSP60, also named HSPD1, are highly homologous, cross-reactivity has been suggested in disease initiation. In this study, levels of antibodies against bacterial and human HSP60 were analysed in SpA patients and healthy controls, and the association between such antibodies and disease severity in relation to HLA-B27 was evaluated.Serum samples from 82 patients and 50 controls were analysed by enzyme-linked immunosorbent assay (ELISA for immunoglobulin (IgG1, IgG2, IgG3 and IgG4 antibodies against human HSP60 and HSP60 from Chlamydia trachomatis, Salmonella enteritidis and Campylobacter jejuni. Disease severity was assessed by the clinical scorings Bath Ankylosing Spondylitis Disease Activity Index (BASDAI, Bath Ankylosing Spondylitis Functional Index (BASFI and Bath Ankylosing Spondylitis Metrology Index (BASMI. Levels of IgG1 and IgG3 antibodies against human HSP60, but not antibodies against bacterial HSP60, were elevated in the SpA group compared with the control group. Association between IgG3 antibodies against human HSP60 and BASMI was shown in HLA-B27⁺ patients. Only weak correlation between antibodies against bacterial and human HSP60 was seen, and there was no indication of cross-reaction. These results suggest that antibodies against human HSP60 is associated with SpA, however, the theory that antibodies against human HSP60 is a specific part of the aetiology, through cross-reaction to bacterial HSP60, cannot be supported by results from this study. We suggest that the association between elevated levels of antibodies against human HSP60 and disease may reflect a general activation of the immune system and an increased

  20. Expression of AeaHsp26 and AeaHsp83 in Aedes aegypti (Diptera: Culicidae) Larvae and Pupae in Response to Heat Shock Stress (United States)


    suppressing expression of Hsp23 and Hsp70 in ßies by using RNAi does not alter the decision to enter diapause or the duration of diapause in insects , insects , including mosquitoes, and may have a cu- mulative role in responding to stress induced by ele- vated temperature (Mahroof et al. 2005...response to stress condi- tions other than temperature in human cells andmany other animals and insects including mosquitoes (Mosser et al. 1988

  1. Ectoine from halophilic microorganisms induces the expression of hsp70 and hsp70B' in human keratinocytes modulating the proinflammatory response. (United States)

    Buommino, Elisabetta; Schiraldi, Chiara; Baroni, Adone; Paoletti, Iole; Lamberti, Monica; De Rosa, Mario; Tufano, Maria Antonietta


    The heat shock proteins (Hsps) have an important role in the cytoprotection and repair of cells and tissues. One potential mechanism of protection is the ability of Hsp to inhibit genetic expression of proinflammatory cytokines, the transcription of which is dependent on nuclear factor-kappa B (NF-kappaB) activation. In this study, we evaluated the ability of ectoine, a novel natural biomolecule produced by halophilic microorganisms, to activate the hsp70 and hsp70B'. By reverse transcriptase-polymerase chain reaction and Western blot analysis, we demonstrated increased hsp70B' gene expression in human keratinocytes treated with ectoine and heat stressed. In contrast, in the absence of heat shock, ectoine was unable to induce hsp70B' but had the ability to induce another member of the Hsp family, the hsp70. The latter is not only elevated in response to stress but is also present at basal level in unstressed cells. In addition, ectoine had no effect on proinflammatory cytokines interleukin (IL)-1alpha, IL-6, IL-8, and tumor necrosis factor-alpha and on NF-kappaB and IkappaB-alpha pathway, whereas it downregulated the expression of cited proinflammatory cytokines, in lipopolysaccharides-treated keratinocytes. These results highlighted the ability of ectoine to protect cells from stress conditions and to prevent cell damage by maintaining an elevated level of the Hsp70. Overall, these data might suggest the use of this compatible solute in cosmetic and even pharmaceutical preparations aiming to activate a cytoprotective heat shock response in human cells.

  2. Combining Bio-inspired Sensing with Bio-inspired Locomotion

    DEFF Research Database (Denmark)

    Shaikh, Danish; Hallam, John; Christensen-Dalsgaard, Jakob

    In this paper we present a preliminary Braitenberg vehicle–like approach to combine bio-inspired audition with bio-inspired quadruped locomotion in simulation. Locomotion gaits of the salamander–like robot Salamandra robotica are modified by a lizard’s peripheral auditory system model that modula......In this paper we present a preliminary Braitenberg vehicle–like approach to combine bio-inspired audition with bio-inspired quadruped locomotion in simulation. Locomotion gaits of the salamander–like robot Salamandra robotica are modified by a lizard’s peripheral auditory system model...... that modulates the parameters of the locomotor central pattern generators. We present phonotactic performance results of the simulated lizard-salamander hybrid robot....

  3. Bio-Glasses An Introduction

    CERN Document Server

    Jones, Julian


    This new work is dedicated to glasses and their variants which can be used as biomaterials to repair diseased and damaged tissues. Bio-glasses are superior to other biomaterials in many applications, such as healing bone by signaling stem cells to become bone cells.   Key features:  First book on biomaterials to focus on bio-glassesEdited by a leading authority on bio-glasses trained by one of its inventors, Dr Larry HenchSupported by the International Commission on Glass (ICG)Authored by members of the ICG Biomedical Glass Committee, with the goal of creating a seamless textb

  4. Dexamethasone regulates CFTR expression in Calu-3 cells with the involvement of chaperones HSP70 and HSP90.

    Directory of Open Access Journals (Sweden)

    Luiz Felipe M Prota

    Full Text Available BACKGROUND: Dexamethasone is widely used for pulmonary exacerbation in patients with cystic fibrosis, however, not much is known about the effects of glucocorticoids on the wild-type cystic fibrosis channel transmembrane regulator (CFTR. Our aim was to determine the effects of dexamethasone treatment on wild-type CFTR expression. METHODS AND RESULTS: Dose-response (1 nM to 10 µM and time course (3 to 48 h curves were generated for dexamethasone for mRNA expression in Calu-3 cells using a real-time PCR. Within 24 h, dexamethasone (10 nM showed a 0.3-fold decrease in CFTR mRNA expression, and a 3.2-fold increase in αENaC mRNA expression compared with control groups. Dexamethasone (10 nM induced a 1.97-fold increase in the total protein of wild-type CFTR, confirmed by inhibition by mifepristone. To access surface protein expression, biotinylation followed by Western blotting showed that dexamethasone treatment led to a 2.35-fold increase in the amount of CFTR in the cell surface compared with the untreated control groups. Once protein translation was inhibited with cycloheximide, dexamethasone could not increase the amount of CFTR protein. Protein stability was assessed by inhibition of protein synthesis with cycloheximide (50 µg/ml at different times in cells treated with dexamethasone and in untreated cells. Dexamethasone did not alter the degradation of wild-type CFTR. Assessment of the B band of CFTR within 15 min of metabolic pulse labeling showed a 1.5-fold increase in CFTR protein after treatment with dexamethasone for 24 h. Chaperone 90 (HSP90 binding to CFTR increased 1.55-fold after treatment with dexamethasone for 24 h, whereas chaperone 70 (HSP70 binding decreased 0.30 fold in an immunoprecipitation assay. CONCLUSION: Mature wild-type CFTR protein is regulated by dexamethasone post transcription, involving cotranslational mechanisms with HSP90 and HSP70, which enhances maturation and expression of wild-type CFTR.

  5. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle; Andresen, Lars; Hansen, Karen Aagaard

    , membrane-bound Hsp70 can stimulate antigen presenting cells (APCs) to release proinflammatory cytokines and can provide a target structure for NK cell-mediated lysis. Human cancer cells frequently express Hsp70 on their cell surface, whereas the corresponding normal tissues do not. In addition, several...... clinically applied reagents, such as alkyl-lysophospholipides, chemotherapeutic agents, and anti-inflammatory reagents, have been found to enhance Hsp70 surface expression on cancer cells. We have found that inhibition of histone deacetylase (HDAC) activity leads to surface expression of Hsp70 on various...... hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC-inhibitor mediated Hsp70 surface expression was confined to the apoptotic Annexin V positive cells and blocked by inhibition of apoptosis. Other chemotherapeutic inducers of apoptosis...

  6. Cell-surface expression of Hsp70 on hematopoietic cancer cells after inhibition of HDAC activity

    DEFF Research Database (Denmark)

    Jensen, Helle

    -derived antigenic peptides, a function which is currently explored in immunotherapeutic approaches against cancer. Additionally, membrane-bound Hsp70 can stimulate antigen presenting cells to release proinflammatory cytokines and can provide a target structure for NK cell-mediated lysis. Human cancer cells...... frequently express Hsp70 on their cell surface, whereas the corresponding normal tissues do not. In addition, several clinically applied reagents, such as alkyl-lysophospholipides, chemotherapeutic agents, and anti-inflammatory reagents, have been found to enhance Hsp70 cell surface expression on cancer...... cells. We have found that inhibition of histone deacetylase (HDAC) activity leads to surface expression of Hsp70 on various hematopoietic cancer cells, an occurance that was not observed on naïve or activated peripheral blood cells. HDAC-inhibitor mediated Hsp70 cell surface expression was confined...

  7. Phosphorylation and oligomerization states of native pig brain HSP90 studied by mass spectrometry

    DEFF Research Database (Denmark)

    Garnier, C.; Lafitte, D.; Jorgensen, T.J.;


    HSP90 purified from pig brain. The two protein isoforms were clearly distinguished by ESI-MS, the alpha isoform being approximately six times more abundant than the beta isoform. ESI-MS in combination with lambda phosphatase treatment provided direct evidence of the existence of four phosphorylated...... such as actin-microfilament, tubulin-microtubule and intermediate filaments, and also exhibits conventional chaperone functions. This protein exists in two isoforms alpha-HSP90 and beta-HSP90, and it forms dimers which are crucial species for its biological activity. PAGE, ESI-MS and MALDI-MS were used to study...... forms of native pig brain alpha-HSP90, with the diphosphorylated form being the most abundant. For the beta isoform, the di-phosphorylated was also the most abundant. MALDI mass spectra of HSP90 samples after chemical cross-linking showed a high percentage of alpha-alpha homodimers. In addition...

  8. Expression of HSP70 in Peripheral Lymphocytes of the Patients with Allergic Rhinitis

    Institute of Scientific and Technical Information of China (English)

    刘立思; 肖成峰; 张明; 程磊; 王鄂芬; 邬堂春


    The expression levels of heat shock protein 70 (HSP70) from peripheral lymphocytes ofthe patients with allergic rhinitis (AR) and the clinical implication were investigated. In the morn-ing, 3 ml of fasting venous blood was taken out. The lymphocytes were isolated by using Ficoll-Hypaque and the expression of HSP70 in the lymphocytes was detected by using Western blot. Inthe AR patients the HSP70 level (41.49± 15.77 integrated optical density, IOD) were significantlyhigher than that in the control group (23.89±10. 13 IOD, P<0.05). Western blot demonstratedthat HSP70 bands in AR patients were more intensive than those in the control group. It was con-cluded that the elevated HSP70 level in peripheral lymphocytes of the AR patients might contributeto the development of AR.

  9. Dynamic Analysis of the Expression of HSP70 during Experimental Tooth Movement in Rats

    Institute of Scientific and Technical Information of China (English)


    In this study, the expression of HSP70 during experimental tooth movement was dynamically observed and the relationship between HSP70 and orthodontic periodontal tissue remodeling were observed. The orthodontic appliance was placed between the right maxillary first molar and maxillary central incisors of adult SD rats to establish a rat molar movement model. Immunohistochemistry was performed 1, 3, 5, 7 and 14 day(s) after orthodontic force application to observe the expression and localization of inducible HSP70. The expression of HSP70 was strongly positive in the early stage of the tooth movement, became gradually less positive, and was weakly positive in the restoration stage. There was difference in staining pattern between different parts of PDL during the same period. These results suggest that the expression of HSP70 and difference in staining pattern among different parts of PDL during orthodontic tooth movement in rats may be implicated in stress response and remodeling of periodontal tissue.

  10. Crosstalk between p38, Hsp25 and Akt in spinal motor neurons after sciatic nerve injury (United States)

    Murashov, A. K.; Ul Haq, I.; Hill, C.; Park, E.; Smith, M.; Wang, X.; Wang, X.; Goldberg, D. J.; Wolgemuth, D. J.


    The p38 stress-activated protein kinase pathway is involved in regulation of phosphorylation of Hsp25, which in turn regulates actin filament dynamic in non-neuronal cells. We report that p38, Hsp25 and Akt signaling pathways were specifically activated in spinal motor neurons after sciatic nerve axotomy. The activation of the p38 kinase was required for induction of Hsp25 expression. Furthermore, Hsp25 formed a complex with Akt, a member of PI-3 kinase pathway that prevents neuronal cell death. Together, our observations implicate Hsp25 as a central player in a complex system of signaling that may both promote regeneration of nerve fibers and prevent neuronal cell death in the injured spinal cord.

  11. Adult Heat Tolerance Variation in Drosophila melanogaster is Not Related to Hsp70 Expression

    DEFF Research Database (Denmark)

    Jensen, Louise Toft; Cockerell, Fiona Elizabeth; Kristensen, Torsten Nygaard;


    Expression of heat-inducible Hsp70 is considered closely linked to thermotolerance in Drosophila melanogaster and other ectotherms. However, intra-specific variation of Hsp70 expression levels and its relationship to heat resistance has only been investigated in a few studies. Although...... in adult heat tolerance. Therefore, although Hsp70 expression is a major component of the cellular heat stress response, its influence on intra-specific heat tolerance variation may be life-stage specific. J. Exp. Zool. 313A:35-44, 2010. © 2009 Wiley-Liss, Inc...... in Drosophila larvae Hsp70 expression may be a key determinant of heat tolerance, the evidence for this in adults is equivocal. We therefore examined heat-induced Hsp70 expression and several measurements of adult heat tolerance in three independent collections of D. melanogaster, measured in three laboratories...

  12. Paralog-selective Hsp90 inhibitors define tumor-specific regulation of Her2 (United States)

    Patel, Pallav D.; Yan, Pengrong; Seidler, Paul M.; Patel, Hardik J.; Sun, Weilin; Yang, Chenghua; Que, Nanette S.; Taldone, Tony; Finotti, Paola; Stephani, Ralph A.; Gewirth, Daniel T.; Chiosis, Gabriela


    Although the Hsp90 chaperone family, comprised in humans of four paralogs, Hsp90α, Hsp90β, Grp94 and Trap-1, has important roles in malignancy, the contribution of each paralog to the cancer phenotype is poorly understood. This is in large part because reagents to study paralog-specific functions in cancer cells have been unavailable. Here we combine compound library screening with structural and computational analyses to identify purine-based chemical tools that are specific for Hsp90 paralogs. We show that Grp94 selectivity is due to the insertion of these compounds into a new allosteric pocket. We use these tools to demonstrate that cancer cells use individual Hsp90 paralogs to regulate a client protein in a tumor-specific manner and in response to proteome alterations. Finally, we provide new mechanistic evidence explaining why selective Grp94 inhibition is particularly efficacious in certain breast cancers. PMID:23995768

  13. Thermoluminescent response of dosemeters TLD-100 exposed to distinct baker protocols, irradiation and reading; Respuesta termoluminiscente de dosimetros TLD-100 sujetos a distintos protocolos de horneado, irradiacion y lectura

    Energy Technology Data Exchange (ETDEWEB)

    Estrada E, D


    The interest that motivated the realization of this work is the opposing discrepancy among the values of the efficiency thermoluminescent of heavy charged particles (PCP) regarding gamma radiation ({eta}{sub PCP}, {gamma}) reported in different works and gathered recently for Horowitz (Ho 01). The measures of relative thermoluminescent efficiency have importance in relation to the dosimetry associated to medical therapy with heavy charged particle. The measurement of {eta}{sub PCP}, {gamma} depends of multiple experimental factors. To understand this dependence it was quantified the effect of using different procedures experimental in the estimate of the relative thermoluminescent efficiency of dosemeters TLD-100 (LiF: Ti; Mg) for protons of 3 MeV regarding gamma rays of {sup 60}Co ({eta}{sub PCP}, {gamma}), varying the lot, the presentation, the one baked and the team reader. When was used the same global procedure values they were obtained for ({eta}{sub PCP}, {gamma}) inside the interval [0.30, 0.58] for the peak 5 and [0.35, 0.76] for the total thermoluminescent signal (Tl). At the use different equipment, baked and lot are obtained maximum differences of 30%, 12% and 6% respectively in the values of ({eta}{sub PCP}, {gamma}) for the peak 5, and 25%, 28% and 7% for the total Tl signal. The changes more significant of 36% and 44% for peak 5 and total signal respectively are obtained when changing the presentation of the dosemeter. As a complementary investigation it was studied the distribution of the Tl signal of each peak in the deconvolution of the curve of brightness with regard to the total signal in exposed dosemeters to high fluence of protons (1x10{sup 10} p/cm{sup 2}) varying the experimental protocol. It was found that the distribution of the Tl signal has a difference of the order of 5% when changing the used reading team. It was observed that the contribution of the peak 5 regarding the total signal are preserved when changing baked and that it

  14. BioAir: Bio-Inspired Airborne Infrastructure Reconfiguration (United States)


    UAV ) in order to autonomously form and maintain a dynamic communication network. This system draws upon inspirations from biological cell...presents the BioAIR system for autonomous communication , command and control of a swarm of UAVs . The idea is to form a communication network comprised of...deployment, yet maintaining communications should not interfere with the primary tasks of these entities. The BioAIR system was developed to coordinate

  15. Novel arrangement and comparative analysis of hsp90 family genes in three thermotolerant species of Stratiomyidae (Diptera). (United States)

    Astakhova, L N; Zatsepina, O G; Przhiboro, A A; Evgen'ev, M B; Garbuz, D G


    The heat shock proteins belonging to the Hsp90 family (Hsp83 in Diptera) play a crucial role in the protection of cells due to their chaperoning functions. We sequenced hsp90 genes from three species of the family Stratiomyidae (Diptera) living in thermally different habitats and characterized by extraordinarily high thermotolerance. The sequence variation and structure of the hsp90 family genes were compared with previously described features of hsp70 copies isolated from the same species. Two functional hsp83 genes were found in the species studied, that are arranged in tandem orientation at least in one of them. This organization was not previously described. Stratiomyidae hsp83 genes share a high level of identity with hsp83 of Drosophila, and the deduced protein possesses five conserved amino acid sequence motifs characteristic of the Hsp90 family as well as the C-terminus MEEVD sequence characteristic of the cytosolic isoform. A comparison of the hsp83 promoters of two Stratiomyidae species from thermally contrasting habitats demonstrated that while both species contain canonical heat shock elements in the same position, only one of the species contains functional GAF-binding elements. Our data indicate that in the same species, hsp83 family genes show a higher evolution rate than the hsp70 family.

  16. Xaa-Arg-Gly triplets in the collagen triple helix are dominant binding sites for the molecular chaperone HSP47. (United States)

    Koide, Takaki; Takahara, Yoshifumi; Asada, Shinichi; Nagata, Kazuhiro


    HSP47 is an essential procollagen-specific molecular chaperone that resides in the endoplasmic reticulum of procollagen-producing cells. Recent advances have revealed that HSP47 recognizes the (Pro-Pro-Gly)(n) sequence but not (Pro-Hyp-Gly)(n) and that HSP47 recognizes the triple-helical conformation. In this study, to better understand the substrate recognition by HSP47, we synthesized various collagen model peptides and examined their interaction with HSP47 in vitro. We found that the Pro-Arg-Gly triplet forms an HSP47-binding site. The HSP47 binding was observed only when Arg residues were incorporated in the Yaa positions of the Xaa-Yaa-Gly triplets. Amino acids in the Xaa position did not largely affect the interaction. The recognition of the Arg residue by HSP47 was specific to its side-chain structure because replacement of the Arg residue by other basic amino acids decreased the affinity to HSP47. The significance of Arg residues in HSP47 binding was further confirmed by using residue-specific chemical modification of types I and III collagen. Our results demonstrate that Xaa-Arg-Gly sequences in the triple-helical procollagen molecule are dominant binding sites for HSP47 and enable us to predict HSP47-binding sites in homotrimeric procollagen molecules.

  17. The molecular chaperone Hsp70 activates protein phosphatase 5 (PP5) by binding the tetratricopeptide repeat (TPR) domain. (United States)

    Connarn, Jamie N; Assimon, Victoria A; Reed, Rebecca A; Tse, Eric; Southworth, Daniel R; Zuiderweg, Erik R P; Gestwicki, Jason E; Sun, Duxin


    Protein phosphatase 5 (PP5) is auto-inhibited by intramolecular interactions with its tetratricopeptide repeat (TPR) domain. Hsp90 has been shown to bind PP5 to activate its phosphatase activity. However, the functional implications of binding Hsp70 to PP5 are not yet clear. In this study, we find that both Hsp90 and Hsp70 bind to PP5 using a luciferase fragment complementation assay. A fluorescence polarization assay shows that Hsp90 (MEEVD motif) binds to the TPR domain of PP5 almost 3-fold higher affinity than Hsp70 (IEEVD motif). However, Hsp70 binding to PP5 stimulates higher phosphatase activity of PP5 than the binding of Hsp90. We find that PP5 forms a stable 1:1 complex with Hsp70, but the interaction appears asymmetric with Hsp90, with one PP5 binding the dimer. Solution NMR studies reveal that Hsc70 and PP5 proteins are dynamically independent in complex, tethered by a disordered region that connects the Hsc70 core and the IEEVD-TPR contact area. This tethered binding is expected to allow PP5 to carry out multi-site dephosphorylation of Hsp70-bound clients with a range of sizes and shapes. Together, these results demonstrate that Hsp70 recruits PP5 and activates its phosphatase activity which suggests dual roles for PP5 that might link chaperone systems with signaling pathways in cancer and development.

  18. Navigating the Bio-Politics of Childhood (United States)

    Lee, Nick; Motzkau, Johanna


    Childhood research has long shared a bio-political terrain with state agencies in which children figure primarily as "human futures". In the 20th century bio-social dualism helped to make that terrain navigable by researchers, but, as life processes increasingly become key sites of bio-political action, bio-social dualism is becoming…

  19. Induction of premature senescence by hsp90 inhibition in small cell lung cancer.

    Directory of Open Access Journals (Sweden)

    Ian J Restall

    Full Text Available BACKGROUND: The molecular chaperone Hsp90 is a promising new target in cancer therapy and selective Hsp90 inhibitors are currently in clinical trials. Previously these inhibitors have been reported to induce either cell cycle arrest or cell death in cancer cells. Whether the cell cycle arrest is reversible or irreversible has not generally been assessed. Here we have examined in detail the cell cycle arrest and cell death responses of human small cell lung cancer cell lines to Hsp90 inhibition. METHODOLOGY/PRINCIPAL FINDINGS: In MTT assays, small cell lung cancer cells showed a biphasic response to the Hsp90 inhibitors geldanamycin and radicicol, with low concentrations causing proliferation arrest and high concentrations causing cell death. Assessment of Hsp90 intracellular activity using loss of client protein expression showed that geldanamycin concentrations that inhibited Hsp90 correlated closely with those causing proliferation arrest but not cell death. The proliferation arrest induced by low concentrations of geldanamycin was not reversed for a period of over thirty days following drug removal and showed features of senescence. Rare populations of variant small cell lung cancer cells could be isolated that had additional genetic alterations and no longer underwent irreversible proliferation arrest in response to Hsp90 inhibitors. CONCLUSIONS/SIGNIFICANCE: We conclude that: (1 Hsp90 inhibition primarily induces premature senescence, rather than cell death, in small cell lung cancer cells; (2 small cell lung cancer cells can bypass this senescence through further genetic alterations; (3 Hsp90 inhibitor-induced cell death in small cell lung cancer cells is due to inhibition of a target other than cytosolic Hsp90. These results have implications with regard to how these inhibitors will behave in clinical trials and for the design of future inhibitors in this class.

  20. Ste20-like kinase, SLK, activates the heat shock factor 1 - Hsp70 pathway. (United States)

    Cybulsky, Andrey V; Guillemette, Julie; Papillon, Joan


    Expression and activation of SLK increases during renal ischemia-reperfusion injury. When highly expressed, SLK signals via c-Jun N-terminal kinase and p38 to induce apoptosis, and it exacerbates apoptosis induced by ischemia-reperfusion injury. Overexpression of SLK in glomerular epithelial cells (GECs)/podocytes in vivo induces injury and proteinuria. In response to various stresses, cells enhance expression of chaperones or heat shock proteins (e.g. Hsp70), which are involved in the folding and maturation of newly synthesized proteins, and can refold denatured or misfolded proteins. We address the interaction of SLK with the heat shock factor 1 (HSF1)-Hsp70 pathway. Increased expression of SLK in GECs (following transfection) induced HSF1 transcriptional activity. Moreover, HSF1 transcriptional activity was increased by in vitro ischemia-reperfusion injury (chemical anoxia/recovery) and heat shock, and in both instances was amplified further by SLK overexpression. HSF1 binds to promoters of target genes, such as Hsp70 and induces their transcription. By analogy to HSF1, SLK stimulated Hsp70 expression. Hsp70 was also enhanced by anoxia/recovery and was further amplified by SLK overexpression. Induction of HSF1 and Hsp70 was dependent on the kinase activity of SLK, and was mediated via polo-like kinase-1. Transfection of constitutively active HSF1 enhanced Hsp70 expression and inhibited SLK-induced apoptosis. Conversely, the proapoptotic action of SLK was augmented by HSF1 shRNA, or the Hsp70 inhibitor, pifithrin-μ. In conclusion, increased expression/activity of SLK activates the HSF1-Hsp70 pathway. Hsp70 attenuates the primary proapoptotic effect of SLK. Modulation of chaperone expression may potentially be harnessed as cytoprotective therapy in renal cell injury.

  1. Chicken HSP70 DNA vaccine inhibits tumor growth in a canine cancer model. (United States)

    Yu, Wen-Ying; Chuang, Tien-Fu; Guichard, Cécile; El-Garch, Hanane; Tierny, Dominique; Laio, Albert Taiching; Lin, Ching-Si; Chiou, Kuo-Hao; Tsai, Cheng-Long; Liu, Chen-Hsuan; Li, Wen-Chiuan; Fischer, Laurent; Chu, Rea-Min


    Immunization with xenogeneic DNA is a promising cancer treatment to overcome tolerance to self-antigens. Heat shock protein 70 (HSP70) is over-expressed in various kinds of tumors and is believed to be involved in tumor progression. This study tested a xenogeneic chicken HSP70 (chHSP70) DNA vaccine in an experimental canine transmissible venereal tumor (CTVT) model. Three vaccination strategies were compared: the first (PE) was designed to evaluate the prophylactic efficacy of chHSP70 DNA vaccination by delivering the vaccine before tumor inoculation in a prime boost setting, the second (T) was designed to evaluate the therapeutic efficacy of the same prime boost vaccine by vaccinating the dogs after tumor inoculation; the third (PT) was similar to the first strategy (PE), with the exception that the electroporation booster injection was replaced with a transdermal needle-free injection. Tumor growth was notably inhibited only in the PE dogs, in which the vaccination program triggered tumor regression significantly sooner than in control dogs (NT). The CD4(+) subpopulation of tumor-infiltrating lymphocytes and canine HSP70 (caHSP70)-specific IFN-γ-secreting lymphocytes were significantly increased during tumor regression in the PE dogs as compared to control dogs, demonstrating that specific tolerance to caHSP70 has been overcome. In contrast, no benefit of the therapeutic strategy (T) could be noticed and the (PT) strategy only led to partial control of tumor growth. In summary, antitumor prophylactic activity was demonstrated using the chHSP70 DNA vaccine including a boost via electroporation. Our data stressed the importance of DNA electroporation as a booster to get the full benefit of DNA vaccination but also of cancer immunotherapy initiation as early as possible. Xenogeneic chHSP70 DNA vaccination including an electroporation boost is a potential vaccine to HSP70-expressing tumors, although further research is still required to better understand true

  2. E. coli chaperones DnaK, Hsp33 and Spy inhibit bacterial functional amyloid assembly. (United States)

    Evans, Margery L; Schmidt, Jens C; Ilbert, Marianne; Doyle, Shannon M; Quan, Shu; Bardwell, James C A; Jakob, Ursula; Wickner, Sue; Chapman, Matthew R


    Amyloid formation is an ordered aggregation process, where β-sheet rich polymers are assembled from unstructured or partially folded monomers. We examined how two Escherichia coli cytosolic chaperones, DnaK and Hsp33, and a more recently characterized periplasmic chaperone, Spy, modulate the aggregation of a functional amyloid protein, CsgA. We found that DnaK, the Hsp70 homologue in E. coli, and Hsp33, a redox-regulated holdase, potently inhibited CsgA amyloidogenesis. The Hsp33 anti-amyloidogenesis activity was oxidation dependent, as oxidized Hsp33 was significantly more efficient than reduced Hsp33 at preventing CsgA aggregation. When soluble CsgA was seeded with preformed amyloid fibers, neither Hsp33 nor DnaK were able to efficiently prevent soluble CsgA from adopting the amyloid conformation. Moreover, both DnaK and Hsp33 increased the time that CsgA was reactive with the amyloid oligomer conformation-specific A11 antibody. Since CsgA must also pass through the periplasm during secretion, we assessed the ability of the periplasmic chaperone Spy to inhibit CsgA polymerization. Like DnaK and Hsp33, Spy also inhibited CsgA polymerization in vitro. Overexpression of Spy resulted in increased chaperone activity in periplasmic extracts and in reduced curli biogenesis in vivo. We propose that DnaK, Hsp33 and Spy exert their effects during the nucleation stages of CsgA fibrillation. Thus, both housekeeping and stress induced cytosolic and periplasmic chaperones may be involved in discouraging premature CsgA interactions during curli biogenesis.

  3. Administration of Mycobacterium leprae rHsp65 aggravates experimental autoimmune uveitis in mice.

    Directory of Open Access Journals (Sweden)

    Eliana B Marengo

    Full Text Available The 60 kDa heat shock protein family, Hsp60, constitutes an abundant and highly conserved class of molecules that are highly expressed in chronic-inflammatory and autoimmune processes. Experimental autoimmune uveitis [EAU] is a T cell mediated intraocular inflammatory disease that resembles human uveitis. Mycobacterial and homologous Hsp60 peptides induces uveitis in rats, however their participation in aggravating the disease is poorly known. We here evaluate the effects of the Mycobacterium leprae Hsp65 in the development/progression of EAU and the autoimmune response against the eye through the induction of the endogenous disequilibrium by enhancing the entropy of the immunobiological system with the addition of homologous Hsp. B10.RIII mice were immunized subcutaneously with interphotoreceptor retinoid-binding protein [IRBP], followed by intraperitoneally inoculation of M. leprae recombinant Hsp65 [rHsp65]. We evaluated the proliferative response, cytokine production and the percentage of CD4(+IL-17(+, CD4(+IFN-gamma(+ and CD4(+Foxp3(+ cells ex vivo, by flow cytometry. Disease severity was determined by eye histological examination and serum levels of anti-IRBP and anti-Hsp60/65 measured by ELISA. EAU scores increased in the Hsp65 group and were associated with an expansion of CD4(+IFN-gamma(+ and CD4(+IL-17(+ T cells, corroborating with higher levels of IFN-gamma. Our data indicate that rHsp65 is one of the managers with a significant impact over the immune response during autoimmunity, skewing it to a pathogenic state, promoting both Th1 and Th17 commitment. It seems comprehensible that the specificity and primary function of Hsp60 molecules can be considered as a potential pathogenic factor acting as a whistleblower announcing chronic-inflammatory diseases progression.

  4. Differences in the curing of [PSI+] prion by various methods of Hsp104 inactivation.

    Directory of Open Access Journals (Sweden)

    Yang-Nim Park

    Full Text Available [PSI(+] yeast, containing the misfolded amyloid conformation of Sup35 prion, is cured by inactivation of Hsp104. There has been controversy as to whether inactivation of Hsp104 by guanidine treatment or by overexpression of the dominant negative Hsp104 mutant, Hsp104-2KT, cures [PSI(+] by the same mechanism- inhibition of the severing of the prion seeds. Using live cell imaging of Sup35-GFP, overexpression of Hsp104-2KT caused the foci to increase in size, then decrease in number, and finally disappear when the cells were cured, similar to that observed in cells cured by depletion of Hsp104. In contrast, guanidine initially caused an increase in foci size but then the foci disappeared before the cells were cured. By starving the yeast to make the foci visible in cells grown with guanidine, the number of cells with foci was found to correlate exactly with the number of [PSI(+] cells, regardless of the curing method. Therefore, the fluorescent foci are the prion seeds required for maintenance of [PSI(+] and inactivation of Hsp104 cures [PSI(+] by preventing severing of the prion seeds. During curing with guanidine, the reduction in seed size is an Hsp104-dependent effect that cannot be explained by limited severing of the seeds. Instead, in the presence of guanidine, Hsp104 retains an activity that trims or reduces the size of the prion seeds by releasing Sup35 molecules that are unable to form new prion seeds. This Hsp104 activity may also occur in propagating yeast.

  5. Expression analysis of HSP70 in the testis of Octopus tankahkeei under thermal stress. (United States)

    Long, Ling-Li; Han, Ying-Li; Sheng, Zhang; Du, Chen; Wang, You-Fa; Zhu, Jun-Quan


    The gene encoding heat shock protein 70 (HSP70) was identified in Octopus tankahkeei by homologous cloning and rapid amplification of cDNA ends (RACE). The full-length cDNA (2471 bp) consists of a 5'-untranslated region (UTR) (89 bp), a 3'-UTR (426 bp), and an open reading frame (1956 bp) that encodes 651 amino acid residues with a predicted molecular mass of 71.8 kDa and an isoelectric point of 5.34. Based on the amino acid sequence analysis and multiple sequence alignment, this cDNA is a member of cytoplasmic hsp70 subfamily of the hsp70 family and was designated as ot-hsp70. Tissue expression analysis showed that HSP70 expression is highest in the testes when all examined organs were compared. Immunohistochemistry analysis, together with hematoxylin-eosin staining, revealed that the HSP70 protein was expressed in all spermatogenic cells, but not in fibroblasts. In addition, O. tankahkeei were heat challenged by exposure to 32 °C seawater for 2 h, then returned to 13 °C for various recovery time (0-24 h). Relative expression of ot-hsp70 mRNA in the testes was measured at different time points post-challenge by quantitative real-time PCR. A clear time-dependent mRNA expression of ot-hsp70 after thermal stress indicates that the HSP70 gene is inducible. Ultrastructural changes of the heat-stressed testis were observed by transmission electron microscopy. We suggest that HSP70 plays an important role in spermatogenesis and testis protection against thermal stress in O. tankahkeei.

  6. Hsp70 vaccination-induced primary immune responses in efferent lymph of the draining lymph node. (United States)

    Vrieling, Manouk; Santema, Wiebren; Vordermeier, Martin; Rutten, Victor; Koets, Ad


    Bovine paratuberculosis is a highly prevalent chronic infection of the small intestine in cattle, caused by Mycobacterium avium subspecies paratuberculosis (MAP). In earlier studies we showed the protective effect of Hsp70/DDA subunit vaccination against paratuberculosis. In the current study we set out to measure primary immune responses generated at the site of Hsp70 vaccination. Lymph vessel cannulation was performed to obtain efferent lymph from the prescapular lymph node draining the neck area where the vaccine was applied. Hsp70 vaccination induced a significant increase of CD21(+) B cells in efferent lymph, accounting for up to 40% of efferent cells post-vaccination. Proliferation (Ki67(+)) within the CD21(+) B cell and CD4(+) T cell populations peaked between day 3 and day 5 post-vaccination. From day 7, Hsp70-specific antibody secreting cells (ASCs) could be detected in efferent lymph. Hsp70-specific antibodies, mainly of the IgG1 isotype, were also detected from this time point onwards. However, post-vaccination IFN-γ production in efferent lymph was non-sustained. In conclusion, Hsp70-vaccination induces only limited Th1 type immune responsiveness as reflected in efferent lymph draining the vaccination site. This is in line with our previous observations in peripheral blood. The main primary immunological outcome of the Hsp70/DDA subunit vaccination is B cell activation and abundant Hsp70-specific IgG1 production. This warrants the question whether Hsp70-specific antibodies contribute to the observed protective effect of Hsp70 vaccination in calves.

  7. NIH NeuroBioBank (United States)

    Federal Laboratory Consortium — The NIH NeuroBioBank (NBB), supported by the National Institute of Mental Health, the National Institute of Neurological Disorders and Stroke, and the Eunice Kennedy...

  8. Thermoluminescent and optical processes in alkaline halogenides dosemeters contaminated with Europium; Procesos opticos y termoluminiscentes en dosimetros de halogenuros alcalinos contaminados con Europio

    Energy Technology Data Exchange (ETDEWEB)

    Barboza F, M.; Melendrez, R.; Castaneda, B.; Pedroza M, M.; Chernov, V.; Perez S, R.; Aceves, R. [Centro de Investigacion en Fisica, Universidad de Sonora, A.P. 5-088, 83190 Hermosillo, Sonora (Mexico)


    Recent research results are presented about the properties of the optical processes of photo transferred thermoluminescence (TLFT), optical whitening (BO), thermoluminescence induced by light (TLL) and its effect in the thermoluminescent curve (Tl) produced by ionizing and non-ionizing radiation. The systematic analysis of all these processes, acquires a singular importance due that actually the alkaline halogenide crystals are object of intense investigations which analyse their potential applications as detectors and radiation dosemeters through stimulated optical luminescence techniques or thermoluminescence. The obtained data show that the Tl curve of material with this nature can be enormously affected by exposure of phosphorus to the environmental light or UV. This is in part due to liberation processes of charge bearers are shouted and makes a subsequent trapping in less temperature traps; at the same time that induce changes in the intensity of determined Tl bands. Additionally, also it is observed that mentioned phenomena are related as with wavelength of incident light as of the illumination time. Finally, the obtained information allows to conclude that although the illumination effect is extremely complex, it is associated and can be explained mainly with phenomena that implicate the electrons excitation trapped in form of F centers and trapping mechanisms or radioactive and non-radioactive recombination. (Author)

  9. Phosphorylation of an envelope-associated Hsp70 homolog in amyloplasts isolated from cultured cells of sycamore (Acer pseudoplatanus L.). (United States)

    Checa, S K; Viale, A M


    The presence of Hsp70 and Hsp60 molecular chaperones in amyloplasts isolated from cultured sycamore cells was analyzed by immunoblotting. Hsp70 homologs were located in both amyloplast envelope and stromal fractions, but no Hsp60 homologs were detected in any of the different suborganellar fractions. Incubation of whole amyloplasts or their envelope fraction with Mg2+ gamma-32P-ATP resulted in a rapid phosphorylation of the envelope-associated Hsp70 homolog, which constitutes a major target of phosphorylation in these plastids.

  10. Synthesis and evaluation of cell-permeable biotinylated PU-H71 derivatives as tumor Hsp90 probes

    Directory of Open Access Journals (Sweden)

    Tony Taldone


    Full Text Available The attachment of biotin to a small molecule provides a powerful tool in biology. Here, we present a systematic approach to identify biotinylated analogues of the Hsp90 inhibitor PU-H71 that are capable of permeating cell membranes so as to enable the investigation of Hsp90 complexes in live cells. The identified derivative 2g can isolate Hsp90 through affinity purification and, as we show, represents a unique and useful tool to probe tumor Hsp90 biology in live cells by affinity capture, flow cytometry and confocal microscopy. To our knowledge, 2g is the only reported biotinylated Hsp90 probe to have such combined characteristics.

  11. Synthesis and evaluation of cell-permeable biotinylated PU-H71 derivatives as tumor Hsp90 probes (United States)

    DaGama Gomes, Erica M; Riolo, Matthew; Patel, Hardik J; Alonso-Sabadell, Raul; Zatorska, Danuta; Patel, Maulik R; Kishinevsky, Sarah


    Summary The attachment of biotin to a small molecule provides a powerful tool in biology. Here, we present a systematic approach to identify biotinylated analogues of the Hsp90 inhibitor PU-H71 that are capable of permeating cell membranes so as to enable the investigation of Hsp90 complexes in live cells. The identified derivative 2g can isolate Hsp90 through affinity purification and, as we show, represents a unique and useful tool to probe tumor Hsp90 biology in live cells by affinity capture, flow cytometry and confocal microscopy. To our knowledge, 2g is the only reported biotinylated Hsp90 probe to have such combined characteristics. PMID:23616796

  12. Mensuration of equivalent dose with personal dosemeters and instruments of radiological protection in the new operative magnitudes ICRU, for external fields of beta radiation. Part I. Study of the homogeneity of the response personal dosemeters leaves (cards G-1, TLD-100), in radiation fields of Co{sub 60}; Medicion de dosis equivalente con dosimetros personales e instrumentos de proteccion radiologica en las nuevas magnitudes operativas ICRU, para campos de radiacion beta externos. Parte I. Estudio de la homogeneidad de la respuesta dosimetros personales (tarjetas G-1, TLD-100), en campos de radiacion de Co{sub 60}

    Energy Technology Data Exchange (ETDEWEB)

    Alvarez R, J.T. [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico)


    A sample of 40 composed personal dosemeters by cards model: G-l, (each card is made up of two TLD-100 crystals encapsulated in teflon), Harshaw trademark; those personal dosemeters present a free window and another with a filter of A1 of 171.7 mg cm{sup -2} of mass thickness.The objective of the work is to select of this sample of 40 personal dosemeters a population with the same stocking and standard deviation. The technique used is that of comparison of stockings, (ANOVA; Variance Analysis, when samples of the same one were had size; and/or GLM, Widespread Lineal Models, when the samples were of different size), by means of the use of those Duncan statistics, SNK, Tukey, Gabriel; the results are validated proving the kindness of adjustment of the experimental data to a Normal distribution by means of the Shapiro-Wilks statistics.The experimental design used consists on a test of two vias: a via is the variable card with two levels, (crystal 1 and 2), the other via is the variable irradiation position with four levels, (LS=left superior, SR= right superior, LI= left Inferior, IR = right inferior). The irradiations carried out in blocks of four personal dosemeters in a gamma radiation beam range of Cobalt 60; carrying out three repetitions of the design. With object of proving the homogeneity of the filter of A1 in those personal dosemeters the experimental design was executed for those cards without personal dosemeters.They were also carried out tests of stockings to the readings of bottom and sensibility of the reader equipment, (Harshaw, model marks 2271), certain that doesn{sup t} exist differences for sequence of reading, but if in the stockings of the sensibility, (they were 4 different populations). The responses of the dosemeters were corrected subtracting him the reading correspondence of bottom and by sensibility of the reader equipment before subjecting them to the tests of stockings mentioned. Of the results of the tests of stockings for the cards with

  13. Research Advances of HSP27's Role in Cardiovascular Diseases%HSP27在心血管疾病中作用的研究进展

    Institute of Scientific and Technical Information of China (English)



    热休克蛋白27(HSP27)是小分子HSP家族中的重要成员,细胞在外界自由基、热、缺血和毒性物质等刺激下可诱导其产生,在各种肿瘤和正常组织中特别是心脏组织中呈高表达.在对抗细心肌凋亡、抗缺血损伤、抗炎性反应中HSP27都有着积极的作用,对血管平滑肌细胞的增殖、迁移和动脉粥样硬化的发生和发展过程都有调控作用.HSP27可能成为治疗心血管疾病的新途径.%HSP27 is an important member of small molecular weight heat shock protein( HSP ) family, the external stimulation of free radicals, heat, ischemic and toxic substances can induce the generation of HSP27 in the cell, presenting high expression in a variety of tumors and normal tissues, with especially high expression in cardiac tissue, which have a positive effect in the anti-apoptosis, anti-ischemia injury and anti-inflammatory. It has a regulatory role in the vascular smooth muscle cell proliferation and migration and the occurrence and development of atherosclerosis. HSP27 may be a new therapeutic strategy of cardiovascular diseases.

  14. Induction of a heat shock response (HSP 72) in rat embryos exposed to selected chemical teratogens. (United States)

    Mirkes, P E; Doggett, B; Cornel, L


    A monoclonal antibody to the 72 kD heat shock protein (HSP 72), Western blot analysis and 2-D gel electrophoresis/autoradiography were used to determine whether selected chemical teratogens induced the synthesis and accumulation of HSP 72 in postimplantation rat embryos exposed in vitro. The chemical teratogens studied include N-Acetoxy-2-acetylaminofluorene (N-Ac-AAF), cadmium chloride (CAD), cyclophosphamide (CP), sodium arsenite (AS), and sodium salicylate (SAL). Exposures to test chemicals were selected that produced obvious embryotoxicity characterized by abnormal development and growth retardation. Of the five chemical teratogens studied, AS and SAL induced the synthesis and accumulation of HSP 72 in day 10 rat embryos. The kinetics of HSP 72 accumulation, however, differed between AS- and SAL-treated embryos. Maximal levels of HSP 72 were observed 24 hours after AS exposure and 10 hours after SAL exposure. N-Ac-AAF, CD, and CP induced obvious embryotoxicity; however, none of these chemical teratogens induced HSP 72 at any of the timepoints assayed. Although only a small sample of chemical teratogens was studied, our results suggest that the heat shock response, characterized by the synthesis and accumulation of HSP 72, is not a general biomarker for chemical teratogens.

  15. Molecular cloning of HSP70 in Mycoplasma ovipneumoniae and comparison with that of other mycoplasmas. (United States)

    Li, M; Ma, C J; Liu, X M; Zhao, D; Xu, Q C; Wang, Y J


    Mycoplasma ovipneumoniae, a bacterial species that specifically affects ovine and goat, is the cause of ovine infectious pleuropneumonia. We cloned, sequenced and analyzed heat shock protein 70 (HSP70) (dnaK) gene of M. ovipneumoniae. The full length open reading frame of the M. ovipneumoniae HSP70 gene consists of 1812 nucleotides, with a G+C content of 34.16%, encoding 604 amino acids. Comparative analysis with the HSP70 sequences of 15 Mycoplasma species revealed 59 to 87% DNA sequence identity, with an amino acid sequence identity range of 58 to 94%. M. ovipneumoniae and M. hyopneumoniae shared the highest DNA and amino acid sequence identity (87 and 94%, respectively). Based on phylogenetic analysis, both the DNA and amino acid identities of M. ovipneumoniae with other mycoplasmal HSP70 were correlated with the degree of relationship between the species. The C-terminus of the HSP70 was cloned into a bacterial expression vector and expressed in Escherichia coli cells. The recombinant C-terminal portion of HSP70 protein strongly reacted with convalescent sera from M. ovipneumoniae-infected sheep, based on an immunoblotting assay. This indicates that HSP70 is immunogenic in a natural M. ovipneumoniae infection and may be a relevant antigen for vaccine development.

  16. Localization of MRP-1 to the outer mitochondrial membrane by the chaperone protein HSP90β. (United States)

    Roundhill, Elizabeth; Turnbull, Doug; Burchill, Susan


    Overexpression of plasma membrane multidrug resistance-associated protein 1 (MRP-1) in Ewing's sarcoma (ES) predicts poor outcome. MRP-1 is also expressed in mitochondria, and we have examined the submitochondrial localization of MRP-1 and investigated the mechanism of MRP-1 transport and role of this organelle in the response to doxorubicin. The mitochondrial localization of MRP-1 was examined in ES cell lines by differential centrifugation and membrane solubilization by digitonin. Whether MRP-1 is chaperoned by heat shock proteins (HSPs) was investigated by immunoprecipitation, immunofluorescence microscopy, and HSP knockout using small hairpin RNA and inhibitors (apoptozole, 17-AAG, and NVPAUY). The effect of disrupting mitochondrial MRP-1-dependent efflux activity on the cytotoxic effect of doxorubicin was investigated by counting viable cell number. Mitochondrial MRP-1 is glycosylated and localized to the outer mitochondrial membrane, where it is coexpressed with HSP90. MRP-1 binds to both HSP90 and HSP70, although only inhibition of HSP90β decreases expression of MRP-1 in the mitochondria. Disruption of mitochondrial MRP-1-dependent efflux significantly increases the cytotoxic effect of doxorubicin (combination index, MRP-1 is expressed in the outer mitochondrial membrane and is a client protein of HSP90β, where it may play a role in the doxorubicin-induced resistance of ES.-Roundhill, E., Turnbull, D., Burchill, S. Localization of MRP-1 to the outer mitochondrial membrane by the chaperone protein HSP90β.

  17. Expression analysis of NOS family and HSP genes during thermal stress in goat ( Capra hircus) (United States)

    Yadav, Vijay Pratap; Dangi, Satyaveer Singh; Chouhan, Vikrant Singh; Gupta, Mahesh; Dangi, Saroj K.; Singh, Gyanendra; Maurya, Vijay Prakash; Kumar, Puneet; Sarkar, Mihir


    Approximately 50 genes other than heat shock protein (HSP) expression changes during thermal stress. These genes like nitric oxide synthase (NOS) need proper attention and investigation to find out their possible role in the adaptation to thermal stress in animals. So, the present study was undertaken to demonstrate the expressions of inducible form type II NOS (iNOS), endothelial type III NOS (eNOS), constitutively expressed enzyme NOS (cNOS), HSP70, and HSP90 in peripheral blood mononuclear cells (PBMCs) during different seasons in Barbari goats. Real-time polymerase chain reaction, western blot, and immunocytochemistry were applied to investigate messenger RNA (mRNA) expression, protein expression, and immunolocalization of examined factors. The mRNA and protein expressions of iNOS, eNOS, cNOS, HSP70, and HSP90 were significantly higher ( P < 0.05) during peak summer, and iNOS and eNOS expressions were also observed to be significantly higher ( P < 0.05) during peak winter season as compared with moderate season. The iNOS, eNOS, cNOS, HSP70, and HSP90 were mainly localized in plasma membrane and cytoplasm of PBMCs. To conclude, data generated in the present study indicate the possible involvement of the NOS family genes in amelioration of thermal stress so as to maintain cellular integrity and homeostasis in goats.

  18. Complexity of rice Hsp100 gene family: lessons from rice genome sequence data

    Indian Academy of Sciences (India)

    Gaurav Batra; Vineeta Singh Chauhan; Amanjot Singh; Neelam K Sarkar; Anil Grover


    Elucidation of genome sequence provides an excellent platform to understand detailed complexity of the various gene families. Hsp100 is an important family of chaperones in diverse living systems. There are eight putative gene loci encoding for Hsp100 proteins in Arabidopsis genome. In rice, two full-length Hsp100 cDNAs have been isolated and sequenced so far. Analysis of rice genomic sequence by in silico approach showed that two isolated rice Hsp100 cDNAs correspond to Os05g44340 and Os02g32520 genes in the rice genome database. There appears to be three additional proteins (encoded by Os03g31300, Os04g32560 and Os04g33210 gene loci) that are variably homologous to Os05g44340 and Os02g32520 throughout the entire amino acid sequence. The above five rice Hsp100 genes show significant similarities in the signature sequences known to be conserved among Hsp100 proteins. While Os05g44340 encodes cytoplasmic Hsp100 protein, those encoded by the other four genes are predicted to have chloroplast transit peptides.

  19. A preliminary study on functional domains of small heat shock protein Hsp16.3

    Institute of Scientific and Technical Information of China (English)

    CHANG Yong; LI Xuemei; RAO Zihe


    Hsp16.3, the small heat shock protein (sHSP) from Mycobacterium tuberculosis, was originally identified as an immunodominant antigen,which possesses three functional domains typical of sHSP family, namely the N-terminal hydrophobic region, α-crystallin domain and a short non-conserved C-terminal extension.To further understand the functional assignment of these independent regions, the three functional domains of Hsp16.3 were defined and the two N- or C-terminal truncated Hsp16.3 remnants were successfully cloned, expressed and purified.In the far and near circular dichroism analysis, the results showed that these remnants expressed similar secondary and tertiary structures to that of wild-type protein.During the reassembly of wild-type nonamer, the C-terminal truncated remnant could interact with the wild-type protein to form hetero-oligomers.When trypsin is used to digest the wild-type Hsp16.3, its α-crystallin domain could resist such degradation.Taken together, these results indicate that the stable secondary and tertiary structures of Hsp16.3 are mainly kept by its α-crystallin domain.

  20. HSF1-TPR interaction facilitates export of stress-induced HSP70 mRNA. (United States)

    Skaggs, Hollie S; Xing, Hongyan; Wilkerson, Donald C; Murphy, Lynea A; Hong, Yiling; Mayhew, Christopher N; Sarge, Kevin D


    Stress conditions inhibit mRNA export, but mRNAs encoding heat shock proteins continue to be efficiently exported from the nucleus during stress. How HSP mRNAs bypass this stress-associated export inhibition was not known. Here, we show that HSF1, the transcription factor that binds HSP promoters after stress to induce their transcription, interacts with the nuclear pore-associating TPR protein in a stress-responsive manner. TPR is brought into proximity of the HSP70 promoter after stress and preferentially associates with mRNAs transcribed from this promoter. Disruption of the HSF1-TPR interaction inhibits the export of mRNAs expressed from the HSP70 promoter, both endogenous HSP70 mRNA and a luciferase reporter mRNA. These results suggest that HSP mRNA export escapes stress inhibition via HSF1-mediated recruitment of the nuclear pore-associating protein TPR to HSP genes, thereby functionally connecting the first and last nuclear steps of the gene expression pathway, transcription and mRNA export.

  1. Conformational diversity in the TPR domain-mediated interaction of protein phosphatase 5 with Hsp90. (United States)

    Cliff, Matthew J; Harris, Richard; Barford, David; Ladbury, John E; Williams, Mark A


    Protein phosphatase 5 (Ppp5) is one of several proteins that bind to the Hsp90 chaperone via a tetratricopeptide repeat (TPR) domain. We report the solution structure of a complex of the TPR domain of Ppp5 with the C-terminal pentapeptide of Hsp90. This structure has the "two-carboxylate clamp" mechanism of peptide binding first seen in the Hop-TPR domain complexes with Hsp90 and Hsp70 peptides. However, NMR data reveal that the Ppp5 clamp is highly dynamic, and that there are multiple modes of peptide binding and mobility throughout the complex. Although this interaction is of very high affinity, relatively few persistent contacts are found between the peptide and the Ppp5-TPR domain, thus explaining its promiscuity in binding both Hsp70 and Hsp90 in vivo. We consider the possible implications of this dynamic structure for the mechanism of relief of autoinhibition in Ppp5 and for the mechanisms of TPR-mediated recognition of Hsp90 by other proteins.

  2. Client Proteins and Small Molecule Inhibitors Display Distinct Binding Preferences for Constitutive and Stress-Induced HSP90 Isoforms and Their Conformationally Restricted Mutants.

    Directory of Open Access Journals (Sweden)

    Thomas L Prince

    Full Text Available The two cytosolic/nuclear isoforms of the molecular chaperone HSP90, stress-inducible HSP90α and constitutively expressed HSP90β, fold, assemble and maintain the three-dimensional structure of numerous client proteins. Because many HSP90 clients are important in cancer, several HSP90 inhibitors have been evaluated in the clinic. However, little is known concerning possible unique isoform or conformational preferences of either individual HSP90 clients or inhibitors. In this report, we compare the relative interaction strength of both HSP90α and HSP90β with the transcription factors HSF1 and HIF1α, the kinases ERBB2 and MET, the E3-ubiquitin ligases KEAP1 and RHOBTB2, and the HSP90 inhibitors geldanamycin and ganetespib. We observed unexpected differences in relative client and drug preferences for the two HSP90 isoforms, with HSP90α binding each client protein with greater apparent affinity compared to HSP90β, while HSP90β bound each inhibitor with greater relative interaction strength compared to HSP90α. Stable HSP90 interaction was associated with reduced client activity. Using a defined set of HSP90 conformational mutants, we found that some clients interact strongly with a single, ATP-stabilized HSP90 conformation, only transiently populated during the dynamic HSP90 chaperone cycle, while other clients interact equally with multiple HSP90 conformations. These data suggest different functional requirements among HSP90 clientele that, for some clients, are likely to be ATP-independent. Lastly, the two inhibitors examined, although sharing the same binding site, were differentially able to access distinct HSP90 conformational states.

  3. Glutamine May Repress the Weak LPS and Enhance the Strong Heat Shock Induction of Monocyte and Lymphocyte HSP72 Proteins but May Not Modulate the HSP72 mRNA in Patients with Sepsis or Trauma

    Directory of Open Access Journals (Sweden)

    Efrossini Briassouli


    Full Text Available Objective. We assessed the lipopolysaccharide (LPS or heat shock (HS induction of heat shock protein-72 (HSP72 in peripheral blood mononuclear cells (PBMCs of patients with severe sepsis (SS or trauma-related systemic inflammatory response syndrome (SIRS, compared to healthy individuals (H; we also investigated any pre- or posttreatment modulating glutamine (Gln effect. Methods. SS (11, SIRS (10, and H (19 PBMCs were incubated with 1 μg/mL LPS or 43°HS. Gln 10 mM was either added 1 h before or 1 h after induction or was not added at all. We measured monocyte (m, lymphocyte (l, mRNA HSP72, HSP72 polymorphisms, interleukins (ILs, monocyte chemoattractant protein-1 (MCP-1, and cortisol levels. Results. Baseline lHSP72 was higher in SS p<0.03, and mHSP72 in SIRS p<0.02, compared to H. Only HS induced l/mHSP72/mRNA HSP72; LPS induced IL-6, IL-8, IL-10, and MCP-1. Induced mRNA was related to l/mHSP72, and was related negatively to cytokines. Intracellular l/mHSP72/HSP72 mRNA was related to serum ILs, not being influenced by cortisol, illness severity, and HSP72 polymorphisms. Gln did not induce mRNA in any group but modified l/mHSP72 after LPS/HS induction unpredictably. Conclusions. HSP72 mRNA and l/mHSP72 are higher among critically ill patients, further induced by HS, not by LPS. HSP72 proteins and HSP72 mRNA are related to serum ILs and are negatively related to supernatant cytokines, not being influenced by HSP72 polymorphisms, cortisol, or illness severity. Gln may depress l/mHSP72 after LPS exposure and enhance them after HS induction, but it may not affect early induced HSP72 mRNA.

  4. Hsp60 is targeted to a cryptic mitochondrion-derived organelle ("crypton") in the microaerophilic protozoan parasite Entamoeba histolytica. (United States)

    Mai, Z; Ghosh, S; Frisardi, M; Rosenthal, B; Rogers, R; Samuelson, J


    Entamoeba histolytica is a microaerophilic protozoan parasite in which neither mitochondria nor mitochondrion-derived organelles have been previously observed. Recently, a segment of an E. histolytica gene was identified that encoded a protein similar to the mitochondrial 60-kDa heat shock protein (Hsp60 or chaperonin 60), which refolds nuclear-encoded proteins after passage through organellar membranes. The possible function and localization of the amebic Hsp60 were explored here. Like Hsp60 of mitochondria, amebic Hsp60 RNA and protein were both strongly induced by incubating parasites at 42 degreesC. 5' and 3' rapid amplifications of cDNA ends were used to obtain the entire E. histolytica hsp60 coding region, which predicted a 536-amino-acid Hsp60. The E. histolytica hsp60 gene protected from heat shock Escherichia coli groEL mutants, demonstrating the chaperonin function of the amebic Hsp60. The E. histolytica Hsp60, which lacked characteristic carboxy-terminal Gly-Met repeats, had a 21-amino-acid amino-terminal, organelle-targeting presequence that was cleaved in vivo. This presequence was necessary to target Hsp60 to one (and occasionally two or three) short, cylindrical organelle(s). In contrast, amebic alcohol dehydrogenase 1 and ferredoxin, which are bacteria-like enzymes, were diffusely distributed throughout the cytosol. We suggest that the Hsp60-associated, mitochondrion-derived organelle identified here be named "crypton," as its structure was previously hidden and its function is still cryptic.

  5. Treatment of Arabidopsis thaliana seeds with an HSP90 inhibitor increases plant resistance (United States)

    Kozeko, Liudmyla


    Resistance of plants to unfavourable conditions is an important feature to use them as an autotrophic link of Life Support Systems in space exploration missions. It significantly depends on basic and stress-induced levels of heat shock proteins (HSP) in cells. It is known that HSP90 can bind and maintain heat shock transcription factors (HSF) as a monomer that lacks DNA binding activity and thereby regulate HSP expression. Modulation of activity of the HSP synthesis and resistance by HSP90 in plants is not well investigated. The objective of this study was to determine how treatment of seeds with an HSP90 inhibitor affects environmental responsiveness in Arabidopsis thaliana. Seed treatment with geldanamycin (GDA) was used to reduce HSP90 function. The affect of space flight stressors was simulated by gamma-irradiation and thermal upshift. Two series of experiments were carried out: 1) exposure of dry seeds to gamma-irradiation (1 kGy, ^{60}Co); 2) heat shock of seedlings. It was shown that GDA treatment of seeds stimulated the seedling growth after seed irradiation. It also increased both the basic thermotolerance (45°C for 45 min) and induced thermotolerance (45°C for 1,5-2,5 h after pretreatment at 37°C for 2 h) in seedlings. In addition, seed treatment with GDA had a prolonged effect on the HSP70 production in seedlings under normal and stressful conditions. It shows that the stimulatory effects of GDA may be caused by induction of HSP70 synthesis. The obtained data demonstrate that pre-treatment of seeds with GDA before planting allows inducing the stress resistance at least at early growth stages of plants.

  6. Polymorphism analysis of the hsp70 stress gene in Broiler chickens (Gallus gallus of different breeds

    Directory of Open Access Journals (Sweden)

    Carmen Maria Mazzi


    Full Text Available The promoter region and the beginning of the coding region of the hsp70 stress gene were analysed in broiler chickens of a commercial breed (Hubbard-Pettersen, a breed selected for weight gain (PP1 and a non-selected breed (naked-neck Label Rouge. The naked neck gene (Naked neck, Na, which reduces feathering in birds and is thus related to heat resistance, was present in both PP1 and Label Rouge breeds. Genomic DNA was restricted with PstI and Southern blotting analysis of the samples revealed the absence of polymorphic sites for that enzyme in the promoter region and beginning of the coding region of the hsp70 gene of studied birds. PCR-SSCP analysis of these regions, however, indicated the presence of polymorphisms in the beginning of the coding region and the sequencing of the PCR products confirmed and identified two polymorphic sites in this region: a transition A ® G in position +258 and a transversion C ® G in position +276. Both mutations were considered to be silent, since they did not modify the aminoacid sequence of the protein Hsp70. The promoter region of the hsp70 gene was identical in all studied birds, indicating that the regulation pattern of this gene must be the same in all birds at the promoter region. Three different alleles (hsp70-1, hsp70-2 and hsp70-3 were identified for the hsp70 gene from the observed mutations. The allele hsp70-3 was detected in only two breeds, Hubbard-Pettersen and PP1, but at a low frequency (0,016 and 0,006, respectively.

  7. Correlation of HSP110 expression with all-trans retinoic acid-induced apoptosis. (United States)

    Evrard, L; Vanmuylder, N; Dourov, N; Hermans, C; Biermans, J; Werry-Huet, A; Rooze, M; Louryan, S


    In a previous study, we observed the strong expression of a stress protein of the HSP100/Clp family (HSP110) in apoptotic mesectodermal cells during early mouse facial development. In the present study, we describe the strong expression of the same HSP110 in mesectodermal cells undergoing apoptosis after all-trans retinoic acid (RA) administration. We used a teratological model known to increase cell deaths mainly in the first and second branchial arches during mammalian cephalogenesis: the treatment of E9 mouse embryos with all-trans RA, which results in craniofacial malformations comparable to those that characterize mandibulofacial dysostosis in man. Pregnant NMRI mice were treated with 60 mg/kg body weight of all-trans RA, given orally on day 9 of gestation; embryos were taken 4, 12 or 24 hr after RA administration. The apoptotic pattern of RA-induced cell deaths was confirmed using the dUTP biotin nick-end labeling (TUNEL) method and transmission electron microscopy (TEM). HSP110 expression was detected using an immunohistochemical approach. The increase in the number of TUNEL-positive cells and HSP110-positive cells after all-trans RA administration was quantified in the first branchial arch using a computerized method. Twelve hours after RA administration, the increase in the number of HSP110-positive cells is greater than the increase in the number of TUNEL-positive cells. Twenty-four hours after RA administration, only TUNEL-positive cells remain strong in number. We suggest that HSP110 expression could represent a biochemical event of apoptotic cell death induced by RA, associated with early stages of the apoptotic process. In order to find out if HSP110 expression resulted from neosynthesis, we performed in situ hybridization, which demonstrated that the expression of HSP110 occurred at the level of mRNA.

  8. Serum HSP27 is associated with medullary perfusion in kidney allografts (United States)

    Marquez, Eva; Sadowski, Elizabeth; Reese, Shannon; Vidyasagar, Aparna; Artz, Nathan; Fain, Sean; Jacobson, Lynn; Swain, William; Djamali, Arjang


    Background Heat shock protein 27 (HSP27) is a small HSP up-regulated in response to stress in the kidney. The relationship between HSP27 and intrarenal oxygenation in patients with native and transplant kidney disease is unknown. Methods We compared HSP27 levels, intrarenal oxygenation measured by blood oxygen-level dependent (BOLD) imaging using R2* values, and perfusion determined by arterial spin labeling (ASL) magnetic resonance imaging (MRI), between patients with native and transplant kidney disease (n=28). Results There were no statistical differences in mean age (53.9 vs. 47.1 years), kidney function (63.6 vs. 50.7 ml/min per 1.73 m2), mean arterial blood pressure (91.6 vs. 91.1 mm Hg), hematocrit (40.6% vs. 39.3%), diuretic or angiotensin-converting enzyme inhibitor use, serum or urine levels of hydrogen peroxide, nitric oxide, F2 isoprostanes and HSP27 between native and transplant kidneys. BOLD-MRI studies demonstrated comparable patterns in intrarenal oxygen bioavailability (medullary R2* 18.1 vs. 18.3/s and cortical R2* 12 vs. 11.7/s, respectively). However, medullary perfusion was significantly lower in transplant kidneys (36.4 vs. 78.7 ml/100 g per minute, p=0.0002). There was a linear relationship between serum HSP27 concentrations and medullary perfusion in kidney allografts (HSP27 concentration [ng/mL] = 0.78 + 0.09 medullary perfusion, R2=0.43, p=0.01). Conclusions Our study demonstrates that medullary perfusion is significantly lower in kidney allografts compared with native kidneys with comparable renal function. We further noted a direct association between serum HSP27 levels and medullary perfusion after transplantation. Additional studies are needed to examine the role of HSP27 as a biomarker of kidney disease progression. PMID:22383348

  9. Probing molecular mechanisms of the Hsp90 chaperone: biophysical modeling identifies key regulators of functional dynamics.

    Directory of Open Access Journals (Sweden)

    Anshuman Dixit

    Full Text Available Deciphering functional mechanisms of the Hsp90 chaperone machinery is an important objective in cancer biology aiming to facilitate discovery of targeted anti-cancer therapies. Despite significant advances in understanding structure and function of molecular chaperones, organizing molecular principles that control the relationship between conformational diversity and functional mechanisms of the Hsp90 activity lack a sufficient quantitative characterization. We combined molecular dynamics simulations, principal component analysis, the energy landscape model and structure-functional analysis of Hsp90 regulatory interactions to systematically investigate functional dynamics of the molecular chaperone. This approach has identified a network of conserved regions common to the Hsp90 chaperones that could play a universal role in coordinating functional dynamics, principal collective motions and allosteric signaling of Hsp90. We have found that these functional motifs may be utilized by the molecular chaperone machinery to act collectively as central regulators of Hsp90 dynamics and activity, including the inter-domain communications, control of ATP hydrolysis, and protein client binding. These findings have provided support to a long-standing assertion that allosteric regulation and catalysis may have emerged via common evolutionary routes. The interaction networks regulating functional motions of Hsp90 may be determined by the inherent structural architecture of the molecular chaperone. At the same time, the thermodynamics-based "conformational selection" of functional states is likely to be activated based on the nature of the binding partner. This mechanistic model of Hsp90 dynamics and function is consistent with the notion that allosteric networks orchestrating cooperative protein motions can be formed by evolutionary conserved and sparsely connected residue clusters. Hence, allosteric signaling through a small network of distantly connected

  10. Down-regulation of HSP27 sensitizes TRAIL-resistant tumor cell to TRAIL-induced apoptosis

    DEFF Research Database (Denmark)

    Zhuang, Hongqin; Jiang, Weiwei; Cheng, Wei


    oxygen species or anticancer drugs. Their elevated expressions facilitate cells to survive in stress circumstances. The HSP27 expression is enhanced in many tumor cells, implying that it is involved in tumor progression and the development of treatment resistance in various tumors, including lung cancer...... siRNA on drug sensitization of A549 cells to TRAIL treatment. The results showed that treatment of A549 cells with HSP27 siRNA down-regulated HSP27 expression but did not induce significant apoptosis. However, combination of HSP27 siRNA with TRAIL-induced significant apoptosis in TRAIL-resistant A549...... cells. In addition to inducing caspases activation and apoptosis, combined treatment with HSP27 siRNA and TRAIL also increased JNK and p53 expression and activity. Collectively, these findings provide a conclusion that siRNA targeting of the HSP27 gene specifically down-regulated HSP27 expression in A...

  11. Bio-inspired vision (United States)

    Posch, C.


    Nature still outperforms the most powerful computers in routine functions involving perception, sensing and actuation like vision, audition, and motion control, and is, most strikingly, orders of magnitude more energy-efficient than its artificial competitors. The reasons for the superior performance of biological systems are subject to diverse investigations, but it is clear that the form of hardware and the style of computation in nervous systems are fundamentally different from what is used in artificial synchronous information processing systems. Very generally speaking, biological neural systems rely on a large number of relatively simple, slow and unreliable processing elements and obtain performance and robustness from a massively parallel principle of operation and a high level of redundancy where the failure of single elements usually does not induce any observable system performance degradation. In the late 1980`s, Carver Mead demonstrated that silicon VLSI technology can be employed in implementing ``neuromorphic'' circuits that mimic neural functions and fabricating building blocks that work like their biological role models. Neuromorphic systems, as the biological systems they model, are adaptive, fault-tolerant and scalable, and process information using energy-efficient, asynchronous, event-driven methods. In this paper, some basics of neuromorphic electronic engineering and its impact on recent developments in optical sensing and artificial vision are presented. It is demonstrated that bio-inspired vision systems have the potential to outperform conventional, frame-based vision acquisition and processing systems in many application fields and to establish new benchmarks in terms of redundancy suppression/data compression, dynamic range, temporal resolution and power efficiency to realize advanced functionality like 3D vision, object tracking, motor control, visual feedback loops, etc. in real-time. It is argued that future artificial vision systems

  12. Bio-oil fractionation and condensation

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Robert C.; Jones, Samuel T.; Pollard, Anthony


    The present invention relates to a method of fractionating bio-oil vapors which involves providing bio-oil vapors comprising bio-oil constituents. The bio-oil vapors are cooled in a first stage which comprises a condenser having passages for the bio-oil separated by a heat conducting wall from passages for a coolant. The coolant in the condenser of the first stage is maintained at a substantially constant temperature, set at a temperature in the range of 75 to C., to condense a first liquid fraction of liquefied bio-oil constituents in the condenser of the first stage. The first liquid fraction of liquified bio-oil constituents from the condenser in the first stage is collected. Also disclosed are steps for subsequently recovering further liquid fractions of liquefied bio-oil constituents. Particular compositions of bio-oil condensation products are also described.

  13. Bio-oil fractionation and condensation (United States)

    Brown, Robert C; Jones, Samuel T; Pollard, Anthony


    A method of fractionating bio-oil vapors which involves providing bio-oil vapors comprising bio-oil constituents is described. The bio-oil vapors are cooled in a first stage which comprises a condenser having passages for the bio-oil separated by a heat conducting wall from passages for a coolant. The coolant in the condenser of the first stage is maintained at a substantially constant temperature, set at a temperature in the range of 75 to C., to condense a first liquid fraction of liquefied bio-oil constituents in the condenser of the first stage. The first liquid fraction of liquified bio-oil constituents from the condenser in the first stage is collected. Also described are steps for subsequently recovering further liquid fractions of liquefied bio-oil constituents. Particular compositions of bio-oil condensation products are also described.

  14. HSP70 expression in the copper butterfly Lycaena tityrus across altitudes and temperatures

    DEFF Research Database (Denmark)

    Karl, I.; Sørensen, Jesper Givskov; Loeschcke, Volker


    The ability to express heat-shock proteins (HSP) under thermal stress is an essential mechanism for ectotherms to cope with unfavourable conditions. In this study, we investigate if Copper butterflies originating from different altitudes and/or being exposed to different rearing and induction...... temperatures show differences in HSP70 expression. HSP70 expression increased substantially at the higher rearing temperature in low-altitude butterflies, which might represent an adaptation to occasionally occurring heat spells. On the other hand, high-altitude butterflies showed much less plasticity...

  15. Hsp27gene in Drosophila ananassae subgroup was split by a recently acquired intron

    Indian Academy of Sciences (India)



    InDrosophila , heat shock protein 27 (Hsp27) is a critical single-copy intron-free nuclear gene involved in the defense responseagainst fungi and bacteria, and is a regulator of adult lifespan. In the present study, 33 homologousHsp27nucleotide sequencesfrom differentDrosophilaspecies were amplified by PCR and reverse transcription PCR, and the phylogenetic relationshipswere analysed using neighbour-joining, maximum-likelihood and Bayesian methods. The phylogenetic topologies from anal-ysis with different algorithms were similar, suggesting that theHsp27gene was split by a recently acquired intron during theevolution of theDrosophila ananassaesubgroup

  16. Reactive oxygen species generated by a heat shock protein (Hsp) inducing product contributes to Hsp70 production and Hsp70-mediated protective immunity in Artemia franciscana against pathogenic vibrios. (United States)

    Baruah, Kartik; Norouzitallab, Parisa; Linayati, Linayati; Sorgeloos, Patrick; Bossier, Peter


    The cytoprotective role of heat shock protein (Hsp70) described in a variety of animal disease models, including vibriosis in farmed aquatic animals, suggests that new protective strategies relying upon the use of compounds that selectively turn on Hsp genes could be developed. The product Tex-OE® (hereafter referred to as Hspi), an extract from the skin of the prickly pear fruit, Opuntia ficus indica, was previously shown to trigger Hsp70 synthesis in a non-stressful situation in a variety of animals, including in a gnotobiotically (germ-free) cultured brine shrimp Artemia franciscana model system. This model system offers great potential for carrying out high-throughput, live-animal screens of compounds that have health benefit effects. By using this model system, we aimed to disclose the underlying cause behind the induction of Hsp70 by Hspi in the shrimp host, and to determine whether the product affects the shrimp in inducing resistance towards pathogenic vibrios. We provide unequivocal evidences indicating that during the pretreatment period with Hspi, there is an initial release of reactive oxygen species (hydrogen peroxide and/or superoxide anion), generated by the added product, in the rearing water and associated with the host. The reactive molecules generated are the triggering factors responsible for causing Hsp70 induction within Artemia. We have also shown that Hspi acts prophylactically at an optimum dose regimen to confer protection against pathogenic vibrios. This salutary effect was associated with upregulation of two important immune genes, prophenoloxidase and transglutaminase of the innate immune system. These findings suggest that inducers of stress protein (e.g. Hsp70) are potentially important modulator of immune responses and might be exploited to confer protection to cultured shrimp against Vibrio infection.

  17. Subcellular fractionation reveals HSP72 does not associate with SERCA in human skeletal muscle following damaging eccentric and concentric exercise. (United States)

    Frankenberg, Noni T; Lamb, Graham D; Vissing, Kristian; Murphy, Robyn M


    Through its upregulation and/or translocation, heat shock protein 72 (HSP72) is involved in protection and repair of key proteins after physiological stress. In human skeletal muscle we investigated HSP72 protein after eccentric (ECC1) and concentric (CONC) exercise and repeated eccentric exercise (ECC2; 8 wk later) and whether it translocated from its normal cytosolic location to membranes/myofibrils. HSP72 protein increased ~2-fold 24 h after ECC1, with no apparent change after CONC or ECC2. In resting (nonstressed) human skeletal muscle the total pool of HSP72 protein was present almost exclusively in the cytosolic fraction, and after each exercise protocol the distribution of HSP72 protein remained unaltered. Overall, the amount of HSP72 protein in the cytosol increased 24 h after ECC1, matching the fold increase that was measured in total HSP72 protein. To better ascertain the capabilities and limitations of HSP72, using quantitative Western blotting we determined the HSP72 protein content to be 11.4 μmol/kg wet weight in resting human vastus lateralis muscle, which is comprised of Type I (slow-twitch) and Type II (fast-twitch) fibers. HSP72 protein content was similar in individual Type I or II fiber segments. After physiological stress, HSP72 content can increase and, although the functional consequences of increased amounts of HSP72 protein are poorly understood, it has been shown to bind to and protect protein pumps like SERCA and Na(+)-K(+)-ATPase. Given no translocation of cytosolic HSP72, these findings suggest eccentric contractions, unlike other forms of stress such as heat, do not trigger tight binding of HSP72 to its primary membrane-bound target proteins, in particular SERCA.

  18. Expression and Significance of HSP70 and MGMT in Breast Neoplasms%HSP70和MGMT在乳腺肿瘤中的表达和意义

    Institute of Scientific and Technical Information of China (English)

    吴荣薇; 薄爱华; 王发亮; 芦广萍


    目的:探讨乳腺肿瘤组织中热休克蛋白HSP70和DNA修复酶O6-甲基鸟嘌呤-DNA甲基转移酶(O6-methyguanine-DNA methytransferase,MGMT)的表达及临床意义.方法:采用免疫组织化学SABC法检测65例乳腺肿瘤组织中HSP70和MGMT的表达情况.结果:乳腺肿瘤中HSP70的阳性率为80.00%(52/65),MGMT的阳性率为60.00%(39/65),二者呈中度相关性(r=0.46).其中,有淋巴结转移组的阳性率高于无转移组(P<0.05).结论:HSP70和MGMT的表达率与淋巴结转移情况有关.检测HSP70与MGMT对乳腺肿瘤预后和化疗方案的选择有指导意义.

  19. Ionizing radiation improves glioma-specific targeting of superparamagnetic iron oxide nanoparticles conjugated with cmHsp70.1 monoclonal antibodies (SPION-cmHsp70.1) (United States)

    Shevtsov, Maxim A.; Nikolaev, Boris P.; Ryzhov, Vyacheslav A.; Yakovleva, Ludmila Y.; Marchenko, Yaroslav Y.; Parr, Marina A.; Rolich, Valerij I.; Mikhrina, Anastasiya L.; Dobrodumov, Anatolii V.; Pitkin, Emil; Multhoff, Gabriele


    The stress-inducible 72 kDa heat shock protein Hsp70 is known to be expressed on the membrane of highly aggressive tumor cells including high-grade gliomas, but not on the corresponding normal cells. Membrane Hsp70 (mHsp70) is rapidly internalized into tumor cells and thus targeting of mHsp70 might provide a promising strategy for theranostics. Superparamagnetic iron oxide nanoparticles (SPIONs) are contrast negative agents that are used for the detection of tumors with MRI. Herein, we conjugated the Hsp70-specific antibody (cmHsp70.1) which is known to recognize mHsp70 to superparamagnetic iron nanoparticles to assess tumor-specific targeting before and after ionizing irradiation. In vitro experiments demonstrated the selectivity of SPION-cmHsp70.1 conjugates to free and mHsp70 in different tumor cell types (C6 glioblastoma, K562 leukemia, HeLa cervix carcinoma) in a dose-dependent manner. High-resolution MRI (11 T) on T2-weighted images showed the retention of the conjugates in the C6 glioma model. Accumulation of SPION-cmHsp70.1 nanoparticles in the glioma resulted in a nearly 2-fold drop of values in comparison to non-conjugated SPIONs. Biodistribution analysis using NLR-M2 measurements showed a 7-fold increase in the tumor-to-background (normal brain) uptake ratio of SPION-cmHsp70.1 conjugates in glioma-bearing rats in comparison to SPIONs. This accumulation within Hsp70-positive glioma was further enhanced after a single dose (10 Gy) of ionizing radiation. Elevated accumulation of the magnetic conjugates in the tumor due to radiosensitization proves the combination of radiotherapy and application of Hsp70-targeted agents in brain tumors.The stress-inducible 72 kDa heat shock protein Hsp70 is known to be expressed on the membrane of highly aggressive tumor cells including high-grade gliomas, but not on the corresponding normal cells. Membrane Hsp70 (mHsp70) is rapidly internalized into tumor cells and thus targeting of mHsp70 might provide a promising strategy

  20. Co-effects of extremely low frequency electromagnetic field (ELF-EMF) and temper-ature on hsp22 and hsp26 expression in Drosophila melanogaster%高温下极低频电磁场暴露对果蝇hsp22和hsp26基因影响的研究

    Institute of Scientific and Technical Information of China (English)

    张晶; 张紫燕; 杨传俊; 蔡澎


    Objective To study the combined effect of extremely low frequency electromagnetic fields ( ELF-EMF ) exposure (50 Hz and 3 mT) and high temperature exposure on hsp22 and hsp26 expression in Drosophila melanogaster. Methods Under the conditions of 50 Hz and 3 mT ELF-EMF,the male and female D.melanogaster was separately exposed for 2 h at 25℃, 27.5℃, 30℃, 32.5℃,and 35℃, respectively.The death rate, exercise capacity and the hsp22 and hsp26 expression levels of D.melanogaster were detected after exposure;or at 25℃and 35℃at 2 h, 4 h, 8 h, and 24 h, respectively.The hsp22 and hsp26 expression levels of D.melanogaster were tested after exposure .Results After exposure for 2 h at 25℃, 27.5℃, 30℃, 32.5℃, and 35℃, temperature had a significant effect (P0.05).After exposure for 2 h at 25℃, 27.5℃, 30℃, 32.5℃, and 35℃, the hsp22 and hsp26 expression levels of D.melanogaster were significantly impacted(P<0.01) by gender, temperature, ELF-EMF and exposure time, and tem-perature was the most important factor .Gender, temperature, ELF-EMF and exposure time had some interaction with the results.Conclusion ELF-EMF can influence the expression levels of hsp22 and hsp26.ELF-EMF exposure contributes to heat tolerance in D.melanogaster through accelerated expression of hsp22 and hsp26.%目的:探讨在高温(35℃)条件下,极低频电磁场( extremely low frequency electromagnetic fields ,ELF-EMF)对雌雄黑腹果蝇hsp22和hsp26基因表达水平的影响。方法在25、27.5、30、32.5、35℃下,1日龄雌、雄黑腹果蝇在50 Hz、3 mT ELF-EMF暴露2 h,测定其死亡率、运动能力和hsp22、hsp26表达水平;在25和35℃下,1日龄雌、雄黑腹果蝇在50 Hz、3 mT ELF-EMF暴露2、4、8、24 h,测定其hsp22、hsp26表达水平。结果在25、27.5、30、32.5、35℃下暴露2 h,温度变化对果蝇的死亡率、运动能力和hsp22、hsp26表达水平有极显著影响(P<0.01),ELF-EMF对hsp

  1. Bio-nanopatterning of Surfaces

    Directory of Open Access Journals (Sweden)

    Yeung Chun


    Full Text Available AbstractBio-nanopatterning of surfaces is a very active interdisciplinary field of research at the interface between biotechnology and nanotechnology. Precise patterning of biomolecules on surfaces with nanometre resolution has great potential in many medical and biological applications ranging from molecular diagnostics to advanced platforms for fundamental studies of molecular and cell biology. Bio-nanopatterning technology has advanced at a rapid pace in the last few years with a variety of patterning methodologies being developed for immobilising biomolecules such as DNA, peptides, proteins and viruses at the nanoscale on a broad range of substrates. In this review, the status of research and development are described, with particular focus on the recent advances on the use of nanolithographic techniques as tools for biomolecule immobilisation at the nanoscale. Present strengths and weaknesses, as well future challenges on the different nanolithographic bio-nanopatterning approaches are discussed.

  2. Evaluation of Emissions Bio diesel

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez Maroto, J. J.; Dorronsoro Arenal, J. L.; Rojas Garcia, E.; Perez Pastor, R.; Garcia Alonso, S.


    The generation of energy from vegetal products is one of the possibilities to our reach in order to reduce the atmospheric pollution. Particularly, the use of bio diesel in internal combustion engines can be one of the best options. The finest particles emitted by the combustion engines are easily breathable and on them different substances can be absorbed presumably toxic, between which it is possible to emphasize the polycyclic aromatic hydrocarbons (PAHs), by its demonstrated carcinogen character. In this work, it is studied on the one hand, the characteristics that can present the aerosol of emission in a diesel engine with a maximum power of 97 kW, working without load to 600 rpm, using as combustible mixtures of bio diesel and diesel in different proportions. On the other hand, the evolution that takes place in the concentration of PAHs in emission particles, according to the percentage of bio diesel used in the combustible mixture. (Author) 9 refs.

  3. Small heat shock protein Hsp27 is required for proper heart tube formation. (United States)

    Brown, Daniel D; Christine, Kathleen S; Showell, Christopher; Conlon, Frank L


    The small heat shock protein Hsp27 has been shown to be involved in a diverse array of cellular processes, including cellular stress response, protein chaperone activity, regulation of cellular glutathione levels, apoptotic signaling, and regulation of actin polymerization and stability. Furthermore, mutation within Hsp27 has been associated with the human congenital neuropathy Charcot-Marie Tooth (CMT) disease. Hsp27 is known to be expressed in developing embryonic tissues; however, little has been done to determine the endogenous requirement for Hsp27 in developing embryos. In this study, we show that depletion of XHSP27 protein results in a failure of cardiac progenitor fusion resulting in cardia bifida. Furthermore, we demonstrate a concomitant disorganization of actin filament organization and defects in myofibril assembly. Moreover, these defects are not associated with alterations in specification or differentiation. We have thus demonstrated a critical requirement for XHSP27 in developing cardiac and skeletal muscle tissues.

  4. Trichostatin A Extends the Lifespan of Drosophila melanogaster by Elevating hsp22 Expression

    Institute of Scientific and Technical Information of China (English)

    Dan TAO; Jun LU; Hui SUN; Yan-Mei ZHAO; Zhi-Gen YUAN; Xiao-Xue LI; Bai-Qu HUANG


    The level of acetylation of histones in nucleosomes is related to the longevity of yeast and animals. However, the mechanisms by which acetylation and deacetylation affect longevity remain unclear.In present study, we investigated the influence of histone acetylation modification on the expression of hsp22gene and the lifespan in Drosophila melanogaster using histone deacetylase (HDAC) inhibitor Trichostatin A (TSA). The results showed that TSA could extend the lifespan of Drosophila melanogaster. Furthermore,TSA significantly promoted the hsp22 gene transcription, and affected the chromatin morphology at the locus of hsp22 gene along the polytene chromosome. Present data implicate that TSA may affect the lifespan of Drosophila through changing the level of histone acetylation and influencing the expression of hsp22 gene that is related to aging.

  5. A review of recent patents on the protozoan parasite HSP90 as a drug target. (United States)

    Angel, Sergio O; Matrajt, Mariana; Echeverria, Pablo C


    Diseases caused by protozoan parasites are still an important health problem. These parasites can cause a wide spectrum of diseases, some of which are severe and have high morbidity or mortality if untreated. Since they are still uncontrolled, it is important to find novel drug targets and develop new therapies to decrease their remarkable social and economic impact on human societies. In the past years, human HSP90 has become an interesting drug target that has led to a large number of investigations both at state organizations and pharmaceutical companies, followed by clinical trials. The finding that HSP90 has important biological roles in some protozoan parasites like Plasmodium spp, Toxoplasma gondii and trypanosomatids has allowed the expansion of the results obtained in human cancer to these infections. This review summarizes the latest important findings showing protozoan HSP90 as a drug target and presents three patents targeting T. gondii, P. falciparum and trypanosomatids HSP90.

  6. Changing of the HSP70 Content in the Baikal Endemic Sponges Lubomirskiidae Under Conditions of Hyperthermia

    Directory of Open Access Journals (Sweden)

    Itskovich V.B.


    Full Text Available Baikal endemic sponges (Lubomirskiidae make up the bulk of the benthos biomass of the lake. For the first time the changes in the content of HSP70 in response to elevated environment temperature were analyzed in three endemic species of Baikal sponges: Baikalospongia bacillifera (Dybowski, 1880, B. intermedia (Dybowski, 1880 and Swartschewskia papyracea (Dybowski, 1880. Interspecific variability of constitutive HSP70 level was revealed for representatives of the three analyzed Lubomirskiidae species. After exposure at 13 °С for 3 and 7 days opposite changes were noted in the amount of HSP70. Under conditions of hyperthermia the protein level decrease at Baikalospongia species, while at the S. papyracea HSP70 content slightly increased. The differences in the mechanisms of stress adaptation probably affect the thermal resistance of the species, as well as are evidence supporting their specific status.

  7. Immunohistochemical changes in the expression of HSP27 in exercised human vastus lateralis muscle

    DEFF Research Database (Denmark)

    Folkesson, M; Mackey, Abigail; Holm, L;


    . METHODS: Two different exercise protocols were used: (1) one-leg ergometer cycling (EC, n = 6) consisting of two 30-min bouts at 40% and 75% of peak oxygen uptake, respectively, and (2) leg extension resistance exercise (RE, n = 9) including 10 sets of eight repetitions at a load corresponding to 70......AIM: The role of HSP27 in the adaptive process of skeletal muscle to exercise, especially in humans, is not well understood. The objective of this study was to investigate immunohistochemical changes in HSP27 expression in human vastus lateralis muscle following resistance and endurance exercises......% of one maximal repetition (1RM). Immunohistochemistry using specific monoclonal antibodies was used to determine the location of HSP27 protein in muscle biopsies from human vastus lateralis. RESULTS: Our results show that RE, but not EC, induced a significant appearance of scattered accumulations of HSP...

  8. Hsp60 and p70S6K form a complex in human cardiomyocytes

    Directory of Open Access Journals (Sweden)

    Kroupskaya I. V.


    Full Text Available Molecular chaperon Hsp60 and protein kinase p70S6K play an important functional role in the regulation of cardiomyocytes vital function or apoptosis. Aim. To study a possibility of in vivo complex formation between Hsp60 and p70S6K in cardiomyocytes. Methods. Co-immunoprecipitation, Western-blot analysis. Results. We have identified in vivo interaction between molecular chaperone Hsp60 and two isoforms of proteinkinase p70S6K in human myocardium, normal and affected by cardiomyopathy. Conclusions. The results obtained suggest a possible participation of molecular chaperon Hsp60 in regulation of p70S6K activity in stressinduced apoptotic signaling pathway in cardiomyocytes.

  9. Molecular characterization, gene expression and transcriptional regulation of cytosolic HSP90 genes in the flatfish Senegalese sole (Solea senegalensis Kaup). (United States)

    Manchado, Manuel; Salas-Leiton, Emilio; Infante, Carlos; Ponce, Marian; Asensio, Esther; Crespo, Aniela; Zuasti, Eugenia; Cañavate, José Pedro


    HSP90 proteins are chaperones that play a pivotal role in controlling multiple regulatory pathways such as stress defense, hormone signalling, cell cycle control, cell proliferation and differentiation, and apoptosis. In this study, two cDNAs encoding for cytosolic HSP90, referred to as HSP90AA and HSP90AB, have been sequenced. Main features and sequence identities with other fish and mammals are described. Phylogenetic analysis grouped both genes into two separate clusters with their fish and mammalian counterparts. Expression profiles during larval development and in juvenile tissues were analyzed using a real-time PCR approach. In juvenile fish, HSP90AB was constitutively expressed with lower transcript levels in skeletal muscle. In contrast, HSP90AA was mainly expressed in heart, skeletal muscle and skin. During metamorphosis, HSP90AB mRNA levels did not change whereas HSP90AA transcripts decreased significantly at the beginning of metamorphosis with the lowest mRNA levels at the metamorphosis climax. Due to the role of thyroid hormones (THs) on sole metamorphosis, the transcriptional regulation of HSP90 genes by THs was evaluated. Larvae exposed to the goitrogen thiourea (TU) exhibited higher HSP90AA mRNA levels than untreated control. Moreover, adding exogenous T4 hormone to TU-treated larvae restored the steady-state levels with respect to the untreated control. Unlike HSP90AA, the transcript levels of HSP90AB did not vary under any treatments. The response of both HSP90 genes to thermal stress in post-metamorphic individuals was also studied. A heat shock treatment (+7.9 degrees C for 1 h) rapidly activated HSP90AA (but not HSP90AB) transcription, reaching a peak after 30 min and declining expression levels progressively in the following 24 h. No significant changes in HSP90AA or HSP90AB transcript levels after a cold shock (-10 degrees C for 1 h) were observed. Overall, these results demonstrate that HSP90AA transcription is down-regulated by THs and up

  10. A rat retinal damage model predicts for potential clinical visual disturbances induced by Hsp90 inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Dan, E-mail: [Synta Pharmaceuticals Corp., 45 Hartwell Avenue, Lexington, MA 02421 (United States); Liu, Yuan; Ye, Josephine; Ying, Weiwen; Ogawa, Luisa Shin; Inoue, Takayo; Tatsuta, Noriaki; Wada, Yumiko; Koya, Keizo [Synta Pharmaceuticals Corp., 45 Hartwell Avenue, Lexington, MA 02421 (United States); Huang, Qin [Department of Pathology and Laboratory Medicine, Veterans Affairs Boston Healthcare System, 1400 VFW Parkway, West Roxbury, MA 02132 (United States); Bates, Richard C.; Sonderfan, Andrew J. [Synta Pharmaceuticals Corp., 45 Hartwell Avenue, Lexington, MA 02421 (United States)


    In human trials certain heat shock protein 90 (Hsp90) inhibitors, including 17-DMAG and NVP-AUY922, have caused visual disorders indicative of retinal dysfunction; others such as 17-AAG and ganetespib have not. To understand these safety profile differences we evaluated histopathological changes and exposure profiles of four Hsp90 inhibitors, with or without clinical reports of adverse ocular effects, using a rat retinal model. Retinal morphology, Hsp70 expression (a surrogate marker of Hsp90 inhibition), apoptotic induction and pharmacokinetic drug exposure analysis were examined in rats treated with the ansamycins 17-DMAG and 17-AAG, or with the second-generation compounds NVP-AUY922 and ganetespib. Both 17-DMAG and NVP-AUY922 induced strong yet restricted retinal Hsp70 up-regulation and promoted marked photoreceptor cell death 24 h after the final dose. In contrast, neither 17-AAG nor ganetespib elicited photoreceptor injury. When the relationship between drug distribution and photoreceptor degeneration was examined, 17-DMAG and NVP-AUY922 showed substantial retinal accumulation, with high retina/plasma (R/P) ratios and slow elimination rates, such that 51% of 17-DMAG and 65% of NVP-AUY922 present at 30 min post-injection were retained in the retina 6 h post-dose. For 17-AAG and ganetespib, retinal elimination was rapid (90% and 70% of drugs eliminated from the retina at 6 h, respectively) which correlated with lower R/P ratios. These findings indicate that prolonged inhibition of Hsp90 activity in the eye results in photoreceptor cell death. Moreover, the results suggest that the retina/plasma exposure ratio and retinal elimination rate profiles of Hsp90 inhibitors, irrespective of their chemical class, may predict for ocular toxicity potential. - Highlights: • In human trials some Hsp90 inhibitors cause visual disorders, others do not. • Prolonged inhibition of Hsp90 in the rat eye results in photoreceptor cell death. • Retina/plasma ratio and retinal

  11. Sensitization of multidrug-resistant human cancer cells to Hsp90 inhibitors by down-regulation of SIRT1. (United States)

    Kim, Hak-Bong; Lee, Su-Hoon; Um, Jee-Hyun; Oh, Won Keun; Kim, Dong-Wan; Kang, Chi-Dug; Kim, Sun-Hee


    The effectiveness of Hsp90 inhibitors as anticancer agents was limited in multidrug-resistant (MDR) human cancer cells due to induction of heat shock proteins (Hsps) such as Hsp70/Hsp27 and P-glycoprotein (P-gp)-mediated efflux. In the present study, we showed that resistance to Hsp90 inhibitors of MDR human cancer cells could be overcome with SIRT1 inhibition. SIRT1 knock-down or SIRT1 inhibitors (amurensin G and EX527) effectively suppressed the resistance to Hsp90 inhibitors (17-AAG and AUY922) in several MDR variants of human lymphoblastic leukemia and human breast cancer cell lines. SIRT1 inhibition down-regulated the expression of heat shock factor 1 (HSF1) and subsequently Hsps and facilitated Hsp90 multichaperone complex disruption via hyperacetylation of Hsp90/Hsp70. These findings were followed by acceleration of ubiquitin ligase CHIP-mediated mutant p53 (mut p53) degradation and subsequent down-regulation of P-gp in 17-AAG-treated MDR cancer cells expressing P-gp and mut p53 after inhibition of SIRT1. Therefore, combined treatment with Hsp90 inhibitor and SIRT1 inhibitor could be a more effective therapeutic approach for Hsp90 inhibitor-resistant MDR cells via down-regulation of HSF1/Hsps, mut p53 and P-gp.

  12. Tat-HSP22 inhibits oxidative stress-induced hippocampal neuronal cell death by regulation of the mitochondrial pathway. (United States)

    Jo, Hyo Sang; Kim, Dae Won; Shin, Min Jea; Cho, Su Bin; Park, Jung Hwan; Lee, Chi Hern; Yeo, Eun Ji; Choi, Yeon Joo; Yeo, Hyeon Ji; Sohn, Eun Jeong; Son, Ora; Cho, Sung-Woo; Kim, Duk-Soo; Yu, Yeon Hee; Lee, Keun Wook; Park, Jinseu; Eum, Won Sik; Choi, Soo Young


    Oxidative stress plays an important role in the progression of various neuronal diseases including ischemia. Heat shock protein 22 (HSP22) is known to protect cells against oxidative stress. However, the protective effects and mechanisms of HSP22 in hippocampal neuronal cells under oxidative stress remain unknown. In this study, we determined whether HSP22 protects against hydrogen peroxide (H2O2)-induced oxidative stress in HT-22 using Tat-HSP22 fusion protein. We found that Tat-HSP22 transduced into HT-22 cells and that H2O2-induced cell death, oxidative stress, and DNA damage were significantly reduced by Tat-HSP22. In addition, Tat-HSP22 markedly inhibited H2O2-induced mitochondrial membrane potential, cytochrome c release, cleaved caspase-3, and Bax expression levels, while Bcl-2 expression levels were increased in HT-22 cells. Further, we showed that Tat-HSP22 transduced into animal brain and inhibited cleaved-caspase-3 expression levels as well as significantly inhibited hippocampal neuronal cell death in the CA1 region of animals in the ischemic animal model. In the present study, we demonstrated that transduced Tat-HSP22 attenuates oxidative stress-induced hippocampal neuronal cell death through the mitochondrial signaling pathway and plays a crucial role in inhibiting neuronal cell death, suggesting that Tat-HSP22 protein may be used to prevent oxidative stress-related brain diseases including ischemia.

  13. Determination of kinetic parameters in Tl dosemeters of LiF: Mg, Cu, P + PTFE developed in the ININ; Determinacion de parametros cineticos en dosimetros Tl de LiF: Mg, Cu, P + PTFE desarrollados en el ININ

    Energy Technology Data Exchange (ETDEWEB)

    Basurto G, B.S


    The objective of this work, is the one of determining the kinetic parameters of the dosemeter of LiF: Mg, Cu, P + Ptfe; starting from the curves Tl obtained at being irradiated with alpha radiation ({alpha}), beta ({beta}) and gamma ({gamma}). As like to compare its sensitivity with each radiation type, considering the sensitivity of the TLD-100 as the unit. In the Chapter 1, the fundamental structure of the matter is described, making emphasis in the different radiation types, and their interaction with this. In the Chapter 2, the units are described but used in the dosimetry of the radiation. In the Chapter 3, the basic concepts of the phenomenon of Tl are described and those are explained characteristic of the deconvolution method to determine the kinetics of the one phenomenon. In the Chapter 4, the methodology is detailed that was used in the elaboration of this thesis work, describing the material Tl that were considered like reference, as well as the sources of ionizing radiation, with those that the dosemeters were irradiated and the equipment in the one that the curves Tl was obtained. Reference is made to the software used to carry out the deconvolution of the curves Tl that were obtained in the one experimental development. In the Chapter 5, the obtained results of this study are presented, showing the tables of homogenization of dosemeters and the reading of the same one; they are observed the curves Tl obtained to different radiation doses (alpha, beta and gamma), the intensity Tl in function of the dose. Also they are tabulated, the obtained results in the kinetic parameters of the three different study materials (TLD-100H, USA; TLD-100, USA and LiF: Mg, Cu, P + Ptfe developed in the l.N.l.N). They are analyzed shortly for each material Tl their sensitivity to the ionizing radiation as well as their kinetic parameters. The obtained results showed that the Tl dosemeters of LiF: Mg,Cu,P + Ptfe, they presented a bigger sensitivity that the TLD-100 when

  14. Pulsed X-rays for interventional radiology: tests on active personal dosemeters (APD) (European project FP7 ORAMED WP3); Rayons X pulses pour la radiologie interventionnelle: tests sur dosimetres personnel actifs

    Energy Technology Data Exchange (ETDEWEB)

    Denoziere, M.; Bordy, J.M.; Daures, J.; Lecerf, N


    this report presents the results of the tests performed on Active Personal dosemeters (A.P.D.) to check their responses in pulsed X-ray beam used in interventional X-ray radiology. this work is one of the goal of O.R.A.M.E.D W.P.3. (Optimization of radiation protection for medical staff)The response of seven A.P.D.s types was measured in terms of dose equivalent Hp (10) for different pulsed X-ray width and dose rate. (author)

  15. Obtention of thermoluminescent efficiencies by means of irradiation of TLD-100 dosemeters with proton beams helium and carbon; Obtencion de eficiencias termoluminiscentes mediante irradiacion de dosimetros TLD-100 con haces de protones, helios y carbon

    Energy Technology Data Exchange (ETDEWEB)

    Avila, O.; Rodriguez V, M.; Aviles, P.; Gamboa de Buen, I.; Buenfil, A.E.; Ruiz T, C.; Brandan, M.E. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)


    In this work, the advances of a serial of measurements of relative efficiency thermoluminescent of heavy charged particles (PCP) with respect to gamma radiation for TLD-100, dosemeters of LiF: Mg,Ti manufactured by the Harshaw-Bicron company are reported. The PCP are essentials in the implementation of dosimetry associated with medical applications. The measurements before gamma radiation were carrying out using the Vickrad irradiator of the National Institute of Nuclear Research at dose of 1.663 Gy. The measures which are reported about protons, helium and carbon were realized using the Pelletron accelerator of the Physics Institute of the UNAM. (Author)

  16. The HSP90 inhibitor alvespimycin enhances the potency of telomerase inhibition by imetelstat in human osteosarcoma


    Hu, Yafang; Bobb, Daniel; He, Jianping; Hill, D. Ashley; Dome, Jeffrey S.


    The unsatisfactory outcomes for osteosarcoma necessitate novel therapeutic strategies. This study evaluated the effect of the telomerase inhibitor imetelstat in pre-clinical models of human osteosarcoma. Because the chaperone molecule HSP90 facilitates the assembly of telomerase protein, the ability of the HSP90 inhibitor alvespimycin to potentiate the effect of the telomerase inhibitor was assessed. The effect of single or combined treatment with imetelstat and alvespimycin on long-term grow...

  17. Heat Shock Protein 90 (HSP90 and Her2 in Adenocarcinomas of the Esophagus

    Directory of Open Access Journals (Sweden)

    Julia Slotta-Huspenina


    Full Text Available Her2 overexpression and amplification can be found in a significant subset of esophageal adenocarcinomas. The activity of Her2 has been shown to be modulated by molecular chaperones such as HSP90. We analyzed expression/amplification data for HSP90 and Her2 on 127 primary resected esophageal adenocarcinomas in order to evaluate a possible relationship between these two molecules. HSP90 expression determined by immunohistochemistry was observed in various levels. Thirty nine (39 tumors (30.7% were classified as Her2-positive according to their immunoreactivity and amplification status. There was a significant correlation between HSP90 expression and Her2-status (p = 0.008. This could also be demonstrated by quantitative protein expression analysis with reverse phase protein arrays (r = 0.9; p < 0.001. Her2-status was associated withpT-category (p = 0.041, lymph node metastases (p = 0.049 and tumor differentiation (p = 0.036 with a higher percentage of cases with negative Her2 status in lower tumor stagesA negative Her2-status was also associated with better survival in univariate and multivariate analysis (p = 0.001 and p = 0.014. For HSP90, no associations between clinical and pathological parameters were found. The observed association between HSP90 expression and Her2 suggests a co-regulation of these molecules in at least a subset of esophageal adenocarcinomas. Anti-HSP90 drugs, which recently have been introduced in cancer treatment, may also be an option for these tumors by targeting HSP90 alone or in combination with Her2.

  18. Detection of Leishmania spp. based on the gene encoding HSP20


    Montalvo, Ana M; Departamento de Parasitología, Instituto de Medicina Tropical Pedro Kourí. La Habana, Cuba.; Fraga, Jorge; Departamento de Parasitología, Instituto de Medicina Tropical Pedro Kourí. La Habana, Cuba.; Rodríguez, Omaira; Laboratorio de referencia e investigación en enfermedades tropicales de sanidad militar. Bogotá, Colombia.; Blanco, Orestes; Departamento de Parasitología, Instituto de Medicina Tropical Pedro Kourí. La Habana, Cuba.; Llanos-Cuentas, Alejandro; Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia. Lima, Perú.; García, Ana L.; Universidad de San Simón. Cochabamba, Bolivia.; Valencia, Braulio M; Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia. Lima, Perú.; Muskus, Carlos; Programa de Estudio y Control de Enfermedades Tropicales, Universidad de Antioquia. Medellín, Colombia.; Van der Auwera, Gert; Biomedical Sciences Department. Institute of Tropical Medicine of Antwerp. Amberes, Bélgica.; Requena, José M; Centro de Biología Molecular Severo Ochoa. Madrid, España.


    Objectives. Explore a new target for molecular diagnosis of Leishmania. Materials and methods. We evaluated the utility of the gene that encodes the heat shock protein 20-kDa (Hsp20) for detecting Leishmania by polymerase chain reaction (PCR). PCR was normalized and analytical parameters were determined, as well as the validity and diagnostic accuracy, and concordance with the PCR - 18S. PCR-Hsp20 with DNA was obtained from a group of clinical samples from different sources. Results. The anal...

  19. Evolutionary origins of Hsp90 chaperones and a deep paralogy in their bacterial ancestors. (United States)

    Stechmann, Alexandra; Cavalier-Smith, Thomas


    The 82-90 kD family of molecular chaperone proteins has homologs in eukaryotes (Hsp90) and many eubacteria (HtpG) but not in Archaebacteria. We used representatives of all four different eukaryotic paralogs (cytosolic, endoplasmic reticulum (ER), chloroplast, mitochondrial) together with numerous eubacterial HtpG proteins for phylogenetic analyses to investigate their evolutionary origins. Our trees confirm that none of the organellar Hsp90s derives from the endosymbionts of early eukaryotes. Contrary to previous suggestions of distant origins through lateral gene transfer (LGT) all eukaryote Hsp90s are related to Gram-positive eubacterial HtpG proteins. The nucleocytosolic, ER and chloroplast Hsp90 paralogs are clearly mutually related. The origin of mitochondrial Hsp90 is more obscure, as these sequences are deeply nested within eubacteria. Our trees also reveal a deep split within eubacteria into a group of mainly long-branching sequences (including the eukaryote mitochondrial Hsp90s) and another group comprising exclusively short-branching HtpG proteins, from which the cytosolic/ER versions probably arose. Both versions are present in several eubacterial phyla, suggesting gene duplication very early in eubacterial evolution and multiple independent losses thereafter. We identified one probable case of LGT within eubacteria. However, multiple losses can simply explain the evolutionary pattern of the eubacterial HtpG paralogs and predominate over LGT. We suggest that the actinobacterial ancestor of eukaryotes harbored genes for both eubacterial HtpG paralogs, as the actinobacterium Streptomyces coelicolor still does; one could have given rise to the mitochondrial Hsp90 and the other, following another duplication event in the ancestral eukaryote, to the cytosolic and ER Hsp90 homologs.

  20. A Mycobacterium leprae Hsp65 mutant as a candidate for mitigating lupus aggravation in mice.

    Directory of Open Access Journals (Sweden)

    Eliana B Marengo

    Full Text Available Hsp60 is an abundant and highly conserved family of intracellular molecules. Increased levels of this family of proteins have been observed in the extracellular compartment in chronic inflammation. Administration of M. leprae Hsp65 [WT] in [NZBxNZW]F(1 mice accelerates the Systemic Lupus Erythematosus [SLE] progression whereas the point mutated K(409A Hsp65 protein delays the disease. Here, the biological effects of M. leprae Hsp65 Leader pep and K(409A pep synthetic peptides, which cover residues 352-371, are presented. Peptides had immunomodulatory effects similar to that observed with their respective proteins on survival and the combined administration of K(409A+Leader pep or K(409A pep+WT showed that the mutant forms were able to inhibit the deleterious effect of WT on mortality, indicating the neutralizing potential of the mutant molecules in SLE progression. Molecular modeling showed that replacing Lysine by Alanine affects the electrostatic potential of the 352-371 region. The number of interactions observed for WT is much higher than for Hsp65 K(409A and mouse Hsp60. The immunomodulatory effects of the point-mutated protein and peptide occurred regardless of the catalytic activity. These findings may be related to the lack of effect on survival when F(1 mice were inoculated with Hsp60 or K(409A pep. Our findings indicate the use of point-mutated Hsp65 molecules, such as the K(409A protein and its corresponding peptide, that may minimize or delay the onset of SLE, representing a new approach to the treatment of autoimmune diseases.

  1. Tah1 helix-swap dimerization prevents mixed Hsp90 co-chaperone complexes

    Energy Technology Data Exchange (ETDEWEB)

    Morgan, Rhodri M. L.; Pal, Mohinder; Roe, S. Mark; Pearl, Laurence H., E-mail:; Prodromou, Chrisostomos, E-mail: [University of Sussex, Falmer, Brighton BN1 9RQ (United Kingdom)


    A helix swap involving the fifth helix between two adjacently bound Tah1 molecules restores the normal binding environment of the conserved MEEVD peptide of Hsp90. Dimerization also explains how other monomeric TPR-domain proteins are excluded from forming inappropriate mixed co-chaperone complexes with Hsp90 and Tah1. Specific co-chaperone adaptors facilitate the recruitment of client proteins to the Hsp90 system. Tah1 binds the C-terminal conserved MEEVD motif of Hsp90, thus linking an eclectic set of client proteins to the R2TP complex for their assembly and regulation by Hsp90. Rather than the normal complement of seven α-helices seen in other tetratricopeptide repeat (TPR) domains, Tah1 unusually consists of the first five only. Consequently, the methionine of the MEEVD peptide remains exposed to solvent when bound by Tah1. In solution Tah1 appears to be predominantly monomeric, and recent structures have failed to explain how Tah1 appears to prevent the formation of mixed TPR domain-containing complexes such as Cpr6–(Hsp90){sub 2}–Tah1. To understand this further, the crystal structure of Tah1 in complex with the MEEVD peptide of Hsp90 was determined, which shows a helix swap involving the fifth α-helix between two adjacently bound Tah1 molecules. Dimerization of Tah1 restores the normal binding environment of the bound Hsp90 methionine residue by reconstituting a TPR binding site similar to that in seven-helix-containing TPR domain proteins. Dimerization also explains how other monomeric TPR-domain proteins are excluded from forming inappropriate mixed co-chaperone complexes.

  2. Study on HSP70 Gene Expression in Different Tissue of Cyprinus carpio%鲤鱼HSP70基因组织表达差异研究

    Institute of Scientific and Technical Information of China (English)

    林亚秋; 李瑞文


    [Objective] The aim of this study was to investigate whether HSP70 can be used as a stress monitoring indicator in Cyprinus carpio breeding. [Method] Based on HSP70 sequence of Cyprinus carpio (AY120894), one pair of primers was designed and synthesized, while the total RNA of liver tissues in Cyprinus carpio was extracted. Some cDNA fragments of Cyprinus carpio HSP70 were cloned by RT-PCR, and its differential expression in various tissues such as heart, intestine, mucus, gonad, swim bladder, gill and fin in Cyprinus carpio was also studied. [Result] The cDNA sequence of 480 bp was obtained from Cyprinus carpio HSP70 gene by RT-PCR amplification. Homology comparison between the deduced amino acid sequence after sequencing and that of other types of fish showed that the homology among Cyprinus carpio, Danio rerio, Ohcorhynehus mylciss, Paralichthys olivaceus, Xiphophoorus maculates and Carassius auratus was 96%, 98%, 98%, 96%, 96% and 96% respectively. The expression of HSP70 was detected in eight tissues of Cyprinus carpio. The expression was the highest in heart, followed by swim bladder and fin, but there was no significant difference between them (P>0.05). There was no significant difference among the expression in three tissues of intestine, mucus and fat (P>0.05), but their expression was significantly higher than those in gonad and gill (P<0.05). [Conclusion] HSP70 gene expression is a suitable criterion for monitoring the stress degree, stress capacity and healthy conditions in Cyprinus carpio breeding.

  3. Silencing of Hsp27 and Hsp72 in glioma cells as a tool for programmed cell death induction upon temozolomide and quercetin treatment

    Energy Technology Data Exchange (ETDEWEB)

    Jakubowicz-Gil, Joanna, E-mail: [Department of Comparative Anatomy and Anthropology, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin (Poland); Langner, Ewa [Department of Medical Biology, Institute of Agricultural Medicine, Jaczewskiego 2, 20-950 Lublin (Poland); Bądziul, Dorota [Department of Comparative Anatomy and Anthropology, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin (Poland); Wertel, Iwona [1st Department of Gynaecology, University School of Medicine, Staszica 16, 20-081 Lublin (Poland); Rzeski, Wojciech [Department of Medical Biology, Institute of Agricultural Medicine, Jaczewskiego 2, 20-950 Lublin (Poland); Department of Immunology and Virology, Maria Curie-Sklodowska University, Akademicka 19, 20-033 Lublin (Poland)


    The aim of the present study was to investigate whether silencing of Hsp27 or Hsp72 expression in glioblastoma multiforme T98G and anaplastic astrocytoma MOGGCCM cells increases their sensitivity to programmed cell death induction upon temozolomide and/or quercetin treatment. Transfection with specific siRNA was performed for the Hsp gene silencing. As revealed by microscopic observation and flow cytometry, the inhibition of Hsp expression was correlated with severe apoptosis induction upon the drug treatment studied. No signs of autophagy were detected. This was correlated with a decreased mitochondrial membrane potential, increased level of cytochrome c in the cytoplasm, and activation of caspase 3 and caspase 9. All these results suggest that the apoptotic signal was mediated by an internal pathway. Additionally, in a large percentage of cells treated with temozolomide, with or without quercetin, granules within the ER system were found, which was accompanied by an increased level of caspase 12 expression. This might be correlated with ER stress. Quercetin and temozolomide also changed the shape of nuclei from circular to “croissant like” in both transfected cell lines. Our results indicate that blocking of Hsp27 and Hsp72 expression makes T98G cells and MOGGCCM cells extremely vulnerable to apoptosis induction upon temozolomide and quercetin treatment and that programmed cell death is initiated by an internal signal. - Highlights: • Hsps gene silencing induced severe apoptosis upon temozolomide–quercetin treatment • Apoptosis in transfected glioma cells was initiated by internal signal • Programmed cell death was preceded by ER stress • Temozolomide–quercetin treatment changed nuclei shape in transfected glioma cells.

  4. An Impermeant Ganetespib Analog Inhibits Extracellular Hsp90-Mediated Cancer Cell Migration that Involves Lysyl Oxidase 2-like Protein

    Energy Technology Data Exchange (ETDEWEB)

    McCready, Jessica [Department of Natural Sciences, Assumption College, Worcester, MA 01609 (United States); Wong, Daniel S. [Department of Developmental Molecular and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111 (United States); Cell and Molecular Physiology Program, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, MA 02111 (United States); Burlison, Joseph A.; Ying, Weiwen [Synta Pharmaceuticals, Lexington, MA 02421 (United States); Jay, Daniel G., E-mail: [Department of Developmental Molecular and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111 (United States); Cell and Molecular Physiology Program, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, MA 02111 (United States)


    Extracellular Hsp90 (eHsp90) activates a number of client proteins outside of cancer cells required for migration and invasion. Therefore, eHsp90 may serve as a novel target for anti-metastatic drugs as its inhibition using impermeant Hsp90 inhibitors would not affect the numerous vital intracellular Hsp90 functions in normal cells. While some eHsp90 clients are known, it is important to establish other proteins that act outside the cell to validate eHsp90 as a drug target to limit cancer spread. Using mass spectrometry we identified two precursor proteins Galectin 3 binding protein (G3BP) and Lysyl oxidase 2-like protein (LOXL2) that associate with eHsp90 in MDA-MB231 breast cancer cell conditioned media and confirmed that LOXL2 binds to eHsp90 in immunoprecipitates. We introduce a novel impermeant Hsp90 inhibitor STA-12-7191 derived from ganetespib and show that it is markedly less toxic to cells and can inhibit cancer cell migration in a dose dependent manner. We used STA-12-7191 to test if LOXL2 and G3BP are potential eHsp90 clients. We showed that while LOXL2 can increase wound healing and compensate for STA-12-7191-mediated inhibition of wound closure, addition of G3BP had no affect on this assay. These findings support of role for LOXL2 in eHsp90 stimulated cancer cell migration and provide preliminary evidence for the use of STA-12-7191 to inhibit eHsp90 to limit cancer invasion.

  5. Molecular cloning and characterization of the MsHSP17.7 gene from Medicago sativa L. (United States)

    Li, Zhen-Yi; Long, Rui-Cai; Zhang, Tie-Jun; Yang, Qing-Chuan; Kang, Jun-Mei


    Heat shock proteins (HSPs) are ubiquitous protective proteins that play crucial roles in plant development and adaptation to stress, and the aim of this study is to characterize the HSP gene in alfalfa. Here we isolated a small heat shock protein gene (MsHSP17.7) from alfalfa by homology-based cloning. MsHSP17.7 contains a 477-bp open reading frame and encodes a protein of 17.70-kDa. The amino acid sequence shares high identity with MtHSP (93.98 %), PsHSP17.1 (83.13 %), GmHSP17.9 (74.10 %) and SlHSP17.6 (79.25 %). Phylogenetic analysis revealed that MsHSP17.7 belongs to the group of cytosolic class II small heat shock proteins (sHSP), and likely localizes to the cytoplasm. Quantitative RT-PCR indicated that MsHSP17.7 was induced by heat shock, high salinity, peroxide and drought stress. Prokaryotic expression indicated that the salt and peroxide tolerance of Escherichia coli was remarkably enhanced. Transgenic Arabidopsis plants overexpressing MsHSP17.7 exhibited increased root length of transgenic Arabidopsis lines under salt stress compared to the wild-type line. The malondialdehyde (MDA) levels in the transgenic lines were significantly lower than in wild-type, although proline levels were similar between transgenic and wild-type lines. MsHSP17.7 was induced by heat shock, high salinity, oxidative stress and drought stress. Overexpression analysis suggests that MsHSP17.7 might play a key role in response to high salinity stress.

  6. BAG2 Interferes with CHIP-Mediated Ubiquitination of HSP72. (United States)

    Schönbühler, Bianca; Schmitt, Verena; Huesmann, Heike; Kern, Andreas; Gamerdinger, Martin; Behl, Christian


    The maintenance of cellular proteostasis is dependent on molecular chaperones and protein degradation pathways. Chaperones facilitate protein folding, maturation, and degradation, and the particular fate of a misfolded protein is determined by the interaction of chaperones with co-chaperones. The co-factor CHIP (C-terminus of HSP70-inteacting protein, STUB1) ubiquitinates chaperone substrates and directs proteins to the cellular degradation systems. The activity of CHIP is regulated by two co-chaperones, BAG2 and HSPBP1, which are potent inhibitors of the E3 ubiquitin ligase activity. Here, we examined the functional correlation of HSP72, CHIP, and BAG2, employing human primary fibroblasts. We showed that HSP72 is a substrate of CHIP and that BAG2 efficiently prevented the ubiquitination of HSP72 in young cells as well as aged cells. Aging is associated with a decline in proteostasis and we observed increased protein levels of CHIP as well as BAG2 in senescent cells. Interestingly, the ubiquitination of HSP72 was strongly reduced during aging, which revealed that BAG2 functionally counteracted the increased levels of CHIP. Interestingly, HSPBP1 protein levels were down-regulated during aging. The data presented here demonstrates that the co-chaperone BAG2 influences HSP72 protein levels and is an important modulator of the ubiquitination activity of CHIP in young as well as aged cells.

  7. Variation in Hsp70-1A Expression Contributes to Skin Color Diversity. (United States)

    Murase, Daiki; Hachiya, Akira; Fullenkamp, Rachel; Beck, Anita; Moriwaki, Shigeru; Hase, Tadashi; Takema, Yoshinori; Manga, Prashiela


    The wide range in human skin color results from varying levels of the pigment melanin. Genetic mechanisms underlying coloration differences have been explored, but identified genes do not account for all variation seen in the skin color spectrum. Post-transcriptional and post-translational regulation of factors that determine skin color, including melanin synthesis in epidermal melanocytes, melanosome transfer to keratinocytes, and melanosome degradation, is also critical for pigmentation. We therefore investigated proteins that are differentially expressed in melanocytes derived from either white or African American skin. Two-dimensional gel electrophoresis and mass spectrometry demonstrated that heat shock protein 70-1A (Hsp70-1A) protein levels were significantly higher in African American melanocytes compared with white melanocytes. Hsp70-1A expression significantly correlated with levels of tyrosinase, the rate-limiting melanogenic enzyme, consistent with a proposed role for Hsp70 family members in tyrosinase post-translational modification. In addition, pharmacologic inhibition and small interfering RNA-mediated downregulation of Hsp70-1A correlated with pigmentation changes in cultured melanocytes, modified human skin substitutes, and ex vivo skin. Furthermore, Hsp70-1A inhibition led to increased autophagy-mediated melanosome degradation in keratinocytes. Our data thus reveal that epidermal Hsp70-1A contributes to the diversity of skin color by regulating the amount of melanin synthesized in melanocytes and modulating autophagic melanosome degradation in keratinocytes.

  8. Immunohistochemistry of human Hsp60 in health and disease: from autoimmunity to cancer. (United States)

    Cappello, Francesco; de Macario, Everly Conway; Zummo, Giovanni; Macario, Alberto J L


    Hsp60 (also called Cpn60) is a chaperonin with essential functions for cell physiology and survival. Additionally, its involvement in the pathogenesis of a number of diseases (e.g., some autoimmune disorders and cancer) is becoming evident with new research. For example, the distribution and levels of Hsp60 in cells and tissues have been found altered in many pathologic conditions, and the significance of these alterations is being investigated in a number of laboratories. The aim of this ongoing research is to determine the meaning of these Hsp60 alterations with regard to pathogenetic mechanisms, diagnosis, classification of lesions, and assessing of prognosis and response to treatment. Hsp60 occurs in the mitochondria, i.e., its typical residence according to classic knowledge, and also in other locales, such as the cytosol, the cell membrane, the intercellular space, and biological fluids (e.g., blood and cerebrospinal fluid). Detection and quantitative determinations in all these locations are becoming essential components of laboratory pathology in clinics and research. Consequently, immunohistochemistry targeting Hsp60 is also becoming essential for pathologists and researchers interested in disorders involving this chaperonin. In this chapter, we briefly summarize some recent discoveries on the participation of Hsp60 in the pathogenesis of human diseases and describe in detail how to perform immunohistochemical reactions for detecting the chaperonin, determining its location, and measuring its levels of expression.

  9. Expression of Hsp70 and Caspase-3 in rabbits after severe traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jing; TAO Dai-qin; ZHAO Hui; YIN Zhi-yong


    Objective:To investigate the expression of Caspase-3 and Hsp70 in rabbits after severe traumatic brain injury (TBI) and to explore the feasibility of its application in estimation of injury time in forensic medicine.Methods:A rabbit model of heavy TBI was developed by high velocity impact on the parietal bone with an iron stick.Totally 8 healthy adult New Zealand white rabbits were randomly divided into control group (n=2) and injury group (n=6).Four hours after injury,tissue specimens from the parietal lobe,temporal lobe,occipital lobe,cerebellum and brainstem were harvested to detect the expression of Hsp70 and Caspase-3 by immunohistochemistry.Besides,the gray values of cells positive for Hsp70 and Caspase-3 were analyzed with an image analyzer.Results:Immunohistochemistry staining demonstrated a low level of Caspase-3 and Hsp70 expression in normal control group.While in injury group,both the Caspase-3and Hsp70 expression was significantly elevated (P<0.05).Positive cells gathered around the lesion focus.Occipital lobe and cerebellum had fewer positive cells while temporal and brainstem had the fewest.Conclusion:The expression of Caspase-3 and Hsp70 at an early stage following severe TBI is characteristic and can be applied to estimate the time of injury.

  10. Little effect of HSP90 inhibition on the quantitative wing traits variation in Drosophila melanogaster. (United States)

    Takahashi, Kazuo H


    Drosophila wings have been a model system to study the effect of HSP90 on quantitative trait variation. The effect of HSP90 inhibition on environmental buffering of wing morphology varies among studies while the genetic buffering effect of it was examined in only one study and was not detected. Variable results so far might show that the genetic background influences the environmental and genetic buffering effect of HSP90. In the previous studies, the number of the genetic backgrounds used is limited. To examine the effect of HSP90 inhibition with a larger number of genetic backgrounds than the previous studies, 20 wild-type strains of Drosophila melanogaster were used in this study. Here I investigated the effect of HSP90 inhibition on the environmental buffering of wing shape and size by assessing within-individual and among-individual variations, and as a result, I found little or very weak effects on environmental and genetic buffering. The current results suggest that the role of HSP90 as a global regulator of environmental and genetic buffering is limited at least in quantitative traits.

  11. Topically Applied Hsp90 Blocker 17AAG Inhibits Autoantibody-Mediated Blister-Inducing Cutaneous Inflammation. (United States)

    Tukaj, Stefan; Bieber, Katja; Kleszczyński, Konrad; Witte, Mareike; Cames, Rebecca; Kalies, Kathrin; Zillikens, Detlef; Ludwig, Ralf J; Fischer, Tobias W; Kasperkiewicz, Michael


    Cell stress-inducible Hsp90 has been recognized as key player in mediating inflammatory responses. Although its systemic blockade was successfully used to treat autoimmune diseases in preclinical models, efficacy of a topical route of Hsp90 inhibitor administration has so far not been evaluated in chronic inflammatory and autoimmune-mediated dermatoses. Here, effects of the Hsp90 blocker 17-allylamino-demethoxygeldanamycin (17AAG) applied topically to the skin were determined in experimental inflammatory epidermolysis bullosa acquisita (EBA), an anti-type VII collagen autoantibody-induced blistering skin disease. Topical 17AAG ameliorated clinical disease severity when given before or during the occurrence of skin lesions without causing cutaneous or systemic toxicity in mice with antibody transfer- and immunization-induced EBA. In both EBA models and in the setting of locally induced inflammation, topical 17AAG treatment was associated with (i) reduced neutrophilic infiltrates, (ii) decreased NF-κB activation, (iii) lowered expression of matrix metalloproteinases and Flii, and (iv) induction of anti-inflammatory Hsp70 in the skin. Our results suggest that topical delivery of Hsp90 antagonists, offering the benefit of a reduced risk of systemic adverse effects of Hsp90 inhibition, may be useful for the control of EBA and possibly other related inflammatory skin disorders.

  12. The evolutionary capacitor HSP90 buffers the regulatory effects of mammalian endogenous retroviruses. (United States)

    Hummel, Barbara; Hansen, Erik C; Yoveva, Aneliya; Aprile-Garcia, Fernando; Hussong, Rebecca; Sawarkar, Ritwick


    Understanding how genotypes are linked to phenotypes is important in biomedical and evolutionary studies. The chaperone heat-shock protein 90 (HSP90) buffers genetic variation by stabilizing proteins with variant sequences, thereby uncoupling phenotypes from genotypes. Here we report an unexpected role of HSP90 in buffering cis-regulatory variation affecting gene expression. By using the tripartite-motif-containing 28 (TRIM28; also known as KAP1)-mediated epigenetic pathway, HSP90 represses the regulatory influence of endogenous retroviruses (ERVs) on neighboring genes that are critical for mouse development. Our data based on natural variations in the mouse genome show that genes respond to HSP90 inhibition in a manner dependent on their genomic location with regard to strain-specific ERV-insertion sites. The evolutionary-capacitor function of HSP90 may thus have facilitated the exaptation of ERVs as key modifiers of gene expression and morphological diversification. Our findings add a new regulatory layer through which HSP90 uncouples phenotypic outcomes from individual genotypes.

  13. HSP90 promotes Burkitt lymphoma cell survival by maintaining tonic B-cell receptor signaling. (United States)

    Walter, Roland; Pan, Kuan-Ting; Doebele, Carmen; Comoglio, Federico; Tomska, Katarzyna; Bohnenberger, Hanibal; Young, Ryan M; Jacobs, Laura; Keller, Ulrich; Bönig, Halvard; Engelke, Michael; Rosenwald, Andreas; Urlaub, Henning; Staudt, Louis M; Serve, Hubert; Zenz, Thorsten; Oellerich, Thomas


    Burkitt lymphoma (BL) is an aggressive B-cell neoplasm that is currently treated by intensive chemotherapy in combination with anti-CD20 antibodies. Because of their toxicity, current treatment regimens are often not suitable for elderly patients or for patients in developing countries where BL is endemic. Targeted therapies for BL are therefore needed. In this study, we performed a compound screen in 17 BL cell lines to identify small molecule inhibitors affecting cell survival. We found that inhibitors of heat shock protein 90 (HSP90) induced apoptosis in BL cells in vitro at concentrations that did not affect normal B cells. By global proteomic and phosphoproteomic profiling, we show that, in BL, HSP90 inhibition compromises the activity of the pivotal B-cell antigen receptor (BCR)-proximal effector spleen tyrosine kinase (SYK), which we identified as an HSP90 client protein. Consistently, expression of constitutively active TEL-SYK counteracted the apoptotic effect of HSP90 inhibition. Together, our results demonstrate that HSP90 inhibition impairs BL cell survival by interfering with tonic BCR signaling, thus providing a molecular rationale for the use of HSP90 inhibitors in the treatment of BL.

  14. Expression dynamics of HSP70 during chronic heat stress in Tharparkar cattle (United States)

    Bharati, Jaya; Dangi, S. S.; Chouhan, V. S.; Mishra, S. R.; Bharti, M. K.; Verma, V.; Shankar, O.; Yadav, V. P.; Das, K.; Paul, A.; Bag, S.; Maurya, V. P.; Singh, G.; Kumar, P.; Sarkar, M.


    Six male Tharparkar cattle aged 2-3 years were selected for the study. The animals were acclimatized in the psychrometric chamber at thermoneutral zone (TNZ) for 15 days and then exposed to 42 °C temperature up to 23 days followed by 12 days of recovery period. Physiological responses were estimated, and peripheral blood mononuclear cells (PBMCs) were isolated at TNZ on day 1, day 5, and day 12; after 6 h of heat stress exposure on day 16 to day 20, day 25, day 30, day 32, day 34, day 36, and day 38; and a recovery period on day 45 and day 50. The PBMCs were cultured to study the effect of thermal challenge on HSP70 messenger RNA (mRNA) expression pattern at different temperature-time combinations. The mRNA and protein expression of HSP70 in PBMCs along with serum extracellular HSP70 (eHSP70) was increased (P heat stress challenge treatment as compared to control in cultured PBMCs. HSP70 expression was found to be higher (P heat exposure (corresponds to chronic heat stress) as compared to the first 5 days of heat stress (corresponds to short-term heat stress) and control period at TNZ. The present findings indicate that HSP70 is possibly involved in heat stress adaptive response in Tharparkar cattle and the biphasic expression pattern may be providing a second window of protection during chronic heat stress.

  15. The crystal structure of the Hsp90 co-chaperone Cpr7 from Saccharomyces cerevisiae. (United States)

    Qiu, Yu; Ge, Qiangqiang; Wang, Mingxing; Lv, Hui; Ebrahimi, Mohammad; Niu, Liwen; Teng, Maikun; Li, Xu


    The versatility of Hsp90 can be attributed to the variety of co-chaperone proteins that modulate the role of Hsp90 in many cellular processes. As a co-chaperone of Hsp90, Cpr7 is essential for accelerating the cell growth in an Hsp90-containing trimeric complex. Here, we report the crystal structure of Cpr7 at a resolution of 1.8Å. It consists of an N-terminal PPI domain and a C-terminal TPR domain, and exhibits a U-shape conformation. Our studies revealed the aggregation state of Cpr7 in solution and the interaction properties between Cpr7 and the MEEVD sequence from the C-terminus of Hsp90. In addition, the structure and sequence analysis between Cpr7 and homologues revealed the structure basis both for the function differences between Cpr6 and Cpr7 and the functional complements between Cns1 and Cpr7. Our studies facilitate the understanding of Cpr7 and provide decent insights into the molecular mechanisms of the Hsp90 co-chaperone pathway.

  16. Mechanistic basis for the recognition of a misfolded protein by the molecular chaperone Hsp90. (United States)

    Oroz, Javier; Kim, Jin Hae; Chang, Bliss J; Zweckstetter, Markus


    The critical toxic species in over 40 human diseases are misfolded proteins. Their interaction with molecular chaperones such as Hsp90, which preferentially interacts with metastable proteins, is essential for the blocking of disease progression. Here we used nuclear magnetic resonance (NMR) spectroscopy to determine the three-dimensional structure of the misfolded cytotoxic monomer of the amyloidogenic human protein transthyretin, which is characterized by the release of the C-terminal β-strand and perturbations of the A-B loop. The misfolded transthyretin monomer, but not the wild-type protein, binds to human Hsp90. In the bound state, the Hsp90 dimer predominantly populates an open conformation, and transthyretin retains its globular structure. The interaction surface for the transthyretin monomer comprises the N-terminal and middle domains of Hsp90 and overlaps with that of the Alzheimer's-disease-related protein tau. Taken together, the data suggest that Hsp90 uses a mechanism for the recognition of aggregation-prone proteins that is largely distinct from those of other Hsp90 clients.

  17. J-dot targeting of an exported HSP40 in Plasmodium falciparum-infected erythrocytes. (United States)

    Petersen, Wiebke; Külzer, Simone; Engels, Sonja; Zhang, Qi; Ingmundson, Alyssa; Rug, Melanie; Maier, Alexander G; Przyborski, Jude M


    Plasmodium falciparum exports a large number of proteins to its host cell, the mature human erythrocyte, where they are involved in host cell modification. Amongst the proteins trafficked to the host cell, many are heat shock protein (HSP)40 homologues. We previously demonstrated that at least two exported PfHSP40s (referred to as PFE55 and PFA660) localise to mobile structures in the P. falciparum-infected erythrocyte (Kulzer et al., 2010), termed J-dots. The complete molecular content of these structures has not yet been completely resolved, however it is known that they also contain an exported HSP70, PfHSP70x, and are potentially involved in transport of the major cytoadherance ligand, PfEMP1, through the host cell. To understand more about the nature of the association of exported HSP40s with J-dots, here we have studied the signal requirements for recruitment of the proteins to these structures. By expressing various exported GFP chimeras, we can demonstrate that the predicted substrate binding domain is necessary and sufficient for J-dot targeting. This targeting only occurs in human erythrocytes infected with P. falciparum, as it is not conserved when expressing a P. falciparum HSP40 in Plasmodium berghei-infected murine red blood cells, suggesting that J-dots are P. falciparum-specific. This data reveals a new mechanism for targeting of exported proteins to intracellular structures in the P. falciparum-infected erythrocyte.

  18. DNA double strand breaks and Hsp70 expression in proton irradiated living cells

    Energy Technology Data Exchange (ETDEWEB)

    Fiedler, Anja [Institute for Experimental Physics II, University of Leipzig (Germany) and Faculty of Biology, Pharmacy and Psychology, University of Leipzig (Germany)]. E-mail:; Reinert, Tilo [Institute for Experimental Physics II, University of Leipzig (Germany); Tanner, Judith [Clinic and Polyclinic for Radiation Oncology, University of Halle-Wittenberg (Germany); Butz, Tilman [Institute for Experimental Physics II, University of Leipzig (Germany)


    DNA double strand breaks (DSBs) in living cells can be directly provoked by ionising radiation. DSBs can be visualized by immunostaining the phosphorylated histone {gamma}H2AX. Our concern was to test the feasibility of {gamma}H2AX staining for a direct visualization of single proton hits. If single protons produce detectable foci, DNA DSBs could be used as 'biological track detectors' for protons. Ionising radiation can also damage proteins indirectly by inducing free radicals. Heat shock proteins (Hsp) help to refold or even degrade the damaged proteins. The level of the most famous heat shock protein Hsp70 is increased by ionising radiation. We investigated the expression of {gamma}H2AX and Hsp70 after cross and line patterned irradiation with counted numbers of 2.25 MeV protons on primary human skin fibroblasts. The proton induced DSBs appear more delocalised than it was expected by the ion hit accuracy. Cooling the cells before the irradiation reduces the delocalisation of DNA DSBs, which is probably caused by the reduced diffusion of DNA damaging agents. Proton irradiation seems to provoke protein damages mainly in the cytoplasm indicated by cytoplasmic Hsp70 aggregates. On the contrary, in control heat shocked cells the Hsp70 was predominantly localized in the cell nucleus. However, the irradiated area could not be recognized, all cells on the Si{sub 3}N{sub 4} window showed a homogenous Hsp70 expression pattern.

  19. Small heat shock protein 20 (HspB6) in cardiac hypertrophy and failure. (United States)

    Fan, Guo-Chang; Kranias, Evangelia G


    Hsp20, referred to as HspB6, is constitutively expressed in various tissues. Specifically, HspB6 is most highly expressed in different types of muscle including vascular, airway, colonic, bladder, and uterine smooth muscle; cardiac muscle; and skeletal muscle. It can be phosphorylated at Ser-16 by both cAMP- and cGMP-dependent protein kinases (PKA/PKG). Recently, Hsp20 and its phosphorylation have been implicated in multiple physiological and pathophysiological processes including smooth muscle relaxation, platelet aggregation, exercise training, myocardial infarction, atherosclerosis, insulin resistance and Alzheimer's disease. In the heart, key advances have been made in elucidating the significance of Hsp20 in contractile function and cardioprotection over the last decade. This mini-review highlights exciting findings in animal models and human patients, with special emphasis on the potential salutary effects of Hsp20 in heart disease. This article is part of a special issue entitled "Key Signaling Molecules in Hypertrophy and Heart Failure."

  20. BioMagResBank.

    NARCIS (Netherlands)

    Ulrich, E.L.; Akutsu, H.; Doreleijers, J.; Harano, Y.; Ioannidis, Y.E.; Lin, J.; Livny, M.; Mading, S.; Maziuk, D.; Miller, Z.; Nakatani, E.; Schulte, C.F.; Tolmie, D.E.; Wenger, R Kent; Yao, H.; Markley, J.L.


    The BioMagResBank (BMRB: is a repository for experimental and derived data gathered from nuclear magnetic resonance (NMR) spectroscopic studies of biological molecules. BMRB is a partner in the Worldwide Protein Data Bank (wwPDB). The BMRB archive consists of four main data deposi

  1. Chemical modulators of heat shock protein 70 (Hsp70) by sequential, microwave-accelerated reactions on solid phase. (United States)

    Wisén, Susanne; Androsavich, John; Evans, Christopher G; Chang, Lyra; Gestwicki, Jason E


    Molecular chaperones, such as Hsp70 and Hsp90, are responsible for a variety of protective, anti-apoptotic functions. While inhibitors of Hsp90, such as geldanamycin and its derivative 17-AAG, are well known and important anti-cancer leads, Hsp70 has received less attention. Interesting lead candidates for Hsp70 share a dihydropyrimidine core; however, the preferred display of pendant functionality is still not clear. Here, we take advantage of the versatility of peptides to explore the requirements for activity. An exploratory compound collection was assembled by performing a Biginelli cyclocondensation at the terminus of a resin-bound beta-peptide. Liberation from solid support yielded peptide-modified dihydropyrimidines and, within this series, we uncovered compounds that alter the ATPase activity of Hsp70 and its bacterial ortholog, DnaK. Moreover, we identified important contributions made by aromatic, hydrophobic groups. These chemical probes could be used to study the roles of this molecular chaperone in disease.

  2. A Hyperactive Signalosome in Acute Myeloid Leukemia Drives Addiction to a Tumor-Specific Hsp90 Species

    Directory of Open Access Journals (Sweden)

    Hongliang Zong


    Full Text Available Acute myeloid leukemia (AML is a heterogeneous and fatal disease with an urgent need for improved therapeutic regimens given that most patients die from relapsed disease. Irrespective of mutation status, the development of aggressive leukemias is enabled by increasing dependence on signaling networks. We demonstrate that a hyperactive signalosome drives addiction of AML cells to a tumor-specific Hsp90 species (teHsp90. Through genetic, environmental, and pharmacologic perturbations, we demonstrate a direct and quantitative link between hyperactivated signaling pathways and apoptotic sensitivity of AML to teHsp90 inhibition. Specifically, we find that hyperactive JAK-STAT and PI3K-AKT signaling networks are maintained by teHsp90 and, in fact, gradual activation of these networks drives tumors increasingly dependent on teHsp90. Thus, although clinically aggressive AML survives via signalosome activation, this addiction creates a vulnerability that can be exploited with Hsp90-directed therapy.

  3. Cloning and characterization of a heat shock protein 70 gene, MsHSP70-1, in Medicago sativa

    Institute of Scientific and Technical Information of China (English)

    Zhishui He; Rong Xie; Yanzhang Wang; Huasong Zou; Jiabi Zhu; Guanqiao Yu


    Some members of the heat shock protein 70 (HSP70) family have important functions in organism development. Here, we identified an alfalfa (Medicago sativa L. cv. Algonquin) HSP70 gene, MsHSP70-1, using cDNA array and reverse transcription-polymerase chain reaction. This gene contains a 1947 bp open reading frame encoding a protein of 649 amino acids.This protein contains all conserved domains,motifs.and characteristic sequences of plant HSP70s.The expression of MsHsP70-1 is enhanced in nodule compared withroot,stem,leaf,and flower,and throughout the process of nodule development.Northern hybridization analysis indicated that the expression of MsHSP70-1 in nodule requires the active bacA gene of rhizobia.These results suggested that MsHSP70-1 might play an important role in alfalfa nodule development.

  4. HSP47与宫腔粘连的关系及在纤维化疾病中的意义%The Relationship between HSP47 and Intrauterine Adhesions and Its Significance in Fibrosis Diseases

    Institute of Scientific and Technical Information of China (English)

    王祥珍; 马娜娜


    Objective:To review the significance of HSP47 in fibrosis disease and explore the pathogenesis of intrauterine adhesions. Methods:The significance of HSP47 in fibrosis diseases such as liver fibrosis,pulmonary fibrosis,renal fibrosis and skin scar was discussed,and the relationship between HSP47 and intrauterine adhesions was explored. Results:HSP47 was high expression in fibrosis disease,and there was relationship between HSP47 and intrauterine adhesions. Conclusion:The relationship between HSP47 and intrauterine adhesion are discussed.%目的:综述HSP47在纤维化疾病中的意义,探索宫腔粘连的发病机制。方法:对HSP47在肝纤维化、肺纤维化、肾纤维化、皮肤瘢痕等纤维化疾病中的意义进行探讨,探索其与宫腔粘连的关系。结果:HSP47在纤维化疾病中高表达,与宫腔粘连的发生存在某种关系。结论:对宫腔粘连的发病机制进行探讨和展望。

  5. Influence of NaCl, urea, potassium and pH on HSP72 expression in MDCK cells. (United States)

    Neuhofer, W; Müller, E; Grünbein, R; Thurau, K; Beck, F X


    The renal inner medulla is characterised by elevated extracellular concentrations of NaCl, urea, potassium and hydrogen ions, an environment that may affect cell viability negatively. High amounts of HSP72, a stress protein allowing cells to resist harmful situations, are also observed in this region. The present study examined HSP72 induction by various medullary stress factors, individually or in combination, in MDCK cells, a renal epithelial cell line expressing characteristics of the medullary collecting duct. MDCK cells were incubated for 3 days in media containing elevated concentrations of NaCl, urea, potassium and hydrogen ions individually or in combination. HSP72 mRNA and protein expression were determined by Northern and Western blot analyses, respectively. HSP72 expression was enhanced moderately by addition of 50 mM NaCl to normal medium at pH 7.4 but enhanced strongly when added at pH 6.5. The latter degree of HSP72 induction was comparable to that observed when 150 mM NaCl was added at pH 7.4. In normal medium (pH 7.4) containing 300 mM urea, MDCK HSP72 expression was not different from controls. In contrast, urea-induced HSP72 expression was clearly evident when medium pH was lowered to 6.5. Potassium at 20 or 40 mM induced HSP72 only slightly. These results indicate that expression of HSP72 in renal epithelial cells is regulated synergistically by NaCl, urea and pH. Since HSP72 is only slightly induced by increased potassium, this probably reflects the changes in medium osmolality rather than a specific effect of potassium. The high medullary HSP72 content observed even in diuresis may be due to co-operative effects of medullary solutes on HSP72 expression.

  6. Interaction between Hsp60 and Bax in normal human myocardium and in myocardium affected by dilated cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Tykhonkova I. O.


    Full Text Available The main functional compartments of molecular chaperone Hsp60 are mitochondria and cytoplasm. Up to 30 % of Hsp60 are located in cytoplasm of cardiomyocytes. The interaction between molecular chaperone Hsp60 and proapoptotic Bax protein in the cytoplasmic fraction from normal human heart tissue has been revealed by co-immunoprecipitation in contrast to myocardium affected by dilated cardiomyopathy, where this interaction has not been observed

  7. The expression and function of hsp30-like small heat shock protein genes in amphibians, birds, fish, and reptiles. (United States)

    Heikkila, John J


    Small heat shock proteins (sHSPs) are a superfamily of molecular chaperones with important roles in protein homeostasis and other cellular functions. Amphibians, reptiles, fish and birds have a shsp gene called hsp30, which was also referred to as hspb11 or hsp25 in some fish and bird species. Hsp30 genes, which are not found in mammals, are transcribed in response to heat shock or other stresses by means of the heat shock factor that is activated in response to an accumulation of unfolded protein. Amino acid sequence analysis revealed that representative HSP30s from different classes of non-mammalian vertebrates were distinct from other sHSPs including HSPB1/HSP27. Studies with amphibian and fish recombinant HSP30 determined that they were molecular chaperones since they inhibited heat- or chemically-induced aggregation of unfolded protein. During non-mammalian vertebrate development, hsp30 genes were differentially expressed in selected tissues. Also, heat shock-induced stage-specific expression of hsp30 genes in frog embryos was regulated at the level of chromatin structure. In adults and/or tissue culture cells, hsp30 gene expression was induced by heat shock, arsenite, cadmium or proteasomal inhibitors, all of which enhanced the production of unfolded/damaged protein. Finally, immunocytochemical analysis of frog and chicken tissue culture cells revealed that proteotoxic stress-induced HSP30 accumulation co-localized with aggresome-like inclusion bodies. The congregation of damaged protein in aggresomes minimizes the toxic effect of aggregated protein dispersed throughout the cell. The current availability of probes to detect the presence of hsp30 mRNA or encoded protein has resulted in the increased use of hsp30 gene expression as a marker of proteotoxic stress in non-mammalian vertebrates.

  8. Convergent sets of data from in vivo and in vitro methods point to an active role of Hsp60 in chronic obstructive pulmonary disease pathogenesis.

    Directory of Open Access Journals (Sweden)

    Francesco Cappello

    Full Text Available BACKGROUND: It is increasingly clear that some heat shock proteins (Hsps play a role in inflammation. Here, we report results showing participation of Hsp60 in the pathogenesis of chronic obstructive pulmonary diseases (COPD, as indicated by data from both in vivo and in vitro analyses. METHODS AND RESULTS: Bronchial biopsies from patients with stable COPD, smoker controls with normal lung function, and non-smoker controls were studied. We quantified by immunohistochemistry levels of Hsp10, Hsp27, Hsp40, Hsp60, Hsp70, Hsp90, and HSF-1, along with levels of inflammatory markers. Hsp10, Hsp40, and Hsp60 were increased during progression of disease. We found also a positive correlation between the number of neutrophils and Hsp60 levels. Double-immunostaining showed that Hsp60-positive neutrophils were significantly increased in COPD patients. We then investigated in vitro the effect on Hsp60 expression in bronchial epithelial cells (16HBE caused by oxidative stress, a hallmark of COPD mucosa, which we induced with H₂O₂. This stressor determined increased levels of Hsp60 through a gene up-regulation mechanism involving NFkB-p65. Release of Hsp60 in the extracellular medium by the bronchial epithelial cells was also increased after H₂O₂ treatment in the absence of cell death. CONCLUSIONS: This is the first report clearly pointing to participation of Hsps, particularly Hsp60, in COPD pathogenesis. Hsp60 induction by NFkB-p65 and its release by epithelial cells after oxidative stress can have a role in maintaining inflammation, e.g., by stimulating neutrophils activity. The data open new scenarios that might help in designing efficacious anti-inflammatory therapies centered on Hsp60 and applicable to COPD.

  9. Convergent Sets of Data from In Vivo and In Vitro Methods Point to an Active Role of Hsp60 in Chronic Obstructive Pulmonary Disease Pathogenesis (United States)

    Campanella, Claudia; Vicari, Chiara; Gnemmi, Isabella; Zanini, Andrea; Spanevello, Antonio; Capelli, Armando; La Rocca, Giampiero; Anzalone, Rita; Bucchieri, Fabio; D'Anna, Silvestro Ennio; Ricciardolo, Fabio L. M.; Brun, Paola; Balbi, Bruno; Carone, Mauro; Zummo, Giovanni; de Macario, Everly Conway; Macario, Alberto J. L.; Di Stefano, Antonino


    Background It is increasingly clear that some heat shock proteins (Hsps) play a role in inflammation. Here, we report results showing participation of Hsp60 in the pathogenesis of chronic obstructive pulmonary diseases (COPD), as indicated by data from both in vivo and in vitro analyses. Methods and Results Bronchial biopsies from patients with stable COPD, smoker controls with normal lung function, and non-smoker controls were studied. We quantified by immunohistochemistry levels of Hsp10, Hsp27, Hsp40, Hsp60, Hsp70, Hsp90, and HSF-1, along with levels of inflammatory markers. Hsp10, Hsp40, and Hsp60 were increased during progression of disease. We found also a positive correlation between the number of neutrophils and Hsp60 levels. Double-immunostaining showed that Hsp60-positive neutrophils were significantly increased in COPD patients. We then investigated in vitro the effect on Hsp60 expression in bronchial epithelial cells (16HBE) caused by oxidative stress, a hallmark of COPD mucosa, which we induced with H2O2. This stressor determined increased levels of Hsp60 through a gene up-regulation mechanism involving NFkB-p65. Release of Hsp60 in the extracellular medium by the bronchial epithelial cells was also increased after H2O2 treatment in the absence of cell death. Conclusions This is the first report clearly pointing to participation of Hsps, particularly Hsp60, in COPD pathogenesis. Hsp60 induction by NFkB-p65 and its release by epithelial cells after oxidative stress can have a role in maintaining inflammation, e.g., by stimulating neutrophils activity. The data open new scenarios that might help in designing efficacious anti-inflammatory therapies centered on Hsp60 and applicable to COPD. PMID:22140545

  10. New phagotrophic euglenoid species (new genus Decastava; Scytomonas saepesedens; Entosiphon oblongum), Hsp90 introns, and putative euglenoid Hsp90 pre-mRNA insertional editing. (United States)

    Cavalier-Smith, Thomas; Chao, Ema E; Vickerman, Keith


    We describe three new phagotrophic euglenoid species by light microscopy and 18S rDNA and Hsp90 sequencing: Scytomonas saepesedens; Decastava edaphica; Entosiphon oblongum. We studied Scytomonas and Decastava ultrastructure. Scytomonas saepesedens feeds when sessile with actively beating cilium, and has five pellicular strips with flush joints and Calycimonas-like microtubule-supported cytopharynx. Decastava, sister to Keelungia forming new clade Decastavida on 18S rDNA trees, has 10 broad strips with cusp-like joints, not bifurcate ridges like Ploeotia and Serpenomonas (phylogenetically and cytologically distinct genera), and Serpenomonas-like feeding apparatus (8-9 unreinforced microtubule pairs loop from dorsal jaw support to cytostome). Hsp90 and 18S rDNA trees group Scytomonas with Petalomonas and show Entosiphon as the earliest euglenoid branch. Basal euglenoids have rigid longitudinal strips; derived clade Spirocuta has spiral often slideable strips. Decastava Hsp90 genes have introns. Decastava/Entosiphon Hsp90 frameshifts imply insertional RNA editing. Petalomonas is too heterogeneous in pellicle structure for one genus; we retain Scytomonas (sometimes lumped with it) and segregate four former Petalomonas as new genus Biundula with pellicle cross section showing 2-8 smooth undulations and typified by Biundula (=Petalomonas) sphagnophila comb. n. Our taxon-rich site-heterogeneous rDNA trees confirm that Heteronema is excessively heterogeneous; therefore we establish new genus Teloprocta for Heteronema scaphurum.

  11. Stress Proteins (hsp70, hsp60) Induced in Isopods and Nematodes by Field Exposure to Metals in a Gradient near Avonmouth, UK

    NARCIS (Netherlands)

    Arts, M.S.J.; Schill, R.O.; Knigge, T.; Eckwert, H.; Kammenga, J.E.; Köhler, H.R.


    Heat shock proteins (hsps) are potential biomarkers for monitoring environmental pollution. In this study, the use of hsps as biomarkers in field bioassays was evaluated in terrestrial invertebrates exposed to a metal gradient near Avonmouth, UK. We investigated the hsp70 response in resident and tr

  12. Molecular cloning and functional analysis of the drought tolerance gene MsHSP70 from alfalfa (Medicago sativa L.). (United States)

    Li, Zhenyi; Long, Ruicai; Zhang, Tiejun; Wang, Zhen; Zhang, Fan; Yang, Qingchuan; Kang, Junmei; Sun, Yan


    Heat shock proteins (HSPs) are a ubiquitously expressed class of protective proteins that play a key role in plant response to stressful conditions. This study aimed to characterize and investigate the function of an HSP gene in alfalfa (Medicago sativa). MsHSP70, which contains a 2028-bp open reading frame, was identified through homology cloning. MsHSP70 shares high sequence identity (94.47%) with HSP70 from Medicago truncatula. Expression analysis of MsHSP70 in alfalfa organs revealed a relatively higher expression level in aerial organs such as flowers, stems and leaves than in roots. MsHSP70 was induced by heat shock, abscisic acid (ABA) and hydrogen peroxide. Transgenic Arabidopsis seedlings overexpressing MsHSP70 were hyposensitive to polyethylene glycol (PEG) and ABA treatments, suggesting that exogenous expression of MsHSP70 enhanced Arabidopsis tolerance to these stresses. Examination of physiological indexes related to drought and ABA stress demonstrated that in comparison with non-transgenic plants, T3 transgenic Arabidopsis plants had an increased proline content, higher superoxide dismutase (SOD) activity, and decreased malondialdehyde (MDA) content. Furthermore, higher relative water content (RWC) was detected in transgenic plants compared with non-transgenic plants under drought stress. These findings clearly indicate that molecular manipulation of MsHSP70 in plants can have substantial effects on stress tolerance.

  13. Expression of Helicobacter pylori hspA Gene in Lactococcus lactis NICE System and Experimental Study on Its Immunoreactivity

    Directory of Open Access Journals (Sweden)

    Xiao-Juan Zhang


    Full Text Available Aim. The aim of this study was to develop an oral Lactococcus lactis (L. lactis vaccine against Helicobacter pylori (H. pylori. Methods. After L. lactis NZ3900/pNZ8110-hspA was constructed, growth curves were plotted to study whether the growth of recombinant L. lactis was affected after hspA was cloned into L. lactis and whether the growth of empty bacteria, empty plasmid bacteria, and recombinant L. lactis was affected by different concentrations of Nisin; SDS-PAGE and Western blot were adopted, respectively, to detect the HspA expressed by recombinant L. lactis and its immunoreactivity. Results. There was no effect observed from the growth curve after exogenous gene hspA was cloned into L. lactis NZ3900; different concentrations of Nisin did not affect the growth of NZ3900 and NZ3900/pNZ8110, while different concentrations of Nisin inhibited the growth of NZ3900/pNZ8110-hspA except 10 ng/mL Nisin. No HspA strip was observed from SDS-PAGE. Western blot analysis showed that HspA expressed by recombinant bacteria had favorable immunoreactivity. Conclusion. The growth of recombinant L. lactis was suppressed even though a small amount of HspA had been induced to express. Therefore recombinant L. lactis only express HspA which was not suitable to be oral vaccine against Helicobacter pylori.

  14. Involvement of yeast HSP90 isoforms in response to stress and cell death induced by acetic acid. (United States)

    Silva, Alexandra; Sampaio-Marques, Belém; Fernandes, Angela; Carreto, Laura; Rodrigues, Fernando; Holcik, Martin; Santos, Manuel A S; Ludovico, Paula


    Acetic acid-induced apoptosis in yeast is accompanied by an impairment of the general protein synthesis machinery, yet paradoxically also by the up-regulation of the two isoforms of the heat shock protein 90 (HSP90) chaperone family, Hsc82p and Hsp82p. Herein, we show that impairment of cap-dependent translation initiation induced by acetic acid is caused by the phosphorylation and inactivation of eIF2α by Gcn2p kinase. A microarray analysis of polysome-associated mRNAs engaged in translation in acetic acid challenged cells further revealed that HSP90 mRNAs are over-represented in this polysome fraction suggesting preferential translation of HSP90 upon acetic acid treatment. The relevance of HSP90 isoform translation during programmed cell death (PCD) was unveiled using genetic and pharmacological abrogation of HSP90, which suggests opposing roles for HSP90 isoforms in cell survival and death. Hsc82p appears to promote survival and its deletion leads to necrotic cell death, while Hsp82p is a pro-death molecule involved in acetic acid-induced apoptosis. Therefore, HSP90 isoforms have distinct roles in the control of cell fate during PCD and their selective translation regulates cellular response to acetic acid stress.

  15. Organization and evolution of hsp70 clusters strikingly differ in two species of Stratiomyidae (Diptera inhabiting thermally contrasting environments

    Directory of Open Access Journals (Sweden)

    Bettencourt Brian R


    Full Text Available Abstract Background Previously, we described the heat shock response in dipteran species belonging to the family Stratiomyidae that develop in thermally and chemically contrasting habitats including highly aggressive ones. Although all species studied exhibit high constitutive levels of Hsp70 accompanied by exceptionally high thermotolerance, we also detected characteristic interspecies differences in heat shock protein (Hsp expression and survival after severe heat shock. Here, we analyzed genomic libraries from two Stratiomyidae species from thermally and chemically contrasting habitats and determined the structure and organization of their hsp70 clusters. Results Although the genomes of both species contain similar numbers of hsp70 genes, the spatial distribution of hsp70 copies differs characteristically. In a population of the eurytopic species Stratiomys singularior, which exists in thermally variable and chemically aggressive (hypersaline conditions, the hsp70 copies form a tight cluster with approximately equal intergenic distances. In contrast, in a population of the stenotopic Oxycera pardalina that dwells in a stable cold spring, we did not find hsp70 copies in tandem orientation. In this species, the distance between individual hsp70 copies in the genome is very large, if they are linked at all. In O. pardalina we detected the hsp68 gene located next to a hsp70 copy in tandem orientation. Although the hsp70 coding sequences of S. singularior are highly homogenized via conversion, the structure and general arrangement of the hsp70 clusters are highly polymorphic, including gross aberrations, various deletions in intergenic regions, and insertion of incomplete Mariner transposons in close vicinity to the 3'-UTRs. Conclusions The hsp70 gene families in S. singularior and O. pardalina evolved quite differently from one another. We demonstrated clear evidence of homogenizing gene conversion in the S. singularior hsp70 genes, which form

  16. Human heat shock protein (Hsp) 90 interferes with Neisseria meningitidis adhesin A (NadA)-mediated adhesion and invasion. (United States)

    Montanari, Paolo; Bozza, Giuseppe; Capecchi, Barbara; Caproni, Elena; Barrile, Riccardo; Norais, Nathalie; Capitani, Mirco; Sallese, Michele; Cecchini, Paola; Ciucchi, Laura; Gao, Zhenai; Rappuoli, Rino; Pizza, Mariagrazia; Aricò, Beatrice; Merola, Marcello


    NadA (N eisseria meningitidisadhesin A), a meningococcal surface protein, mediates adhesion to and invasion of human cells, an activity in which host membrane proteins have been implicated. While investigating these host factors in human epithelial cells by affinity chromatography, we discovered an unanticipated interaction of NadA with heat shock protein (Hsp) 90, a molecular chaperone. The specific in vitro interaction of recombinant soluble NadA and Hsp90 was confirmed by co-immunoprecipitations, dot and far-Western blot. Intriguingly, ADP, but not ATP, was required for this association, and the Hsp90 inhibitor 17-AAG promoted complex formation. Hsp90 binding to an Escherichia coli strain used as carrier to express surface exposed NadA confirmed these results in live bacteria. We also examined RNA interference, plasmid-driven overexpression, addition of exogenous rHsp90 and 17-AAG inhibition in human epithelial cells to further elucidate the involvement of Hsp90 in NadA-mediated adhesion and invasion. Together, these data suggest an inverse correlation between the amount of host Hsp90 and the NadA adhesive/invasive phenotype. Confocal microscopy also demonstrated that meningococci interact with cellular Hsp90, a completely novel finding. Altogether our results show that variation of host Hsp90 expression or activity interferes with adhesive and invasive events driven by NadA.

  17. Secretion of extracellular hsp90α via exosomes increases cancer cell motility: a role for plasminogen activation

    Directory of Open Access Journals (Sweden)

    Chan Doug


    Full Text Available Abstract Background Metastasis is a multi-step process that is responsible for the majority of deaths in cancer patients. Current treatments are not effective in targeting metastasis. The molecular chaperone hsp90α is secreted from invasive cancer cells and activates MMP-2 to enhance invasiveness, required for the first step in metastasis. Methods We analyzed the morphology and motility of invasive cancer cells that were treated with exogenous exosomes in the presence or absence of hsp90α. We performed mass spectrometry and immunoprecipitation to identify plasminogen as a potential client protein of extracellular hsp90α. Plasmin activation assays and migration assays were performed to test if plasminogen is activated by extracellular hsp90α and has a role in migration. Results We found that hsp90α is secreted in exosomes in invasive cancer cells and it contributes to their invasive nature. We identified a novel interaction between hsp90α and tissue plasminogen activator that together with annexin II, also found in exosomes, activates plasmin. Extracellular hsp90α promotes plasmin activation as well as increases plasmin dependent cell motility. Conclusions Our data indicate that hsp90α is released by invasive cancer cells via exosomes and implicates hsp90α in activating plasmin, a second protease that acts in cancer cell invasion.

  18. Expression of hsp70, hsp100 and ubiquitin in Aloe barbadensis Miller under direct heat stress and under temperature acclimation conditions. (United States)

    Huerta, Claudia; Freire, Matías; Cardemil, Liliana


    KEY MESSAGE : The study determined the tolerance of Aloe vera to high temperature, focusing on the expression of hsp70 , hsp100 and ubiquitin genes. These were highly expressed in plants acclimated at 35 °C prior to a heat shock of 45 °C. Aloe barbadensis Miller (Aloe vera), a CAM plant, was introduced into Chile in the semiarid IV and III Regions, which has summer diurnal temperature fluctuations of 25 to 40 °C and annual precipitation of 40 mm (dry years) to 170 mm (rainy years). The aim of this study was to investigate how Aloe vera responds to water and heat stress, focusing on the expression of heat shock genes (hsp70, hsp100) and ubiquitin, which not studied before in Aloe vera. The LT(50) of Aloe vera was determined as 53.2 °C. To study gene expression by semi-quantitative RT-PCR, primers were designed against conserved regions of these genes. Sequencing the cDNA fragments for hsp70 and ubiquitin showed a high identity, over 95 %, with the genes from cereals. The protein sequence of hsp70 deduced from the sequence of the cDNA encloses partial domains for binding ATP and the substrate. The protein sequence of ubiquitin deduced from the cDNA encloses a domain for interaction with the enzymes E2, UCH and CUE. The expression increased with temperature and water deficit. Hsp70 expression at 40-45 °C increased 50 % over the controls, while the expression increased by 150 % over the controls under a water deficit of 50 % FC. The expression of all three genes was also studied under 2 h of acclimation at 35 or 40 °C prior to a heat shock at 45 °C. Under these conditions, the plants showed greater expression of all genes than when they were subjected to direct heat stress.

  19. Influence of induced heat stress on HSP70 in buffalo lymphocytes. (United States)

    Mishra, A; Hooda, O K; Singh, G; Meur, S K


    Heat stress in farm animals, such as cattle and buffalo during summer and post-summer seasons is a problem for livestock producers. The effect of heat stress becomes pronounced when heat stress is accompanied with ambient humidity impairing the immune status, growth, production and reproductive performance of animals. Increase in HSP70 levels from cell cultures in presence of different stressors often does not reflect the physiological adaptability of animals governing thermal regulation. In this study we directly compared the effect of different heat stress conditions with the immune status and HSP70 expression patterns from buffalo lymphocytes both in vivo and in vitro. Murrah buffalo calves were exposed to induced heat stress with two experimental treatments: hot-dry (42 °C with existing relative humidity) or hot humid (35 °C with 70% relative humidity) condition in psychometric chamber, 4 h daily for 12 days and compared with control animals maintained in an experimental shed under natural conditions. There was >200-fold increase in serum-HSP70 levels in both heat stress conditions compared with control. Furthermore, the immune status of the calves failed to activate the level of HSP70 expression in serum lymphocytes. Lymphocytes cultured in vitro at higher temperature exert 2.5-fold increase in HSP70 concentration. This study is the first of its kind to demonstrate more complex expression pattern of buffalo serum-HSP70 level as a thermo adaptive response compared with in vitro treated cells. Results from this study indicate that serum-HSP70 levels could be used as a sensitive biomarker for heat stress management in large farm animals.

  20. Upon oxidative stress, the antiapoptotic Hsp60/procaspase-3 complex persists in mucoepidermoid carcinoma cells

    Directory of Open Access Journals (Sweden)

    C Campanella


    Full Text Available Hsp60, a mitochondrial chaperonin highly conserved during evolution, has been found elevated in the cytosol of cancer cells, both in vivo and in vitro, but its role in determining apoptosis during oxidative stress (OS has not yet been fully elucidated. The aim of the present work was to study the effects of OS on Hsp60 levels and its interactions with procaspase- 3 (p-C3 and p53 in tumor cells. NCI-H292 (mucoepidermoid carcinoma cells were exposed to various concentrations of hydrogen peroxide (H2O2 for 24 hours. Cell viability was determined by Trypan blue and MTT assays. DNA damage was assessed by the Comet assay, and apoptosis was measured by the AnnexinV cytofluorimetric test. Exposure to increasing concentrations of H2O2 resulted in a reduction of cell viability, DNA damage, and early apoptotic phenomena. Hsp60, p-C3, p53, and p21 were assessed by Western blotting and immunocytochemistry before and after OS. Hsp60 and p-C3 were present before and after OS induction. Immunoprecipitation experiments showed an Hsp60/p-C3 complex before OS that persisted after it, while an Hsp60/p53 complex was not detected in either condition. The presence of wild type (wt p53 was confirmed by RT-PCR, and p21 detection suggested p53 activation after OS. We postulate that, although OS may induce early apoptosis in NCI-H292 cells, Hsp60 exerts an anti-apoptotic effect in these cells and, by extension, it may do so in other cancer cells.

  1. Hsp90/p50cdc37 is required for mixed-lineage kinase (MLK) 3 signaling. (United States)

    Zhang, Hua; Wu, Wei; Du, Yan; Santos, Sarah J; Conrad, Susan E; Watson, Jack T; Grammatikakis, Nicholas; Gallo, Kathleen A


    Mixed-lineage kinase 3 (MLK3) is a mitogen-activated protein kinase (MAPK) kinase kinase that activates MAPK pathways, including the c-Jun NH(2)-terminal kinase (JNK) and p38 pathways. MLK3 and its family members have been implicated in JNK-mediated apoptosis. A survey of human cell lines revealed high levels of MLK3 in breast cancer cells. To learn more about MLK3 regulation and its signaling pathways in breast cancer cells, we engineered the estrogen-responsive human breast cancer cell line, MCF-7, to stably, inducibly express FLAG epitope-tagged MLK3. FLAG.MLK3 complexes were isolated by affinity purification, and associated proteins were identified by in-gel trypsin digestion followed by liquid chromatography/tandem mass spectrometry. Among the proteins identified were heat shock protein 90alpha,beta (Hsp90) and its kinase-specific co-chaperone p50(cdc37). We show that endogenous MLK3 complexes with Hsp90 and p50(cdc37). Further experiments demonstrate that MLK3 associates with Hsp90/p50(cdc37) through its catalytic domain in an activity-independent manner. Upon treatment of MCF-7 cells with geldanamycin, an ansamycin antibiotic that inhibits Hsp90 function, MLK3 levels decrease dramatically. Furthermore, tumor necrosis factor alpha-induced activation of MLK3 and JNK in MCF-7 cells is blocked by geldanamycin treatment. Our finding that geldanamycin treatment does not affect the cellular levels of the downstream signaling components, MAPK kinase 4, MAPK kinase 7, and JNK, suggests that Hsp90/p50(cdc37) regulates JNK signaling at the MAPK kinase kinase level. Previously identified Hsp90/p50(cdc37) clients include oncoprotein kinases and protein kinases that promote cellular proliferation and survival. Our findings reveal that Hsp90/p50(cdc37) also regulates protein kinases involved in apoptotic signaling.

  2. Short-duration-focused ultrasound stimulation of Hsp70 expression in vivo (United States)

    Kruse, D. E.; Mackanos, M. A.; O'Connell-Rodwell, C. E.; Contag, C. H.; Ferrara, K. W.


    The development of transgenic reporter mice and advances in in vivo optical imaging have created unique opportunities to assess and analyze biological responses to thermal therapy directly in living tissues. Reporter mice incorporating the regulatory regions from the genes encoding the 70 kDa heat-shock proteins (Hsp70) and firefly luciferase (luc) as reporter genes can be used to non-invasively reveal gene activation in living tissues in response to thermal stress. High-intensity-focused ultrasound (HIFU) can deliver measured doses of acoustic energy to highly localized regions of tissue at intensities that are sufficient to stimulate Hsp70 expression. We report activation of Hsp70-luc expression using 1 s duration HIFU heating to stimulate gene expression in the skin of the transgenic reporter mouse. Hsp70 expression was tracked for 96 h following the application of 1.5 MHz continuous-wave ultrasound with spatial peak intensities ranging from 53 W cm-2 up to 352 W cm-2. The results indicated that peak Hsp70 expression is observed 6-48 h post-heating, with significant activity remaining at 96 h. Exposure durations were simulated using a finite-element model, and the predicted temperatures were found to be consistent with the observed Hsp70 expression patterns. Histological evaluation revealed that the thermal damage starts at the stratum corneum and extends deeper with increasing intensity. These results indicated that short-duration HIFU may be useful for inducing heat-shock expression, and that the period between treatments needs to be greater than 96 h due to the protective properties of Hsp70.

  3. HSP33 in eukaryotes - an evolutionary tale of a chaperone adapted to photosynthetic organisms. (United States)

    Segal, Na'ama; Shapira, Michal


    HSP33 was originally identified in bacteria as a redox-sensitive chaperone that protects unfolded proteins from aggregation. Here, we describe a eukaryote ortholog of HSP33 from the green algae Chlamydomonas reinhardtii, which appears to play a protective role under light-induced oxidizing conditions. The algal HSP33 exhibits chaperone activity, as shown by citrate synthase aggregation assays. Studies from the Jakob laboratory established that activation of the bacterial HSP33 upon its oxidation initiates by the release of pre-bound Zn from the well conserved Zn-binding motif Cys-X-Cys-Xn -Cys-X-X-Cys, and is followed by significant structural changes (Reichmann et al., ). Unlike the bacterial protein, the HSP33 from C. reinhardtii had lost the first cysteine residue of its center, diminishing Zn-binding activity under all conditions. As a result, the algal protein can be easily activated by minor structural changes in response to oxidation and/or excess heat. An attempt to restore the missing first cysteine did not have a major effect on Zn-binding and on the mode of activation. Replacement of all remaining cysteines abolished completely any residual Zn binding, although the chaperone activation was maintained. A phylogenetic analysis of the algal HSP33 showed that it clusters with the cyanobacterial protein, in line with its biochemical localization to the chloroplast. Indeed, expression of the algal HSP33 increases in response to light-induced oxidative stress, which is experienced routinely by photosynthetic organisms. Despite the fact that no ortholog could be found in higher eukaryotes, its abundance in all algal species examined could have a biotechnological relevance.

  4. The molecular chaperone Hsp90 is required for cell cycle exit in Drosophila melanogaster.

    Directory of Open Access Journals (Sweden)

    Jennifer L Bandura

    Full Text Available The coordination of cell proliferation and differentiation is crucial for proper development. In particular, robust mechanisms exist to ensure that cells permanently exit the cell cycle upon terminal differentiation, and these include restraining the activities of both the E2F/DP transcription factor and Cyclin/Cdk kinases. However, the full complement of mechanisms necessary to restrain E2F/DP and Cyclin/Cdk activities in differentiating cells are not known. Here, we have performed a genetic screen in Drosophila melanogaster, designed to identify genes required for cell cycle exit. This screen utilized a PCNA-miniwhite(+ reporter that is highly E2F-responsive and results in a darker red eye color when crossed into genetic backgrounds that delay cell cycle exit. Mutation of Hsp83, the Drosophila homolog of mammalian Hsp90, results in increased E2F-dependent transcription and ectopic cell proliferation in pupal tissues at a time when neighboring wild-type cells are postmitotic. Further, these Hsp83 mutant cells have increased Cyclin/Cdk activity and accumulate proteins normally targeted for proteolysis by the anaphase-promoting complex/cyclosome (APC/C, suggesting that APC/C function is inhibited. Indeed, reducing the gene dosage of an inhibitor of Cdh1/Fzr, an activating subunit of the APC/C that is required for timely cell cycle exit, can genetically suppress the Hsp83 cell cycle exit phenotype. Based on these data, we propose that Cdh1/Fzr is a client protein of Hsp83. Our results reveal that Hsp83 plays a heretofore unappreciated role in promoting APC/C function during cell cycle exit and suggest a mechanism by which Hsp90 inhibition could promote genomic instability and carcinogenesis.

  5. The molecular chaperone Hsp90 is required for cell cycle exit in Drosophila melanogaster. (United States)

    Bandura, Jennifer L; Jiang, Huaqi; Nickerson, Derek W; Edgar, Bruce A


    The coordination of cell proliferation and differentiation is crucial for proper development. In particular, robust mechanisms exist to ensure that cells permanently exit the cell cycle upon terminal differentiation, and these include restraining the activities of both the E2F/DP transcription factor and Cyclin/Cdk kinases. However, the full complement of mechanisms necessary to restrain E2F/DP and Cyclin/Cdk activities in differentiating cells are not known. Here, we have performed a genetic screen in Drosophila melanogaster, designed to identify genes required for cell cycle exit. This screen utilized a PCNA-miniwhite(+) reporter that is highly E2F-responsive and results in a darker red eye color when crossed into genetic backgrounds that delay cell cycle exit. Mutation of Hsp83, the Drosophila homolog of mammalian Hsp90, results in increased E2F-dependent transcription and ectopic cell proliferation in pupal tissues at a time when neighboring wild-type cells are postmitotic. Further, these Hsp83 mutant cells have increased Cyclin/Cdk activity and accumulate proteins normally targeted for proteolysis by the anaphase-promoting complex/cyclosome (APC/C), suggesting that APC/C function is inhibited. Indeed, reducing the gene dosage of an inhibitor of Cdh1/Fzr, an activating subunit of the APC/C that is required for timely cell cycle exit, can genetically suppress the Hsp83 cell cycle exit phenotype. Based on these data, we propose that Cdh1/Fzr is a client protein of Hsp83. Our results reveal that Hsp83 plays a heretofore unappreciated role in promoting APC/C function during cell cycle exit and suggest a mechanism by which Hsp90 inhibition could promote genomic instability and carcinogenesis.

  6. Single nucleotide polymorphisms in HSP17.8 and their association with agronomic traits in barley.

    Directory of Open Access Journals (Sweden)

    Yanshi Xia

    Full Text Available Small heat shock protein 17.8 (HSP17.8 is produced abundantly in plant cells under heat and other stress conditions and may play an important role in plant tolerance to stress environments. However, HSP17.8 may be differentially expressed in different accessions of a crop species exposed to identical stress conditions. The ability of different genotypes to adapt to various stress conditions resides in their genetic diversity. Allelic variations are the most common forms of genetic variation in natural populations. In this study, single nucleotide polymorphisms (SNPs of the HSP17.8 gene were investigated across 210 barley accessions collected from 30 countries using EcoTILLING technology. Eleven SNPs including 10 from the coding region of HSP17.8 were detected, which form nine distinguishable haplotypes in the barley collection. Among the 10 SNPs in the coding region, six are missense mutations and four are synonymous nucleotide changes. Five of the six missense changes are predicted to be deleterious to HSP17.8 function. The accessions from Middle East Asia showed the higher nucleotide diversity of HSP17.8 than those from other regions and wild barley (H. spontaneum accessions exhibited greater diversity than the cultivated barley (H. vulgare accessions. Four SNPs in HSP17.8 were found associated with at least one of the agronomic traits evaluated except for spike length, namely number of grains per spike, thousand kernel weight, plant height, flag leaf area and leaf color. The association between SNP and these agronomic traits may provide new insight for study of the gene's potential contribution to drought tolerance of barley.

  7. Single nucleotide polymorphisms in HSP17.8 and their association with agronomic traits in barley. (United States)

    Xia, Yanshi; Li, Ronghua; Ning, Zhengxiang; Bai, Guihua; Siddique, Kadambot H M; Yan, Guijun; Baum, Michael; Varshney, Rajeev K; Guo, Peiguo


    Small heat shock protein 17.8 (HSP17.8) is produced abundantly in plant cells under heat and other stress conditions and may play an important role in plant tolerance to stress environments. However, HSP17.8 may be differentially expressed in different accessions of a crop species exposed to identical stress conditions. The ability of different genotypes to adapt to various stress conditions resides in their genetic diversity. Allelic variations are the most common forms of genetic variation in natural populations. In this study, single nucleotide polymorphisms (SNPs) of the HSP17.8 gene were investigated across 210 barley accessions collected from 30 countries using EcoTILLING technology. Eleven SNPs including 10 from the coding region of HSP17.8 were detected, which form nine distinguishable haplotypes in the barley collection. Among the 10 SNPs in the coding region, six are missense mutations and four are synonymous nucleotide changes. Five of the six missense changes are predicted to be deleterious to HSP17.8 function. The accessions from Middle East Asia showed the higher nucleotide diversity of HSP17.8 than those from other regions and wild barley (H. spontaneum) accessions exhibited greater diversity than the cultivated barley (H. vulgare) accessions. Four SNPs in HSP17.8 were found associated with at least one of the agronomic traits evaluated except for spike length, namely number of grains per spike, thousand kernel weight, plant height, flag leaf area and leaf color. The association between SNP and these agronomic traits may provide new insight for study of the gene's potential contribution to drought tolerance of barley.

  8. Administration of M. leprae Hsp65 interferes with the murine lupus progression.

    Directory of Open Access Journals (Sweden)

    Eliana B Marengo

    Full Text Available The heat shock protein [Hsp] family guides several steps during protein synthesis, are abundant in prokaryotic and eukaryotic cells, and are highly conserved during evolution. The Hsp60 family is involved in assembly and transport of proteins, and is expressed at very high levels during autoimmunity or autoinflammatory phenomena. Here, the pathophysiological role of the wild type [WT] and the point mutated K(409A recombinant Hsp65 of M. leprae in an animal model of Systemic Lupus Erythematosus [SLE] was evaluated in vivo using the genetically homogeneous [NZBxNZW]F(1 mice. Anti-DNA and anti-Hsp65 antibodies responsiveness was individually measured during the animal's life span, and the mean survival time [MST] was determined. The treatment with WT abbreviates the MST in 46%, when compared to non-treated mice [p<0.001]. An increase in the IgG2a/IgG1 anti-DNA antibodies ratio was also observed in animals injected with the WT Hsp65. Incubation of BALB/c macrophages with F(1 serum from WT treated mice resulted in acute cell necrosis; treatment of these cells with serum from K(409A treated mice did not cause any toxic effect. Moreover, the involvement of WT correlates with age and is dose-dependent. Our data suggest that Hsp65 may be a central molecule intervening in the progression of the SLE, and that the point mutated K(409A recombinant immunogenic molecule, that counteracts the deleterious effect of WT, may act mitigating and delaying the development of SLE in treated mice. This study gives new insights into the general biological role of Hsp and the significant impact of environmental factors during the pathogenesis of this autoimmune process.

  9. One out of four: HspL but no other small heat shock protein of Agrobacterium tumefaciens acts as efficient virulence-promoting VirB8 chaperone.

    Directory of Open Access Journals (Sweden)

    Yun-Long Tsai

    Full Text Available Alpha-crystallin-type small heat shock proteins (sHsps are ubiquitously distributed in most eukaryotes and prokaryotes. Four sHsp genes named hspL, hspC, hspAT1, and hspAT2 were identified in Agrobacterium tumefaciens, a plant pathogenic bacterium capable of unique interkingdom DNA transfer via type IV secretion system (T4SS. HspL is highly expressed in virulence-induced growth condition and functions as a VirB8 chaperone to promote T4SS-mediated DNA transfer. Here, we used genetic and biochemical approaches to investigate the involvement of the other three sHsps in T4SS and discovered the molecular basis underlying the dominant function of HspL in promoting T4SS function. While single deletion of hspL but no other sHsp gene reduced T4SS-mediated DNA transfer and tumorigenesis efficiency, additional deletion of other sHsp genes in the hspL deletion background caused synergistic effects in the virulence phenotypes. This is correlated with the high induction of hspL and only modest increase of hspC, hspAT1, and hspAT2 at their mRNA and protein abundance in virulence-induced growth condition. Interestingly, overexpression of any single sHsp gene alone in the quadruple mutant caused increased T4SS-mediated DNA transfer and tumorigenesis. Thermal aggregation protecting assays in vitro indicated that all four sHsps exhibit chaperone activity for the model substrate citrate synthase but only HspL functions as efficient chaperone for VirB8. The higher VirB8 chaperone activity of HspL was also demonstrated in vivo, in which lower amounts of HspL than other sHsps were sufficient in maintaining VirB8 homeostasis in A. tumefaciens. Domain swapping between HspL and HspAT2 indicated that N-terminal, central alpha-crystallin, and C-terminal domains of HspL all contribute to HspL function as an efficient VirB8 chaperone. Taken together, we suggest that the dominant role of HspL in promoting T4SS function is based on its higher expression in virulence

  10. Bio-batteries and bio-fuel cells: leveraging on electronic charge transfer proteins. (United States)

    Kannan, A M; Renugopalakrishnan, V; Filipek, S; Li, P; Audette, G F; Munukutla, L


    Bio-fuel cells are alternative energy devises based on bio-electrocatalysis of natural substrates by enzymes or microorganisms. Here we review bio-fuel cells and bio-batteries based on the recent literature. In general, the bio-fuel cells are classified based on the type of electron transfer; mediated electron transfer and direct electron transfer or electronic charge transfer (ECT). The ECT of the bio-fuel cells is critically reviewed and a variety of possible applications are considered. The technical challenges of the bio-fuel cells, like bioelectrocatalysis, immobilization of bioelectrocatalysts, protein denaturation etc. are highlighted and future research directions are discussed leveraging on the use of electron charge transfer proteins. In addition, the packaging aspects of the bio-fuel cells are also analyzed and the found that relatively little work has been done in the engineering development of bio-fuel cells.

  11. Types for BioAmbients

    Directory of Open Access Journals (Sweden)

    Sara Capecchi


    Full Text Available The BioAmbients calculus is a process algebra suitable for representing compartmentalization, molecular localization and movements between compartments. In this paper we enrich this calculus with a static type system classifying each ambient with group types specifying the kind of compartments in which the ambient can stay. The type system ensures that, in a well-typed process, ambients cannot be nested in a way that violates the type hierarchy. Exploiting the information given by the group types, we also extend the operational semantics of BioAmbients with rules signalling errors that may derive from undesired ambients' moves (i.e. merging incompatible tissues. Thus, the signal of errors can help the modeller to detect and locate unwanted situations that may arise in a biological system, and give practical hints on how to avoid the undesired behaviour.

  12. Types for BioAmbients

    CERN Document Server

    Capecchi, Sara; 10.4204/EPTCS.19.7


    The BioAmbients calculus is a process algebra suitable for representing compartmentalization, molecular localization and movements between compartments. In this paper we enrich this calculus with a static type system classifying each ambient with group types specifying the kind of compartments in which the ambient can stay. The type system ensures that, in a well-typed process, ambients cannot be nested in a way that violates the type hierarchy. Exploiting the information given by the group types, we also extend the operational semantics of BioAmbients with rules signalling errors that may derive from undesired ambients' moves (i.e. merging incompatible tissues). Thus, the signal of errors can help the modeller to detect and locate unwanted situations that may arise in a biological system, and give practical hints on how to avoid the undesired behaviour.

  13. NMR and Mutational Identification of the Collagen-Binding Site of the Chaperone Hsp47 (United States)

    Yagi-Utsumi, Maho; Yoshikawa, Sumi; Yamaguchi, Yoshiki; Nishi, Yohei; Kurimoto, Eiji; Ishida, Yoshihito; Homma, Takayuki; Hoseki, Jun; Nishikawa, Yoshimi; Koide, Takaki; Nagata, Kazuhiro; Kato, Koichi


    Heat shock protein 47 (Hsp47) acts as a client-specific chaperone for collagen and plays a vital role in collagen maturation and the consequent embryonic development. In addition, this protein can be a potential target for the treatment of fibrosis. Despite its physiological and pathological importance, little is currently known about the collagen-binding mode of Hsp47 from a structural aspect. Here, we describe an NMR study that was conducted to identify the collagen-binding site of Hsp47. We used chicken Hsp47, which has higher solubility than its human counterpart, and applied a selective 15N-labeling method targeting its tryptophan and histidine residues. Spectral assignments were made based on site-directed mutagenesis of the individual residues. By inspecting the spectral changes that were observed upon interaction with a trimeric collagen peptide and the mutational data, we successfully mapped the collagen-binding site in the B/C β-barrel domain and a nearby loop in a 3D-homology model based upon a serpin fold. This conclusion was confirmed by mutational analysis. Our findings provide a molecular basis for the design of compounds that target the interaction between Hsp47 and procollagen as therapeutics for fibrotic diseases. PMID:23049894

  14. NMR and mutational identification of the collagen-binding site of the chaperone Hsp47.

    Directory of Open Access Journals (Sweden)

    Maho Yagi-Utsumi

    Full Text Available Heat shock protein 47 (Hsp47 acts as a client-specific chaperone for collagen and plays a vital role in collagen maturation and the consequent embryonic development. In addition, this protein can be a potential target for the treatment of fibrosis. Despite its physiological and pathological importance, little is currently known about the collagen-binding mode of Hsp47 from a structural aspect. Here, we describe an NMR study that was conducted to identify the collagen-binding site of Hsp47. We used chicken Hsp47, which has higher solubility than its human counterpart, and applied a selective (15N-labeling method targeting its tryptophan and histidine residues. Spectral assignments were made based on site-directed mutagenesis of the individual residues. By inspecting the spectral changes that were observed upon interaction with a trimeric collagen peptide and the mutational data, we successfully mapped the collagen-binding site in the B/C β-barrel domain and a nearby loop in a 3D-homology model based upon a serpin fold. This conclusion was confirmed by mutational analysis. Our findings provide a molecular basis for the design of compounds that target the interaction between Hsp47 and procollagen as therapeutics for fibrotic diseases.

  15. Sequence Analysis of HSP70 Gene of Leishmania major and Leishmania tropica in Chabahar and Mashhad

    Directory of Open Access Journals (Sweden)

    Mansour Dabirzadeh


    Full Text Available Background and Objective: Cutaneous leishmaniasis is a parasitic disease and a health problem in different parts of Iran, especially two cities of Mashhad and Chabahar. Due to morphological similarities of most Leishmania species and difference in reservoirs of L. major and L. tropica, it is necessary to determine the parasite specie to combat the disease. Thus, this study used gene sequencing and genotyping of 70-kDa heat shock protein (HSP70 to differentiate the two species of Leishmania. Methods: In this descriptive-analytical study, microscope slides and cultures were prepared from 43 patients suspected of cutaneous leishmaniasis in Chabahar and Mashhad. PCR was performed after genomic DNA extraction and then PCR products were sequenced and analyzed. Results: Of the 43 patients studied, 32 direct smear and culture (74.4% were positive and 11 (25.6% showed negative results, and were therefore excluded from the study. Using HSP70-specific primers, 1962 bp and 1152bp bands were observed for HSP70 of L. major in Chabahar and L. tropica in Mashhad, respectively. Based on the results, there were 18 nucleotide differences between HSP70 of L. major in Chabahar and L. tropica in Mashhad. Conclusion: Due to the morphological similarities between Leishmania species and inability to differentiate species through parasitological methods, the HSP70 gene can be used for identification of the species, and prevention and treatment of the disease.

  16. Changes of levels of hs-CRP, Hsp70, and MPO in patients with preeclampsia

    Institute of Scientific and Technical Information of China (English)

    Xia Liu


    Objective:To observe the changes of the levels of hs-CRP, Hsp70, and MPO in patients with preeclampsia (PE) and their clinical significance. Methods:A total of 60 pregnant women with mild and severe PE who were admitted in our hospital from January, 2013 to March, 2014 were included in the study and divided into the mild group and the severe group with 30 cases in each group. The levels of serum hs-CRP, Hsp70, and MPO were detected, and their relations with PE were analyzed. The lipid metabolism related indicators were meanwhile detected. A total of 30 healthy later pregnant women at the same stage were served as the control group for comparative analysis. Results:The levels of serum hs-CRP, Hsp70, and MPO in the mild and severe PE group were significantly higher than those in the control group (P0.05). Conclusions: The abnormal lipid metabolism in different degrees exists in the pregnant women with PE, and hs-CRP, Hsp70, and MPO are together involved in the pathogenesis of PE. Detection of the levels of hs-CRP, Hsp70, and MPO in the pregnant women with PE can accurately reflect the condition of PE.

  17. PTEX component HSP101 mediates export of diverse malaria effectors into host erythrocytes. (United States)

    Beck, Josh R; Muralidharan, Vasant; Oksman, Anna; Goldberg, Daniel E


    To mediate its survival and virulence, the malaria parasite Plasmodium falciparum exports hundreds of proteins into the host erythrocyte. To enter the host cell, exported proteins must cross the parasitophorous vacuolar membrane (PVM) within which the parasite resides, but the mechanism remains unclear. A putative Plasmodium translocon of exported proteins (PTEX) has been suggested to be involved for at least one class of exported proteins; however, direct functional evidence for this has been elusive. Here we show that export across the PVM requires heat shock protein 101 (HSP101), a ClpB-like AAA+ ATPase component of PTEX. Using a chaperone auto-inhibition strategy, we achieved rapid, reversible ablation of HSP101 function, resulting in a nearly complete block in export with substrates accumulating in the vacuole in both asexual and sexual parasites. Surprisingly, this block extended to all classes of exported proteins, revealing HSP101-dependent translocation across the PVM as a convergent step in the multi-pathway export process. Under export-blocked conditions, association between HSP101 and other components of the PTEX complex was lost, indicating that the integrity of the complex is required for efficient protein export. Our results demonstrate an essential and universal role for HSP101 in protein export and provide strong evidence for PTEX function in protein translocation into the host cell.

  18. HSP60, a protein downregulated by IGFBP7 in colorectal carcinoma

    Directory of Open Access Journals (Sweden)

    Lin Jie


    Full Text Available Abstract Background In our previous study, it was well defined that IGFBP7 was an important tumor suppressor gene in colorectal cancer (CRC. We aimed to uncover the downstream molecules responsible for IGFBP7's behaviour in this study. Methods Differentially expressed protein profiles between PcDNA3.1(IGFBP7-transfected RKO cells and the empty vector transfected controls were generated by two-dimensional gel electrophoresis (2-DE and mass spectrometry (MS identification. The selected differentially expressed protein induced by IGFBP7 was confirmed by western blot and ELISA. The biological behaviour of the protein was explored by cell growth assay and colony formation assay. Results Six unique proteins were found differentially expressed in PcDNA3.1(IGFBP7-transfected RKO cells, including albumin (ALB, 60 kDa heat shock protein(HSP60, Actin cytoplasmic 1 or 2, pyruvate kinase muscle 2(PKM2, beta subunit of phenylalanyl-tRNA synthetase(FARSB and hypothetical protein. The downregulation of HSP60 by IGFBP7 was confirmed by western blot and ELISA. Recombinant human HSP60 protein could increase the proliferation rate and the colony formation ability of PcDNA3.1(IGFBP7-RKO cells. Conclusion HSP60 was an important downstream molecule of IGFBP7. The downregulation of HSP60 induced by IGFBP7 may be, at least in part, responsible for IGFBP7's tumor suppressive biological behaviour in CRC.

  19. Defining Hsp70 Subnetworks in Dengue Virus Replication Reveals Key Vulnerability in Flavivirus Infection. (United States)

    Taguwa, Shuhei; Maringer, Kevin; Li, Xiaokai; Bernal-Rubio, Dabeiba; Rauch, Jennifer N; Gestwicki, Jason E; Andino, Raul; Fernandez-Sesma, Ana; Frydman, Judith


    Viral protein homeostasis depends entirely on the machinery of the infected cell. Accordingly, viruses can illuminate the interplay between cellular proteostasis components and their distinct substrates. Here, we define how the Hsp70 chaperone network mediates the dengue virus life cycle. Cytosolic Hsp70 isoforms are required at distinct steps of the viral cycle, including entry, RNA replication, and virion biogenesis. Hsp70 function at each step is specified by nine distinct DNAJ cofactors. Of these, DnaJB11 relocalizes to virus-induced replication complexes to promote RNA synthesis, while DnaJB6 associates with capsid protein and facilitates virion biogenesis. Importantly, an allosteric Hsp70 inhibitor, JG40, potently blocks infection of different dengue serotypes in human primary blood cells without eliciting viral resistance or exerting toxicity to the host cells. JG40 also blocks replication of other medically-important flaviviruses including yellow fever, West Nile and Japanese encephalitis viruses. Thus, targeting host Hsp70 subnetworks provides a path for broad-spectrum antivirals.

  20. Impact of mild temperature hardening on thermotolerance, fecundity, and Hsp gene expression in Liriomyza huidobrensis. (United States)

    Huang, Li-Hua; Chen, Bing; Kang, Le


    The pea leafminer, Liriomyza huidobrensis, is one of the most important economic insect pests around the world. Its population fluctuates greatly with seasonal change in China, and temperature was thought to be one of the important reasons. In attempt to further explore the impact of disadvantageous temperature on L. huidobrensis, 1-day-old adults were shocked at various temperatures (10, 25, 32, and 35 degrees C, respectively) for 4h, and the effects on thermotolerance, feeding, and fecundity were studied. Meanwhile the expression of five heat shock genes (hsp90, 70, 60, 40, and 20) was examined by real-time quantitative PCR. Our results showed that both 32 and 35 degrees C hardenings remarkably increased adult heat resistance, whereas cold tolerance was not improved accordingly. No cross resistance in response to cold and heat stresses was observed. Both adult feeding and fecundity were dramatically reduced, but no effect was observed on egg hatching, larval survival, pupal eclosion, or sex ratio. The results indicate that the deleterious effect on fecundity is the result of direct cessation of oviposition during the period of stress. Simultaneously, the mRNA levels of hsp70 and hsp20 significantly increased upon thermal hardening. Taken together, our results suggest that mild heat hardening improves thermotolerance of L. huidobrensis at the cost of impairment on fecundity, and the induced expression of hsp70 and hsp20 may play an important role in balancing the functional tradeoff.

  1. Hsp90 as a Gatekeeper of Tumor Angiogenesis: Clinical Promise and Potential Pitfalls

    Directory of Open Access Journals (Sweden)

    J. E. Bohonowych


    Full Text Available Tumor vascularization is an essential modulator of early tumor growth, progression, and therapeutic outcome. Although antiangiogenic treatments appear promising, intrinsic and acquired tumor resistance contributes to treatment failure. Clinical inhibition of the molecular chaperone heat shock protein 90 (Hsp90 provides an opportunity to target multiple aspects of this signaling resiliency, which may elicit more robust and enduring tumor repression relative to effects elicited by specifically targeted agents. This review highlights several primary effectors of angiogenesis modulated by Hsp90 and describes the clinical challenges posed by the redundant circuitry of these pathways. The four main topics addressed include (1 Hsp90-mediated regulation of HIF/VEGF signaling, (2 chaperone-dependent regulation of HIF-independent VEGF-mediated angiogenesis, (3 Hsp90-dependent targeting of key proangiogenic receptor tyrosine kinases and modulation of drug resistance, and (4 consideration of factors such as tumor microenvironment that pose several challenges for the clinical efficacy of anti-angiogenic therapy and Hsp90-targeted strategies.

  2. Designed hybrid TPR peptide targeting Hsp90 as a novel anticancer agent

    Directory of Open Access Journals (Sweden)

    Ohara Koji


    Full Text Available Abstract Background Despite an ever-improving understanding of the molecular biology of cancer, the treatment of most cancers has not changed dramatically in the past three decades and drugs that do not discriminate between tumor cells and normal tissues remain the mainstays of anticancer therapy. Since Hsp90 is typically involved in cell proliferation and survival, this is thought to play a key role in cancer, and Hsp90 has attracted considerable interest in recent years as a potential therapeutic target. Methods We focused on the interaction of Hsp90 with its cofactor protein p60/Hop, and engineered a cell-permeable peptidomimetic, termed "hybrid Antp-TPR peptide", modeled on the binding interface between the molecular chaperone Hsp90 and the TPR2A domain of Hop. Results It was demonstrated that this designed hybrid Antp-TPR peptide inhibited the interaction of Hsp90 with the TPR2A domain, inducing cell death of breast, pancreatic, renal, lung, prostate, and gastric cancer cell lines in vitro. In contrast, Antp-TPR peptide did not affect the viability of normal cells. Moreover, analysis in vivo revealed that Antp-TPR peptide displayed a significant antitumor activity in a xenograft model of human pancreatic cancer in mice. Conclusion These results indicate that Antp-TPR peptide would provide a potent and selective anticancer therapy to cancer patients.

  3. Heat Shock Protein (HSP) Drug Discovery and Development: Targeting Heat Shock Proteins in Disease (United States)

    Shrestha, Liza; Bolaender, Alexander; Patel, Hardik J.; Taldone, Tony


    Heat shock proteins (HSPs) present as a double edged sword. While they play an important role in maintaining protein homeostasis in a normal cell, cancer cells have evolved to co-opt HSP function to promote their own survival. As a result, HSPs such as HSP90 have attracted a great deal of interest as a potential anticancer target. These efforts have resulted in over 20 distinct compounds entering clinical evaluation for the treatment of cancer. However, despite the potent anticancer activity demonstrated in preclinical models, to date no HSP90 inhibitor has obtained regulatory approval. In this review we discuss the unique challenges faced in targeting HSPs that have likely contributed to their lack of progress in the clinic and suggest ways to overcome these so that the enormous potential of these compounds to benefit patients can finally be realized. We also provide a guideline for the future development of HSP-targeted agents based on the many lessons learned during the last two decades in developing HSP90 inhibitors. PMID:27072696

  4. Hsp90 in the continuum of breast ductal carcinogenesis: Evaluation in precursors, preinvasive and ductal carcinoma lesions

    Directory of Open Access Journals (Sweden)

    Patsouris Effstratios


    Full Text Available Abstract Background Hsp90 (heat shock protein90 is a chaperone protein essential for preserving and regulating the function of various cellular proteins. Elevated Hsp90 expression seems to be a trait of breast cancer and may be an integral part of the coping mechanisms that cancer cells exhibit vis-à-vis stress. This manuscript tries to examine the immunohistochemical expression of Hsp90 all along the continuum of breast ductal lesions encompassing ductal hyperplasia without atypia (DHWithoutA, atypical ductal hyperplasia (ADH, ductal carcinoma in situ (DCIS and invasive ductal carcinoma (IDC. Methods Tissue specimens were taken from 30 patients with DHWithoutA, 31 patients with ADH, 51 with DCIS and 51 with IDC. Immunohistochemical assessment of Hsp90 was performed both in the lesion and the adjacent normal breast ducts and lobules; the latter serving as control. Concerning Hsp90 assessment the percentage of positive cells and the intensity were separately analyzed. Subsequently, the Allred score was calculated. Post hoc analysis on the correlations between Hsp90 Allred score and possible predictors (grade, nodal status, tumor size, ER Allred score, PR Allred score, c-erbB-2 status and triple negative status was conducted in IDC. Results Hsp90 exhibited mainly cytoplasmic immunoreactivity. Hsp90 Allred score exhibited an increasing trend along the continuum of breast ductal lesions (Spearman's rho = 0.169, p = 0.031. Compared to the adjacent normal ducts and lobules, no statistically significant differences were noted in DHwithoutA, ADH and DCIS. Hsp90 expression (intensity, positive cells, Allred score was higher in IDC, compared to the adjacent normal tissue. Higher Hsp90 expression was observed in grade 2/3 IDCs (borderline association and tumors of larger size. At the univariable analysis, higher Hsp90 expression was associated with higher ER Allred score, PR Allred score and c-erbB-2 positivity in IDC. Triple-negative IDCs exhibited

  5. Plasmodium falciparum Hep1 Is Required to Prevent the Self Aggregation of PfHsp70-3.

    Directory of Open Access Journals (Sweden)

    David O Nyakundi

    Full Text Available The majority of mitochondrial proteins are encoded in the nucleus and need to be imported from the cytosol into the mitochondria, and molecular chaperones play a key role in the efficient translocation and proper folding of these proteins in the matrix. One such molecular chaperone is the eukaryotic mitochondrial heat shock protein 70 (Hsp70; however, it is prone to self-aggregation and requires the presence of an essential zinc-finger protein, Hsp70-escort protein 1 (Hep1, to maintain its structure and function. PfHsp70-3, the only Hsp70 predicted to localize in the mitochondria of P. falciparum, may also rely on a Hep1 orthologue to prevent self-aggregation. In this study, we identified a putative Hep1 orthologue in P. falciparum and co-expression of PfHsp70-3 and PfHep1 enhanced the solubility of PfHsp70-3. PfHep1 suppressed the thermally induced aggregation of PfHsp70-3 but not the aggregation of malate dehydrogenase or citrate synthase, thus showing specificity for PfHsp70-3. Zinc ions were indeed essential for maintaining the function of PfHep1, as EDTA chelation abrogated its abilities to suppress the aggregation of PfHsp70-3. Soluble and functional PfHsp70-3, acquired by co-expression with PfHep-1, will facilitate the biochemical characterisation of this particular Hsp70 protein and its evaluation as a drug target for the treatment of malaria.

  6. Significant correlation between expression level of HSP gp96 and progression of hepatitis B virus induced diseases

    Institute of Scientific and Technical Information of China (English)

    Xiao-Dong Zhu; Cheng-Lin Li; Zhen-Wei Lang; George F Gao; Po Tien


    AIM: Gp96, also known as Grp94, is a member of heat shock protein (HSP) family and binds repertoires of peptides thereof eliciting peptide-specific T cell immune responses.It predominantly locates inside the endoplasmic reticulum (ER) with some cell surface expression in certain cancerous cells. Previous studies have shown that gp96 expression level was up-regulated in tumor cells, including hepatocellular carcinoma (HCC). However, relationship between the extent of gp96 expression and disease progression especially HBV-induced chronic infection, cirrhosis and hepatocellular carcinoma, has not been addressed before. As primary HCC can be induced and progressed from chronic hepatitis B virus (HBV) infection and HBV-induced cirrhosis, we designed an immunohistochemical experiment to test the correlation between gp96 expression level and HBV-induced disease progression, from chronic HBV infection, cirrhosis to HCC.METHODS: We chose liver samples from different patients of hepatitis B virus induced diseases, including chronic hepatitis B (77 patients), cirrhosis (27 patients) and primary HCC (30 patients), to test the expression level of gp96 in different affected groups. Formalin-fixed, and paraffinembedded liver tissues taken from these patients were immuno-stained by using an anti-gp96 monoclonal antibody for the expression level of gp96 protein in the sections. In addition, Western blotting of whole cell lysates derived from established human embryonic liver cell lines and several human HCC cell lines (Huh7, HepG2, SSMC-7721) was compared with the expression of gp96.RESULTS: We found that the extent of elevated gp96expression was significantly correlated with the disease progression, and was the highest in HCC patients, lowest in chronic HBV infection and was that of the cirrhosis in the middle.CONCLUSION: Increased expression of gp96 might be used as a diagnostic or prognostic bio-marker for the HBV infection and HBV-induced diseases.

  7. The stress kit: A new method based on competitive reverse transcriptase-polymerase chain reaction to quantify the expression of human αB-crystallin, Hsp27, and Hsp60

    NARCIS (Netherlands)

    Bajramović, J.J.; Geutskens, S.B.; Bsibsi, M.; Boot, M.; Hassankhan, R.; Verhulst, K.C.; Noort, J.M. van


    We describe a reverse transcriptase-polymerase chain reaction method for the semiquantitative detection of mRNAs encoding the human heat shock proteins αβ-crystallin, Hsp27, and Hsp60. The method involves the coamplification of cellular mRNA-derived cDNA with a dilution series of a competitor fragme

  8. Cloning and Expression of HSP of Taenia Solium Oncosphere%猪带绦虫六钩蚴 HSP 的克隆和表达

    Institute of Scientific and Technical Information of China (English)

    王哲; 赵权


      The HSP gene was separately amplified from total RNA of activated Taenia solium oncosphere by RT -PCR.The PCR products were cloned into pGH vector,recombinant positive clones was sequenced after restriction enzyme digestion.The HSP gene was subcloned into pET28a expression vector,the recombinant pET28a-HSP infected into E.coliBL21.IPTG was added to induce fusion expression and the expression products was identified by SDS-PAGE and Western-blot.One fusion protein band about 35 kDa was identified by SDS -PAGE after inducible expression after inducible expression,The result would lay foundations for the mechanism of invasion of between oncosphere and host,and the design of new vaccine anti-porcine cysticercosis and taeniasis.%  提取猪带绦虫激活和未激活六钩蚴总 RNA,RT-PCR 扩增 HSP 目的基因,将目的基因与 pGH 克隆载体连接,经酶切鉴定后,将阳性重组质粒进行测序,结果扩增出激活的六钩蚴的目的片段。将目的基因亚克隆到原核表达载体 pET-28a-HSP中,并将获得的 pET28a-HSP 阳性重组子转化至宿主菌 E.coliBL21,IPTG 进行诱导表达,并对重组抗原 pET-28a-HSP 进行SDS-PAGE 和 Western-blot 检测。结果表明重组经 SDS-PAGE 分析可见一条约35 kDa 大小的融合蛋白条带的抗原,Western-blot 结果显示其能被囊虫病人阳性血清识别。这将为进一步阐明六钩蚴入侵中间宿主的机理、设计新型抗猪囊虫病和绦虫病疫苗打下基础。

  9. Molecular docking study, synthesis and biological evaluation of Schiff bases as Hsp90 inhibitors. (United States)

    Dutta Gupta, Sayan; Snigdha, D; Mazaira, Gisela I; Galigniana, Mario D; Subrahmanyam, C V S; Gowrishankar, N L; Raghavendra, N M


    Heat shock protein 90 (Hsp90) is an emerging attractive target for the discovery of novel cancer therapeutic agents. Docking methods are powerful in silico tools for lead generation and optimization. In our mission to rationally develop novel effective small molecules against Hsp90, we predicted the potency of our designed compounds by Sybyl surflex Geom X docking method. The results of the above studies revealed that Schiff bases derived from 2,4-dihydroxy benzaldehyde/5-chloro-2,4-dihydroxy benzaldehyde demonstrated effective binding with the protein. Subsequently, a few of them were synthesized (1-10) and characterized by IR, (1)HNMR and mass spectral analysis. The synthesized molecules were evaluated for their potential to suppress Hsp90 ATPase activity by Malachite green assay. The anticancer studies were performed by 3-(4,5-dimethythiazol- 2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay method. The software generated results was in satisfactory agreement with the evaluated biological activity.

  10. Functional analysis of Drosophila HSP70 promoter with different HSE numbers in human cells. (United States)

    Kust, Nadezda; Rybalkina, Ekaterina; Mertsalov, Ilya; Savchenko, Ekaterina; Revishchin, Alexander; Pavlova, Gali


    The activation of genetic constructs including the Drosophila hsp70 promoter with four and eight HSE sequences in the regulatory region has been described in human cells. The promoter was shown to be induced at lower temperatures compared to the human hsp70 promoter. The promoter activity increased after a 60-min heat shock already at 38 °C in human cells. The promoter activation was observed 24 h after heat shock for the constructs with eight HSEs, while those with four HSEs required 48 h. After transplantation of in vitro heat-shocked transfected cells, the promoter activity could be maintained for 3 days with a gradual decline. The promoter activation was confirmed in vivo without preliminary heat shock in mouse ischemic brain foci. Controlled expression of the Gdnf gene under a Drosophila hsp70 promoter was demonstrated. This promoter with four and eight HSE sequences in the regulatory region can be proposed as a regulated promoter in genetic therapeutic systems.

  11. Enhancement of Hsp70 synthesis protects common carp, Cyprinus carpio L., against lethal ammonia toxicity. (United States)

    Sung, Y Y; Roberts, R J; Bossier, P


    Exposure to TEX-OE®, a patented extract of the prickly pear cactus (Opuntia ficus indica) containing chaperone-stimulating factor, was shown to protect common carp, Cyprinus carpio L., fingerlings against acute ammonia stress. Survival was enhanced twofold from 50% to 95% after exposure to 5.92 mg L(-1) NH(3) , a level determined in the ammonia challenge bioassay as the 1-h LD50 concentration for this species. Survival of TEX-OE®-pre-exposed fish was enhanced by 20% over non-exposed controls during lethal ammonia challenge (14.21 mg L(-1)  NH(3) ). Increase in the levels of gill and muscle Hsp70 was evident in TEX-OE®-pre-exposed fish but not in the unexposed controls, indicating that application of TEX-OE® accelerated carp endogenous Hsp70 synthesis during ammonia perturbation. Protection against ammonia was correlated with Hsp70 accretion.

  12. Exploring in vitro and in vivo Hsp90 inhibitors activity against human protozoan parasites. (United States)

    Giannini, Giuseppe; Battistuzzi, Gianfranco


    A set of compounds, previously selected as potent Hsp90α inhibitors, has been studied on a panel of human parasites. 5-Aryl-3,4-isoxazolediamide derivatives (1) were active against two protozoa, Trypanosoma brucei rhodesiense and Plasmodium falciparum, with a good tolerability toward cytotoxicity on non-malignant L6 rat myoblast cell line, unlike the 1,5-diaryl,4-carboxamides-1,2,3-triazole derivatives (2) which, while showing a single-digit nM range activity against the same protozoa, were also highly cytotoxic on L6 cells. In a subsequent in vivo study, two isoxazolediamide derivatives, 1a and 1b, were very efficacious on the sleeping sickness-causing agent with a clear parasitaemia during treatment. These data, however, showed that not all protozoa are sensitive to Hsp90 inhibitors, as well as not all Hsp90 inhibitors are equally active on parasites.

  13. Spontaneous assembly of HSP90 inhibitors at water/octanol interface: A molecular dynamics simulation study (United States)

    Zolghadr, Amin Reza; Boroomand, Samaneh


    Drug absorption at an acceptable dose depends on the pair of solubility and permeability. There are many potent therapeutics that are not active in vivo, presumably due to the lack of capability to cross the cell membrane. Molecular dynamics simulation of radicicol, diol-radicicol, cyclopropane-radicicol and 17-DMAG were performed at water/octanol interface to suggest interfacial activity as a physico-chemical characteristic of these heat shock protein 90 (HSP90) inhibitors. We have observed that orally active HSP90 inhibitors form aggregates at the water/octanol and DPPC-lipid/water interfaces by starting from an initial configuration with HSP90 inhibitors embedded in the water matrix.

  14. Real-time observation of the conformational dynamics of mitochondrial Hsp70 by spFRET. (United States)

    Sikor, Martin; Mapa, Koyeli; von Voithenberg, Lena Voith; Mokranjac, Dejana; Lamb, Don C


    The numerous functions of the important class of molecular chaperones, heat shock proteins 70 (Hsp70), rely on cycles of intricate conformational changes driven by ATP-hydrolysis and regulated by cochaperones and substrates. Here, we used Förster resonance energy transfer to study the conformational dynamics of individual molecules of Ssc1, a mitochondrial Hsp70, in real time. The intrinsic dynamics of the substrate-binding domain of Ssc1 was observed to be uncoupled from the dynamic interactions between substrate- and nucleotide-binding domains. Analysis of the fluctuations in the interdomain separation revealed frequent transitions to a nucleotide-free state. The nucleotide-exchange factor Mge1 did not induce ADP release, as expected, but rather facilitated binding of ATP. These results indicate that the conformational cycle of Ssc1 is more elaborate than previously thought and provide insight into how the Hsp70s can perform a wide variety of functions.

  15. Bioética Bioética Boethics


    Cássia Regina Rodrigues Nunes; Amauri Porto Nunes


    A bioética é um novo campo de conhecimento e uma corrente de pensamento que vem se desenvolvendo amplamente no Brasil, principalmente a partir da década de 90. Nesse artigo buscamos contextualizar o seu surgimento, justificando a necessidade da criação desse neologismo, uma vez que a ética estava de certo modo esquecida, em relação à técnica, restringindo-se apenas à ética profissional. Hoje nos deparamos com uma ampla gama de produções nessa área que ajudam a nortear nossa ação enquanto cida...

  16. Bioética Bioética Boethics

    Directory of Open Access Journals (Sweden)

    Cássia Regina Rodrigues Nunes


    Full Text Available A bioética é um novo campo de conhecimento e uma corrente de pensamento que vem se desenvolvendo amplamente no Brasil, principalmente a partir da década de 90. Nesse artigo buscamos contextualizar o seu surgimento, justificando a necessidade da criação desse neologismo, uma vez que a ética estava de certo modo esquecida, em relação à técnica, restringindo-se apenas à ética profissional. Hoje nos deparamos com uma ampla gama de produções nessa área que ajudam a nortear nossa ação enquanto cidadãos que buscam qualidade de vida e a preservação da vida no planeta. A bioética, no entanto, preocupa-se com o modo de ser: pensamento-julgamento-ação, em relação aos seres humanos entre si e com a natureza.La bioética es un nuevo campo de conocimiento y una corriente del pensamiento que está en pleno desarrollo en Brasil, principalmente a partir de la década del 90. En este artículo explicamos el contexto de su surgimiento, justificando la necesidad de la creación de este neologismo, una vez que la ética estaba de una cierta manera relegada, con relación a la técnica, limitada apenas a la ética profesional. Hoy nosotros notamos una amplia gama de producciones en esta área que nos ayudan a orientar nuestras acciones como ciudadanos que buscan la calidad de vida y la preservación de la vida en el planeta. La bioética, sin embargo, se preocupa con el modo de ser: pensamiento-juicio-acción, con relación a los seres humanos entre sí y con la naturaleza.Bioethics is a new field of knowledge and a current of thought that has been largely developing in Brazil, especially since the Nineties. In this article, we try to contextualize its inception by justifying the need to create such a neologism, since ethics was somewhat forgotten, in relation to the technique, being restricted to professional ethics. We are nowadays faced with a wide scope of works in this area that help guide our action as citizens who want quality of life

  17. Hsp10 nuclear localization and changes in lung cells response to cigarette smoke suggest novel roles for this chaperonin. (United States)

    Corrao, Simona; Anzalone, Rita; Lo Iacono, Melania; Corsello, Tiziana; Di Stefano, Antonino; D'Anna, Silvestro Ennio; Balbi, Bruno; Carone, Mauro; Sala, Anna; Corona, Davide; Timperio, Anna Maria; Zolla, Lello; Farina, Felicia; de Macario, Everly Conway; Macario, Alberto J L; Cappello, Francesco; La Rocca, Giampiero


    Heat-shock protein (Hsp)10 is the co-chaperone for Hsp60 inside mitochondria, but it also resides outside the organelle. Variations in its levels and intracellular distribution have been documented in pathological conditions, e.g. cancer and chronic obstructive pulmonary disease (COPD). Here, we show that Hsp10 in COPD undergoes changes at the molecular and subcellular levels in bronchial cells from human specimens and derived cell lines, intact or subjected to stress induced by cigarette smoke extract (CSE). Noteworthy findings are: (i) Hsp10 occurred in nuclei of epithelial and lamina propria cells of bronchial mucosa from non-smokers and smokers; (ii) human bronchial epithelial (16HBE) and lung fibroblast (HFL-1) cells, in vitro, showed Hsp10 in the nucleus, before and after CSE exposure; (iii) CSE stimulation did not increase the levels of Hsp10 but did elicit qualitative changes as indicated by molecular weight and isoelectric point shifts; and (iv) Hsp10 nuclear levels increased after CSE stimulation in HFL-1, indicating cytosol to nucleus migration, and although Hsp10 did not bind DNA, it bound a DNA-associated protein.

  18. [The role of heat shock proteins HSP90 in the response of immune cells to centimeter microwaves]. (United States)

    Glushkova, O V; Novoselova, E G; Khrenov, M O; Novoselova, T V; Cherenkov, D A; Lunin, S M; Fesenko, E E


    The effects of low-level electromagnetic waves (8.15-18 GHz, 1 microW/cm2, 1 h) on the production of heat shock proteins, several cytokines, and nitric oxide in isolated mouse macrophages and lymphocytes were examined both under normal conditions and after the treatment of the cells with geldanamycin (GA), a depressor of activity of the heat shock protein 90 (Hsp90). The irradiation of cells without GA induced the production of Hsp70, nitric oxide (NO), interleukin-1beta (IL-1beta), interleukin-10 (IL-10), and the tumor necrosis factor -alpha (TNF-alpha). No changes in the production of Hsp90 in irradiated cells were observed, but intracellular locations of Hsp25 and Hsp70 altered. The preliminary treatment of cells with GA did not remove the effects of microwaves: in these conditions, the synthesis of all cytokines tested, nitric oxide, as well as total and membrane amount of Hsp70, and the amount of Hsp25 in the cytoplasm and cytoskeleton increased. Moreover, the exposure of cells incubated with GA resulted in the reduction of Hsp90-alpha production.

  19. Hsp10 nuclear localization and changes in lung cells response to cigarette smoke suggest novel roles for this chaperonin (United States)

    Corrao, Simona; Anzalone, Rita; Lo Iacono, Melania; Corsello, Tiziana; Di Stefano, Antonino; D'Anna, Silvestro Ennio; Balbi, Bruno; Carone, Mauro; Sala, Anna; Corona, Davide; Timperio, Anna Maria; Zolla, Lello; Farina, Felicia; Conway de Macario, Everly; Macario, Alberto J. L.; Cappello, Francesco; La Rocca, Giampiero


    Heat-shock protein (Hsp)10 is the co-chaperone for Hsp60 inside mitochondria, but it also resides outside the organelle. Variations in its levels and intracellular distribution have been documented in pathological conditions, e.g. cancer and chronic obstructive pulmonary disease (COPD). Here, we show that Hsp10 in COPD undergoes changes at the molecular and subcellular levels in bronchial cells from human specimens and derived cell lines, intact or subjected to stress induced by cigarette smoke extract (CSE). Noteworthy findings are: (i) Hsp10 occurred in nuclei of epithelial and lamina propria cells of bronchial mucosa from non-smokers and smokers; (ii) human bronchial epithelial (16HBE) and lung fibroblast (HFL-1) cells, in vitro, showed Hsp10 in the nucleus, before and after CSE exposure; (iii) CSE stimulation did not increase the levels of Hsp10 but did elicit qualitative changes as indicated by molecular weight and isoelectric point shifts; and (iv) Hsp10 nuclear levels increased after CSE stimulation in HFL-1, indicating cytosol to nucleus migration, and although Hsp10 did not bind DNA, it bound a DNA-associated protein. PMID:25355063

  20. Characterization of a wheat HSP70 gene and its expression in response to stripe rust infection and abiotic stresses

    NARCIS (Netherlands)

    Duan, Y.H.; Guo, J.; Ding, K.; Wang, S.J.; Zhang, H.; Dai, X.W.; Chen, Y.Y.; Govers, F.; Huang, L.L.; Kang, Z.S.


    Members of the family of 70-kD heat shock proteins (HSP70 s) play various stress-protective roles in plants. In this study, a wheat HSP70 gene was isolated from a suppression subtractive hybridization (SSH) cDNA library of wheat leaves infected by Puccinia striiformis f. sp. tritici. The gene, that

  1. Role of heat shock protein Hsp25 in the response of the orofacial nuclei motor system to physiological stress (United States)

    Murashov, A. K.; Talebian, S.; Wolgemuth, D. J.


    Although expression of the small heat shock protein family member Hsp25 has been previously observed in the central nervous system (CNS), both constitutively and upon induction, its function in the CNS remains far from clear. In the present study we have characterized the spatial pattern of expression of Hsp25 in the normal adult mouse brain as well as the changes in expression patterns induced by subjecting mice to experimental hyperthermia or hypoxia. Immunohistochemical analysis revealed a surprisingly restricted pattern of constitutive expression of Hsp25 in the brain, limited to the facial, trigeminal, ambiguus, hypoglossal and vagal motor nuclei of the brainstem. After hyperthermia or hypoxia treatment, significant increases in the levels of Hsp25 were observed in these same areas and also in fibers of the facial and trigeminal nerve tracts. Immunoblot analysis of protein lysates from brainstem also showed the same pattern of induction of Hsp25. Surprisingly, no other area in the brain showed expression of Hsp25, in either control or stressed animals. The highly restricted expression of Hsp25 implies that this protein may have a specific physiological role in the orofacial motor nuclei, which govern precise coordination between muscles of mastication and the pharynx, larynx, and face. Its rapid induction after stress further suggests that Hsp25 may serve as a specific molecular chaperone in the lower cholinergic motor neurons and along their fibers under conditions of stress or injury. Copyright 1998 Elsevier Science B.V.

  2. Involvement of serum HSP 70 in Guillain-Barré Syndrome: An exploratory study and a review of current literature

    Directory of Open Access Journals (Sweden)

    Aida Loshaj-Shala


    Full Text Available The evolutionary conserved family of heat shock proteins (HSP is responsible for protecting cells against different types of stress. Although the levels of HSP can be readily measured in serum, the levels of HSP 70 in patients Guillain-Barre Syndrome (GBS have not been studied before. To this aim we investigate whether patients with GBS (n=21 had altered serum HSP 70 levels compared to healthy controls (HC, n=9 and to patients affected by other immune disorders such as multifocal motor neuropathy (MMN, n=4 and chronic inflammatory demyelinating polyneuropathy (CIDP, n=6. The highest HSP 70 value (15.78 ± 1.72 ng/mL was found in one patient in the GBS group, although we have found that serum HSP70 levels were significantly higher in 2 out of the 21 GBS patients (9.5%. Hence, it is of interest to underline that the patient with the highest HSP70 level, had also the best recovery rate. Моrе extensive research is required in order to support the hypothesis that HSP 70 serum concentration may be a useful biomarker for the prediction of remission outcome for GBS patients.

  3. Influenza A virus nucleoprotein exploits Hsp40 to inhibit PKR activation.

    Directory of Open Access Journals (Sweden)

    Kulbhushan Sharma

    Full Text Available BACKGROUND: Double-stranded RNA dependent protein kinase (PKR is a key regulator of the anti-viral innate immune response in mammalian cells. PKR activity is regulated by a 58 kilo Dalton cellular inhibitor (P58(IPK, which is present in inactive state as a complex with Hsp40 under normal conditions. In case of influenza A virus (IAV infection, P58(IPK is known to dissociate from Hsp40 and inhibit PKR activation. However the influenza virus component responsible for PKR inhibition through P58(IPK activation was hitherto unknown. PRINCIPAL FINDINGS: Human heat shock 40 protein (Hsp40 was identified as an interacting partner of Influenza A virus nucleoprotein (IAV NP using a yeast two-hybrid screen. This interaction was confirmed by co-immunoprecipitation studies from mammalian cells transfected with IAV NP expressing plasmid. Further, the IAV NP-Hsp40 interaction was validated in mammalian cells infected with various seasonal and pandemic strains of influenza viruses. Cellular localization studies showed that NP and Hsp40 co-localize primarily in the nucleus. During IAV infection in mammalian cells, expression of NP coincided with the dissociation of P58(IPK from Hsp40 and decrease PKR phosphorylation. We observed that, plasmid based expression of NP in mammalian cells leads to decrease in PKR phosphorylation. Furthermore, inhibition of NP expression during influenza virus replication led to PKR activation and concomitant increase in eIF2α phosphorylation. Inhibition of NP expression also led to reduced IRF3 phosphorylation, enhanced IFN β production and concomitant reduction of virus replication. Taken together our data suggest that NP is the viral factor responsible for P58(IPK activation and subsequent inhibition of PKR-mediated host response during IAV infection. SIGNIFICANCE: Our findings demonstrate a novel role of IAV NP in inhibiting PKR-mediated anti-viral host response and help us understand P58(IPK mediated inhibition of PKR activity

  4. Antigen-presenting cells transfected with Hsp65 messenger RNA fail to treat experimental tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, C.D.; Trombone, A.P.F.; Lorenzi, J.C.C.; Almeida, L.P.; Gembre, A.F.; Padilha, E. [Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Ramos, S.G. [Departamento de Patologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil); Silva, C.L.; Coelho-Castelo, A.A.M. [Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP (Brazil)


    In the last several years, the use of dendritic cells has been studied as a therapeutic strategy against tumors. Dendritic cells can be pulsed with peptides or full-length protein, or they can be transfected with DNA or RNA. However, comparative studies suggest that transfecting dendritic cells with messenger RNA (mRNA) is superior to other antigen-loading techniques in generating immunocompetent dendritic cells. In the present study, we evaluated a new therapeutic strategy to fight tuberculosis using dendritic cells and macrophages transfected with Hsp65 mRNA. First, we demonstrated that antigen-presenting cells transfected with Hsp65 mRNA exhibit a higher level of expression of co-stimulatory molecules, suggesting that Hsp65 mRNA has immunostimulatory properties. We also demonstrated that spleen cells obtained from animals immunized with mock and Hsp65 mRNA-transfected dendritic cells were able to generate a mixed Th1/Th2 response with production not only of IFN-γ but also of IL-5 and IL-10. In contrast, cells recovered from mice immunized with Hsp65 mRNA-transfected macrophages were able to produce only IL-5. When mice were infected with Mycobacterium tuberculosis and treated with antigen-presenting cells transfected with Hsp65 mRNA (therapeutic immunization), we did not detect any decrease in the lung bacterial load or any preservation of the lung parenchyma, indicating the inability of transfected cells to confer curative effects against tuberculosis. In spite of the lack of therapeutic efficacy, this study reports for the first time the use of antigen-presenting cells transfected with mRNA in experimental tuberculosis.

  5. Heritability of hsp70 expression in the beetle Tenebrio molitor: Ontogenetic and environmental effects. (United States)

    Lardies, Marco A; Arias, María Belén; Poupin, María Josefina; Bacigalupe, Leonardo D


    Ectotherms constitute the vast majority of terrestrial biodiversity and are especially likely to be vulnerable to climate warming because their basic physiological functions such as locomotion, growth, and reproduction are strongly influenced by environmental temperature. An integrated view about the effects of global warming will be reached not just establishing how the increase in mean temperature impacts the natural populations but also establishing the effects of the increase in temperature variance. One of the molecular responses that are activated in a cell under a temperature stress is the heat shock protein response (HSP). Some studies that have detected consistent differences among thermal treatments and ontogenetic stages in HSP70 expression have assumed that these differences had a genetic basis and consequently expression would be heritable. We tested for changes in quantitative genetic parameters of HSP70 expression in a half-sib design where individuals of the beetle Tenebrio molitor were maintained in constant and varying thermal environments. We estimated heritability of HSP70 expression using a linear mixed modelling approach in different ontogenetic stages. Expression levels of HSP70 were consistently higher in the variable environment and heritability estimates were low to moderate. The results imply that within each ontogenetic stage additive genetic variance was higher in the variable environment and in adults compared with constant environment and larvae stage, respectively. We found that almost all the genetic correlations across ontogenetic stages and environment were positive. These suggest that directional selection for higher levels of expression in one environment will result in higher expression levels of HSP70 on the other environment for the same ontogenetic stage.

  6. The Bio* toolkits--a brief overview. (United States)

    Mangalam, Harry


    Bioinformatics research is often difficult to do with commercial software. The Open Source BioPerl, BioPython and Biojava projects provide toolkits with multiple functionality that make it easier to create customised pipelines or analysis. This review briefly compares the quirks of the underlying languages and the functionality, documentation, utility and relative advantages of the Bio counterparts, particularly from the point of view of the beginning biologist programmer.

  7. Biodistribution profiles of recombinant anti-breast cancer immunotherapeutic infusion protein HSP-MUC1 vaccine after subcutaneous in tumor-bearing mice

    Institute of Scientific and Technical Information of China (English)

    JingBAI; Hai-fengSONG; Xiu-wenLIU; Bao-zhenZHU; LunOU; XiaoSUN; Xiao-junMIAO


    AIM: To study biodistributional profiles of recombinant antibreast cancer immunotherapeutic fusion protein HSP-MUC1 vaccine after subcutaneous in tumor-bearing mice. METHODS: HSP-MUC1 of variant tissue was detected by 125I-HSP-MUC1 combined with trichloroacetic acid (TCA) precipitation or sizeexclusive high performance liquid chromatography (SHPLC). RESULTS: 125I-HSP-MUC1 distributed widely. The highest

  8. Study on relationship between porcine HSP70 protein expression and weaning diarrhoea piglets%仔猪HSP70蛋白表达与断奶腹泻关系研究

    Institute of Scientific and Technical Information of China (English)

    王莉; 汪蕊; 王希彪


    为了探讨HSP70在断奶过程中的作用,试验通过免疫组化方法检测民猪、长白猪断奶仔猪的胃、十二指肠、空肠、回肠、盲肠、结肠六个组织中HSP70蛋白的组织定位与定量变化.结果表明,胃肠道中HSP70蛋白主要在黏膜上皮和肠腺内表达,在各组织内的表达没有因腹泻与否而发生组织定位的改变,在除胃以外的其他消化道组织中,长白健康组仔猪的HSP70蛋白的表达量显著高于长白腹泻组、民猪健康与腹泻组.%In order to investigate the effect of HSP70 during weaning of piglets, this experiment determined the expression of HSP70 protein and located HSP70 protein in the gastrointestinal (Gl) tissues (stomach, duodenum, jejunum, ileum, cecum and colon) of Min pig and Landrace by immunohistochemistry. The results showed that, HSP70 protein expressed mainly in the epithelium mucosae and intestinal gland of Gl tract, while the diarrhoea didn't affect HSP70's location. The HSP70 expression of Landrace healthy piglets in Gl tract, expect for the stomach, was significantly higher than other groups (Landrace diarrhoea piglets, Min pig healthy and diarrhoea piglets).

  9. Bio-logic设备优点

    Institute of Scientific and Technical Information of China (English)


    @@ 世界第一台数字脑电图是Bio-logic厂家设计并生产 世界第一台数字视频脑电图是Bio-logic厂家设计并生产 在中国市场"癫痫刀"这个概念,也为Bio-logic厂家所推广 在美国市场,脑电图仪器的行业标准是以Bio-logic产品为基准定制的.

  10. Bio-Matematik ved Roskilde Universitet

    DEFF Research Database (Denmark)

    Ottesen, Johnny T.


    Ved Institut for Natur, Systemer og Modeller ved Roskilde Universitet finder man en velkonsolideret gruppe af forskere, BioMath-gruppen, der beskæftiger sig med Bio-Matematik og Bio-Medicinsk Modellering. Faktisk startede BioMath-gruppen på det tidligere Institut for Studiet af Matematik og Fysik...... pumper; 3) Endokrin fysiologi; 4) Type 1 og type 2 diabetes; 5) Populationsdynamik og sygdomsspredning; og 6) Parameterestimering i patient-specifikke modeller.   Udover overnævnte modelleringsområder har vi ved instituttet en stærk tradition for at beskæftige os med metaaspekter af modellering, f...

  11. Authentifizierung von Bio-Milch im Labor



    In Deutschland ist die Nachfrage nach Bio-Lebensmitteln in den letzten Jahren stetig gestiegen. So erhöhte sich der Absatz von Bio-Trinkmilch in 2007 im Vergleich zum Vorjahr erneut kräftig um 34 Prozent (ZMP, Bonn) und der Bio-Anteil beträgt inzwischen bei Frischmilch knapp elf Prozent. Aufgrund sporadisch resultierender Lieferengpässe bei Bio-Milch sowie der vorhandenen Handelspreisdifferenz besteht zunehmend ein potenzielles Risiko der Falschdeklaration konventionell erzeugter Milch als Bi...

  12. Large-Scale Conformational Transitions and Dimerization Are Encoded in the Amino-Acid Sequences of Hsp70 Chaperones (United States)

    Malinverni, Duccio; Marsili, Simone; Barducci, Alessandro; De Los Rios, Paolo


    Hsp70s are a class of ubiquitous and highly conserved molecular chaperones playing a central role in the regulation of proteostasis in the cell. Hsp70s assist a myriad of cellular processes by binding unfolded or misfolded substrates during a complex biochemical cycle involving large-scale structural rearrangements. Here we show that an analysis of coevolution at the residue level fully captures the characteristic large-scale conformational transitions of this protein family, and predicts an evolutionary conserved–and thus functional–homo-dimeric arrangement. Furthermore, we highlight that the features encoding the Hsp70 dimer are more conserved in bacterial than in eukaryotic sequences, suggesting that the known Hsp70/Hsp110 hetero-dimer is a eukaryotic specialization built on a pre-existing template. PMID:26046683

  13. Experimental and Structural Testing Module to Analyze Paralogue-Specificity and Affinity in the Hsp90 Inhibitors Series (United States)

    Taldone, Tony; Patel, Pallav D.; Patel, Maulik; Patel, Hardik J.; Evans, Christopher E.; Rodina, Anna; Ochiana, Stefan; Shah, Smit K.; Uddin, Mohammad; Gewirth, Daniel; Chiosis, Gabriela


    We here describe the first reported comprehensive analysis of Hsp90 paralogue affinity and selectivity in the clinical Hsp90 inhibitor chemotypes. This has been possible through the development of a versatile experimental assay based on a new FP-probe (16a) that we both describe here. The assay can test rapidly and accurately the binding affinity of all major Hsp90 chemotypes and has a testing range that spans low nanomolar to millimolar binding affinities. We couple this assay with a computational analysis that allows for rationalization of paralogue selectivity and defines not only the major binding modes that relay pan-paralogue binding or, conversely, paralogue selectivity, but also identifies molecular characteristics that impart such features. The methods developed here provide a blueprint for parsing out the contribution of the four Hsp90 paralogues to the perceived biological activity with the current Hsp90 chemotypes and set the ground for the development of paralogue selective inhibitors. PMID:23965125

  14. Polymorphisms of hsp60 gene in Chinese Han people%hsp60基因多态性在中国汉族人中的分布

    Institute of Scientific and Technical Information of China (English)

    王静; 杨晓波; 白云; 江琦; 何越峰; 陈永文; 何美安; 邬堂春


    目的 检测中国汉族人热休克蛋白60(heat shock protein 60,hsp60)基因调控区和编码区的单核苷酸多态性(single nucleotide polymorphisms,SNPs),寻找hsp60基因的遗传标记.方法 采用分片段扩增直接测序的方法检测60名汉族人hsp60基因的5'侧翼区、部分编码区和内含子区,将测序结果与美国国家生物技术信息中心(National Center for Biotechnology Information,NCBI)及国际人类基因组单体型图计划(International HapMap Project,HapMap)中日本人、欧洲人及非洲人的序列进行对比,确定中国汉族人群hsp60基因SNPs的位置、类型和频率.结果 在hsp60基因2号外显子区发现1个新的同义SNP,位于2号染色体198189061位,最小等位基因频率(the minor allele frequency,MAF)为0.025.检测区域内包含11个已报道的SNPs,7个未在研究人群中检出;4个被检出的SNPs为rs1116734、rs3749095、rs1050347、rs8539,MAF分别为0.51、0.30、0.29、0.49;rs1116734、rs1050347、rs8539在日本人和中国汉族人群中的分布相似;rs3749095在中国汉族人中较常见,在日本人种中未检出;rs8539在欧洲及非洲人中的分布与在中国人的分布差异有统计学意义(P<0.01).结论 中国汉族人群hsp60的SNPs分布有别于其他人种,rs1116734、rs3749095、rs1050347、rs8539是hsp60基因在中国汉族人群中较常见的SNPs,可为Hsp60蛋白功能研究及hsp60基因与疾病关联研究候选SNPs的选择提供科学依据.

  15. Profitable use of bio fuels

    Energy Technology Data Exchange (ETDEWEB)

    Ekelund, Mats [Strateco Develoment AB, Vega (Sweden)], e-mail:


    Traditionally, the transportation industry has been opposed to any new legislation and when rather stringent emission legislation occurred, they objected just as they did when new fuels came on the agenda. On very short notice, Taxi Stockholm lost 20 % of their business when the County decided to award all public transportation contracts to a competitor. It was time to change plans instead of complaining and to take advantage of new opportunities - 'The first mover advantage'. Making the use of bio fuels into a profitable business takes a change of a standard 'business model' to do and there is still much room others to do the same. With a new CEO, an active marketing department and active individuals among the Board of Directors, Taxi Stockholm massaged a strategy where more business and private customers would be attracted by justifying the green leaf on every cab. All initiatives were publically announced and Taxi Stockholm broke new ice by putting a ban on spike tires - a decision which the vice Mayor made part of her ruling for the whole city. The Ban on gasoline and diesel cars were announced and such a statement attracted business from a loyalty point of view and from companies that had a 'Green Transport Policy' to live up to. Taxi Stockholm has seen growth and profitability grow since and credit the green policy on bio fuels such as bio gas and ethanol for most of it. Preem, Stockholm Transit, Volvo and other market driven operators have all seen markets grow from green initiatives.

  16. Study of the role of Hsp70.1 in oridonin-treated hepatocarcinoma cells%Hsp70.1在冬凌草甲素处理的肝癌细胞中的作用研究

    Institute of Scientific and Technical Information of China (English)

    王辉; 叶燕; 黄树林; 禹志领


    目的 研究Hsp70.1在冬凌草甲素处理后的肝癌细胞HepG2中的表达情况及其与冬凌草甲素抗肿瘤活性的关系.方法 采用蛋白印迹法及实时荧光定量PCR技术观察Hsp70.1在冬凌草甲素处理的肝癌细胞HepG2的表达变化,并采用RNA干扰技术观察抑制Hsp70.1表达后对冬凌草甲素抗肝癌效果的影响.结果 冬凌草甲素作用12 h后,HepG2细胞的Hsp70.1蛋白及RNA表达均增加.Hsp70.1短发夹RNA(shRNA)转染抑制Hsp70.1表达后,与对照组相比,Hsp70.1 shRNA转染组、冬凌草甲素处理组或两者联合应用组的HepG2细胞的存活率均明显降低.结论 Hsp70.1可促进HepG2细胞的生存,对HepG2细胞和冬凌草甲素处理的HepG2细胞具有保护作用.%Objective To explore the expression of Hsp70. 1 in oridonin-treated hepatocarcinoma HepC2 cells and the correlation of Hsp70. 1 with the anticancer activity of oridonin. Methods By using Western blot and real-time PCR analysis,we tested the expression of Hsp70. 1 protein and mRNA in HepG2 cells treated with oridonin. After HepG2 cells were transfected with Hsp70.1 shRNA.the influence of Hsp70.1 on the anticancer activity of oridonin was investigated. Results The levels of Hsp70. 1 protein and mRNA increased in oridonin-treated HepG2 cells. Transient transfection of Hsp70. 1-specific shRNA suppressed Hsp70. 1 expression. Compared with control cells,Hsp70.1 shRNA-transfection,oridonin-treatment and combination of both-treatment groups significantly reduced cell viability. Conclusion Hsp70. 1 upregulation contributes to HepG2 cells survival,which suggests that Hsp70. 1 acts as a protective factor in both HepG2 cells and oridonin-treated HepG2 cells.

  17. Hsp90 is important for fecundity, longevity, and buffering of cryptic deleterious variation in wild fly populations

    Directory of Open Access Journals (Sweden)

    Chen Bing


    Full Text Available Abstract Background In the laboratory, the Drosophila melanogaster heat shock protein Hsp90 can buffer the phenotypic effects of genetic variation. Laboratory experiments either manipulate Hsp90 activity pharmacologically, or they induce mutations with strong effects in the gene Hsp83, the single-copy fly gene encoding Hsp90. It is unknown whether observations from such laboratory experiments are relevant in the wild. Results We here study naturally occurring mutations in Hsp83, and their effects on fitness and phenotypic buffering in flies derived from wild populations. We examined more than 4500 flies from 42 Drosophila populations distributed world-wide for insertions or deletions of mobile DNA in or near the Hsp83 gene. The insertions we observed occur at low population frequencies, and reduce Hsp83 gene expression. In competition experiments, mutant flies performed much more poorly than wild-type flies. Mutant flies were also significantly less fecund and shorter-lived than wild-type flies, as well as less well buffered against cryptic deleterious variation, as we show through inbreeding experiments. Specifically, in Hsp83 mutant flies female fecundity dropped to much lower levels after inbreeding than in wild-type flies. At even slightly elevated temperatures, inbred mutant Hsp83 populations went extinct, whereas inbred wild-type populations persisted. Conclusions Our work shows that Hsp90, a regulator of the stress response and of signaling, helps buffer deleterious variation in fruit flies derived from wild population, and that its buffering role becomes even more important under heat stress.

  18. Evaluation of immune effect of recombinant fusion protein targeting the prostate stem cell antigen based on PSCA and HSP70

    Directory of Open Access Journals (Sweden)

    Lei DONG


    Full Text Available Objective To explore the immune effect and antitumor activity of recombinant prostate stem cell protein (PSCA and heat shock protein 70 (HSP70 in a murine model of prostate cancer. Methods Twenty-five healthy male C57BL/6 mice were randomly divided into 5 groups (5 each: PSCA, HSP, PSCA+HSP, PSCA-HSP and control group. Mice in the first 4 groups were vaccinated with the corresponding proteins, and those in control group were faked with injection of phosphate buffer saline (PBS. After immunization with recombinant proteins, the PSCA-specific cellular immune responses were monitored with ELISPOT, intracellular cytokine staining assay, and flow cytometry, and ELISA assay was used to detect humoral immune responses. The tumor growth and survival of vaccined mice were observed. Results ELISPOT revealed that the mice in PSCA-HSP group generated much more IFN-γ spot-forming cells than those in other groups (P<0.05, and they could generate strong anti-PSCA antibody response. Results of flow cytometry showed that the number of CD8+/IFN-γ+ T cells was significantly higher in PSCAHSP group than that in other groups (P<0.05. ELISA results revealed that all the mice in PSCA, PSCA+HSP and PSCA-HSP group were induced to generate the PSCA-specific humoral immune response, and no statistical difference was found on the antibody levels among the three groups. Animal experiment showed that PSCA-HSP could inhibit the growth of PSCA-expressing tumors and prolong the survival time of vaccinated mice. Conclusion HSP70 is a chaperone with significant effect for protein vaccines, and the recombinant fusion protein PSCA-HSP70 could be of potential value for prostate cancer treatment. DOI: 10.11855/j.issn.0577-7402.2014.09.08

  19. Structural and biochemical studies on Vibrio cholerae Hsp31 reveals a novel dimeric form and Glutathione-independent Glyoxalase activity (United States)

    Dey, Sanjay


    Vibrio cholerae experiences a highly hostile environment at human intestine which triggers the induction of various heat shock genes. The hchA gene product of V. cholerae O395, referred to a hypothetical intracellular protease/amidase VcHsp31, is one such stress-inducible homodimeric protein. Our current study demonstrates that VcHsp31 is endowed with molecular chaperone, amidopeptidase and robust methylglyoxalase activities. Through site directed mutagenesis coupled with biochemical assays on VcHsp31, we have confirmed the role of residues in the vicinity of the active site towards amidopeptidase and methylglyoxalase activities. VcHsp31 suppresses the aggregation of insulin in vitro in a dose dependent manner. Through crystal structures of VcHsp31 and its mutants, grown at various temperatures, we demonstrate that VcHsp31 acquires two (Type-I and Type-II) dimeric forms. Type-I dimer is similar to EcHsp31 where two VcHsp31 monomers associate in eclipsed manner through several intersubunit hydrogen bonds involving their P-domains. Type-II dimer is a novel dimeric organization, where some of the intersubunit hydrogen bonds are abrogated and each monomer swings out in the opposite directions centering at their P-domains, like twisting of wet cloth. Normal mode analysis (NMA) of Type-I dimer shows similar movement of the individual monomers. Upon swinging, a dimeric surface of ~400Å2, mostly hydrophobic in nature, is uncovered which might bind partially unfolded protein substrates. We propose that, in solution, VcHsp31 remains as an equilibrium mixture of both the dimers. With increase in temperature, transformation to Type-II form having more exposed hydrophobic surface, occurs progressively accounting for the temperature dependent increase of chaperone activity of VcHsp31. PMID:28235098

  20. LPS-induced delayed preconditioning is mediated by Hsp90 and involves the heat shock response in mouse kidney.

    Directory of Open Access Journals (Sweden)

    Tamás Kaucsár

    Full Text Available We and others demonstrated previously that preconditioning with endotoxin (LPS protected from a subsequent lethal LPS challenge or from renal ischemia-reperfusion injury (IRI. LPS is effective in evoking the heat shock response, an ancient and essential cellular defense mechanism, which plays a role in resistance to, and recovery from diseases. Here, by using the pharmacological Hsp90 inhibitor novobiocin (NB, we investigated the role of Hsp90 and the heat shock response in LPS-induced delayed renal preconditioning.Male C57BL/6 mice were treated with preconditioning (P: 2 mg/kg, i.p. and subsequent lethal (L: 10 mg/kg, i.p. doses of LPS alone or in combination with NB (100 mg/kg, i.p.. Controls received saline (C or NB.Preconditioning LPS conferred protection from a subsequent lethal LPS treatment. Importantly, the protective effect of LPS preconditioning was completely abolished by a concomitant treatment with NB. LPS induced a marked heat shock protein increase as demonstrated by Western blots of Hsp70 and Hsp90. NB alone also stimulated Hsp70 and Hsp90 mRNA but not protein expression. However, Hsp70 and Hsp90 protein induction in LPS-treated mice was abolished by a concomitant NB treatment, demonstrating a NB-induced impairment of the heat shock response to LPS preconditioning.LPS-induced heat shock protein induction and tolerance to a subsequent lethal LPS treatment was prevented by the Hsp90 inhibitor, novobiocin. Our findings demonstrate a critical role of Hsp90 in LPS signaling, and a potential involvement of the heat shock response in LPS-induced preconditioning.