A comparative genomic analysis of the small heat shock proteins in Caenorhabditis elegans and briggsae

The small heat shock proteins (sHSPs) are a ubiquitous family of molecular chaperones. We have identified 18 sHSPs in the Caenorhabditis elegans genome and 20 sHSPs in the Caenorhabditis briggsae genome. Analysis of phylogenetic relationships and evolutionary dynamics of the sHSPs in these two genomes reveals a very complex pattern of evolution. The sHSPs in C. elegans and C. briggsae do not display clear orthologous relationships with other invertebrate sHSPs. But many sHSPs in C. elegans have orthologs in C. briggsae. One group of sHSPs, the HSP16s, has a very unusual evolutionary history. Although there are a number of HSP16s in both the C. elegans and C. briggsae genomes, none of the HSP16s display orthologous relationships across these two species. The HSP16s have an unusual gene pair structure and a complex evolutionary history shaped by gene duplication, gene conversion, and purifying selection. We found no evidence of recent positive selection acting on any of the sHSPs in C. elegans or in C. briggsae. There is also no evidence of functional divergence within the pairs of orthologous C. elegans and C. briggsae sHSPs. However, the evolutionary patterns do suggest that functional divergence has occurred between the sHSPs in C. elegans and C. briggsae and the sHSPs in more distantly related invertebrates.     

Hsps are up-regulated in melanoma tissue and correlate with patient clinical parameters

Heat shock proteins (hsps) have been studied in numerous cancer types, but a clear view of their clinical relevance in melanoma remains elusive. Therefore, the aim of this study was to investigate the expression of hsps in melanoma with respect to patient clinical parameters. Using Western immunoblotting, hsps 90, 70, 60, 40 and 32 were observed to be widely expressed in metastatic melanomas (n = 31), while immunofluorescence demonstrated that in the majority of samples these hsps, apart from hsp32, were increased in expression in melanoma cells compared with surrounding non-melanoma cells in situ (n = 8). Correlating hsp expression with patient clinical parameters indicated that greater hsp90 (P < 0.02) and hsp40 (P < 0.03) expression correlated with advanced stage (stage III Vs stage IV), while in the case of hsp40, this was additionally associated with reduced patient survival (P < 0.05). In contrast, higher hsp32 expression was associated with improved patient survival (P < 0.007). On the other hand, the expression of the other hsps did not correlate with any obtainable patient clinical parameters. This study provides further evidence for the importance of hsps in melanoma and for their use as therapeutic targets and biomarkers, but larger-scale follow-up studies are required to confirm these results.

Electronic supplementary material

The online version of this article (doi:10.1007/s12192-012-0363-1) contains supplementary material, which is available to authorized users.     

Comparative chlorine and temperature tolerance of the oyster crassostrea madrasensis: Implications for cooling system fouling

Crassostrea madrasensis is an important fouling oyster in tropical industrial cooling water systems. C. madrasensis individuals attach to surfaces by cementing one of their two valves to the substratum. Therefore, oyster fouling creates more problems than mussel fouling in the cooling conduits of power stations, because unlike the latter, the shell of the former remains attached to the substratum even after the death of the animal. However, there are no published reports on the tolerance of this species to chlorination and heat treatment. The mortality pattern and physiological behaviour (oxygen consumption and filtration rate) of three size groups (13 mm, 44 mm and 64 mm mean shell length) of C. madrasensis were studied at different residual chlorine concentrations (0.25, 0.5, 0.75, 1, 2, 3 to 5 mg l^{m 1}) and temperatures (30°C to 45°C). The effect of shell size (=age) on C. madrasensis mortality in the presence of chlorine and taking into account temperature was significant, with the largest size group oysters showing highest resistance. At 1 mg l^{m 1} residual chlorine, the 13 mm and 64 mm size group oysters took 504 h (21 d) and 744 h (31 d), respectively to reach 100% mortality. At 39°C, the 13 mm size group oysters took 218 min to reach 100% mortality, whereas the 64 mm size group oysters took 325 min. The oxygen consumption and filtration rate of C. madrasensis showed progressive reduction with increasing residual chlorine concentrations. However, the filtration rate and oxygen consumption responses of C. madrasensis were not significantly different between 30°C (control) and 37.5°C. There was a sharp decrease in the filtration rate and oxygen consumption at 38.5°C. A comparison of the present mortality data with previous reports on other bivalves suggests that the chlorine tolerance of C. madrasensis lies in between that of Perna viridis and Perna perna, while its temperature tolerance is significantly higher than that of the other two bivalve species. However, in power station heat exchangers, where simultaneous chlorine and thermal stresses are existent, C. madrasensis may have an edge over other common foulants, because of its high temperature tolerance.     

Heat shock response in olive (Olea europaea L.) twigs: Identification and analysis of a cDNA coding a class I small heat shock protein

Abstract

The objective of this study was to investigate at the molecular level the heat shock response in olive by identifying sequences coding for heat shock proteins (HSPs). Young twigs of Olea europaea trees (cv Cellina di Nardò) were subjected to different temperature treatments in order to induce the expression of heat shock genes. In order to identify genes induced by heat treatment, we used a PCR-based approach to amplify specific HS cDNAs. Search for low molecular weight HSP sequences was performed in public domain databases in order to design specific primers based upon multisequence alignments. By this approach, we isolated the first full length cDNA encoding a low-molecular weight HSP from O. europaea. This is a class I low molecular weight HSP of 18.3 kDa, which is highly expressed in young twigs subjected to heat stress.     

Carboxyl-terminal Domain of Transient Receptor Potential Vanilloid 1 Contains Distinct Segments Differentially Involved in Capsaicin- and Heat-induced Desensitization

Multiple Ca²⁺-dependent processes are involved in capsaicin-induced desensitization of transient receptor potential vanilloid 1 (TRPV1), but desensitization of TRPV1 by heat occurs even in the absence of extracellular Ca²⁺, although the mechanisms are unknown. In this study, we tested the hypothesis that capsaicin and heat desensitize TRPV1 through distinct mechanisms involving distinct structural segments of TRPV1. In HEK293 cells that heterologously express TRPV1, we found that heat-induced desensitization was not affected by the inclusion of intracellular ATP or alanine mutation of Lys¹⁵⁵, both of which attenuate capsaicin-induced desensitization, suggesting that heat-induced desensitization occurs through mechanisms distinct from capsaicin-induced desensitization. To determine protein domains involved in heat-induced desensitization, we generated chimeric proteins between TRPV1 and TRPV3, a heat-gated channel lacking heat-induced desensitization. We found that TRPV1 with the carboxyl-terminal domain (CTD) of TRPV3 retained heat activation but was impaired in heat-induced desensitization. Further experiments using chimeric or deletion mutants within TRPV1 CTD indicated that the distal half of CTD regulates the activation and desensitization of TRPV1 in modality-specific manners. Within the distal CTD, we identified two segments that distinctly regulated capsaicin- and heat-induced desensitization. The results suggest that the activation and desensitization of TRPV1 by capsaicin and heat can be modulated differentially and disproportionally through different regions of TRPV1 CTD. Identifying the domains involved in thermal regulation of TRPV1 may facilitate the development of novel anti-hyperalgesic approaches aimed at attenuating activation and enhancing desensitization of TRPV1 by thermal stimuli.     

Small area and low connectivity constrain the diversity of plant life strategies in temporary ponds

Aim

(i) To determine whether area and connectivity of temporary ponds can predict plant species diversity, and the diversity and abundance of different plant life histories; (ii) To explore whether pond connectivity with the river prior to river regulation predicts better plant diversity patterns than current pond connectivity, suggestive of possible effects of connectivity loss.

Location

Eastern Carpathian Mountains, Romania, Europe.

Methods

We fitted linear and generalized linear models (LM and GLM) to examine whether pond area and current distance from the Olt River predict plant species richness, Shannon diversity and relative cover of different social behaviour types and overall plant species richness and Shannon diversity. Using historical maps, we measured pond distance from the river ca. 60 years before the Olt River was regulated, and we refitted the LM and GLM models using pond area and past distance from the river as independent variables.

Results

Total plant species richness increased with pond area, and it decreased with the distance from the river, but total plant Shannon diversity index was affected, positively, only by pond area. The strength of responses to pond area and connectivity of species richness, Shannon diversity and relative cover varied across the different social behaviour types. Past and current distances between ponds and riverbeds had similar effects on plant diversity, with some evidence for stronger effect of the present connectivity on specialist species Shannon diversity and a weaker effect on disturbance tolerants, generalists and competitors.

Main Conclusions

Pond area and connectivity with the landscape are important predictors of the diversity of plant life history strategies, and therefore, useful tools in pond conservation. Consistent species richness and Shannon diversity responses of wetland specialists to pond area and connectivity make this life history type well suited for monitoring pond condition.     

Performance limitation and the role of core temperature when wearing light-weight workwear under moderate thermal conditions

The objective of this investigation was to achieve an understanding about the relationship between heat stress and performance limitation when wearing a two-layerfire-resistant light-weight workwear (full-clothed ensemble) compared to an one-layer short sports gear (semi-clothed ensemble) in an exhaustive, stressful situation under moderate thermal condition (25 °C). Ten well trained male subjects performed a strenuous walking protocol with both clothing ensembles until exhaustion occurred in a climatic chamber. Wearing workwear reduced the endurance performance by 10% (p=0.007) and the evaporation by 21% (p=0.003), caused a more pronounced rise in core temperature during submaximal walking (0.7±0.3 vs. 1.2±0.4 °C; p≤0.001) and from start till exhaustion (1.4±0.3 vs. 1.8±0.5 °C; p=0.008), accelerated sweat loss (13±2 vs. 15±3 g min⁻¹; p=0.007), and led to a significant higher heart rate at the end of cool down (103±6 vs. 111±7 bpm; p=0.004). Correlation analysis revealed that core temperature development during submaximal walking and evaporation may play important roles for endurance performance. However, a critical core temperature of 40 °C, which is stated to be a crucial factor for central fatigue and performance limitation, was not reached either with the semi-clothed or the full-clothed ensemble (38.3±0.4 vs. 38.4±0.5 °C). Additionally, perceived exertion did not increase to a higher extent parallel with the rising core temperature with workwear which would substantiate the critical core temperature theory. In conclusion, increased heat stress led to cardiovascular exercise limitation rather than central fatigue.     

Circulating anti-heat-shock-protein antibodies in normal pregnancy and preeclampsia

It has been previously reported that circulating anti-heat-shock-protein (Hsp) antibody levels are elevated in cardiovascular disorders. The aim of the present study was to determine circulating antihuman Hsp60, antimycobacterial Hsp65, and antihuman Hsp70 antibody levels in healthy pregnant women and preeclamptic patients and to investigate their relationship to the clinical characteristics of the study subjects, as well as to the markers of inflammation (C-reactive protein (CRP)), endothelial activation (von Willebrand factor antigen), or endothelial injury (fibronectin), oxidative stress (malondialdehyde) and to serum Hsp70 levels. Ninety-three preeclamptic patients and 127 normotensive healthy pregnant women were involved in this case control study. Serum anti-Hsp60, anti-Hsp65, anti-Hsp70, and Hsp70 levels were measured with enzyme-linked immunosorbent assay (ELISA). Serum CRP levels were determined by an autoanalyzer using the manufacturer’s kit. Plasma von Willebrand factor antigen levels were quantified by ELISA, while plasma fibronectin concentration by nephelometry. Plasma malondialdehyde levels were measured by the thiobarbituric-acid-based colorimetric assay. For statistical analyses, nonparametric methods were applied. Anti-Hsp60, anti-Hsp65, and anti-Hsp70 antibodies were detected in all of our serum samples. There were no significant differences in serum anti-Hsp60, anti-Hsp65, and anti-Hsp70 antibody levels between the control and preeclamptic groups. Serum levels of Hsp70 and CRP, as well as plasma levels of VWF antigen, fibronectin, and malondialdehyde, were significantly higher in preeclamptic patients than in normotensive healthy pregnant women. Serum anti-Hsp60 antibody levels showed significant correlations with serum anti-Hsp65 antibody levels both in the control and the preeclamptic groups (Spearman R = 0.55 and 0.59; p < 0.001, respectively). However, no other relationship was found between clinical features (maternal age, smoking status, parity, body mass index, gestational age at blood draw, systolic and diastolic blood pressure, gestational age at delivery, and fetal birth weight) and measured laboratory parameters of the study subjects and serum anti-Hsp antibody levels in either study group. In conclusion, anti-Hsp60 and anti-Hsp70 antibodies as naturally occurring autoantibodies are present in the peripheral circulation of healthy pregnant women. Nevertheless, humoral immunity against heat shock proteins was not associated with preeclampsia. Further studies are warranted to explore the role of heat shock proteins and immune reactivity to them in the immunobiology of normal pregnancy and preeclampsia.