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BACKGROUND: Heat shock protein 60 (Hsp60) is a chaperonin involved in tumorigenesis, but its participation in tumor development and progression is not well understood and its value as a tumor biomarker has not been fully elucidated. In the current study, the authors presented evidence supporting the theory that Hsp60 has potential as a biomarker as well as a therapeutic target in patients with large bowel cancer. METHODS: The authors studied a population of 97 subjects, including patients and controls. Immunomorphology, Western blot analysis, and quantitative real-time polymerase chain reaction were performed on tissue specimens. Exosomes were isolated from blood and characterized by electron microscopy, biochemical tests, and Western blot analysis. RESULTS: Hsp60 was found to be increased in cancerous tissue, in which it was localized in the tumor cell plasma membrane, and in the interstitium associated with cells of the immune system, in which it was associated with exosomes liberated by tumor cells and, as such, circulated in the blood. An interesting finding was that these parameters returned to normal shortly after tumor removal. CONCLUSIONS: The data from the current study suggested that Hsp60 is a good candidate for theranostics applied to patients with large bowel carcinoma and encourage similar research among patients with other tumors in which Hsp60 has been implicated.

Heat shock proteins (Hsps) assist other proteins in their folding and drive the degradation of defective proteins. During evolution, these proteins have also acquired other roles. Hsp10 is involved in immunomodulation and tumor progression. Hsp90 stabilizes a range of "client" proteins involved in cell signaling. The present study evaluated the expression levels of Hsp10 and Hsp90 in normal mucosa and adenocarcinoma samples of human large bowel.Samples of normal mucosa and adenocarcinoma were collected and Reverse transcriptase-polymerase chain reaction RT-PCR, western blotting (WB) analyses, as well as immunohistochemistry were performed to evaluate the expression levels of Hsp10 and Hsp90.RT-PCR showed a higher gene expression of Hsp10 and Hsp90 in adenocarcinoma samples compared to healthy mucosa. WB results confirmed these findings. Immunohistochemistry revealed higher levels of Hsp10 in adenocarcinoma in both the epithelium and the lamina propria, while Hsp90 expression was higher in the adenocarcinoma samples only in the lamina propria.Hsp10 and Hsp90 may be involved in large bowel carcinogenesis.

Flow boiling heat transfer can provide high heat transfer rates due to latent heat transportation. Its possible use is therefore potentially important to reduce size and weight of cooling systems in space platforms and satellites. A comprehensive knowledge is also important for the safe operation of existing single-phase systems in case of accidental increase of heat generation rate. For space applications and appropriate design of components accommodating flow boiling heat transfer, it is important to understand the influence of microgravity conditions on forced convective boiling heat transfer. The number of existing researches on flow boiling in reduced gravity is very small due to large heat loads required and reduced available room in a 0-g apparatus for experiments, as well as complexity of the experimental facility for microgravity environment. This lecture will summarize the results of the few research carried out on flow boiling heat transfer in microgravity, discussing flow pattern, heat transfer coefficient, critical heat flux, with a particular emphasis to the recent experiments carried out at ENEA. The experiments were carried out at low gravity during the ESA (European Space Agency) parabolic flights campaign of November 2013. The paper will show the analysis of differences between flow patterns and vapour bubble parameters at normal and at zero gravity.

Vasostatin-1 (VS-1), the N-terminal fragment derived from the cleavage of chromogranin A (CgA), has been shown to exert several biological activities on several tissues and organs. Recently, it has been reported that human recombinant VS-1 (STA-CGA(1-78)) may alter myocardial contractility in eel, frog., and rat hearts. In this article we have explored if STA-CGA(1-78) can induce intracellular cascades interacting both with adhesion molecules and/or extracellular matrix (ECM), components, that is, involvement of the heat shock protein 90 (HSP90) and the endothelial NOS (eNOS), known to be implicated in signal transduction mechanisms affecting myocardial contractility. We used 3D cultured adult rat cardiomyocytes cultivated over fibronectin or fibroblasts or embedded in matrigel or collagen type I. Aurion-conjugated VS-1 (AuSTA-CGA(1-78)) has been used to identify possible sites of interaction of thus molecule with the cell membrane. We found that in our 3D culture, cell-ECM interactions played a crucial role in the cellular localization of HSP90 as well as in the expression of eNOS. VS-1 appeared to modulate cell-ECM interactions, thereby remarkably leading to a different cellular localization of HSP90. Moreover., Au-STA-CGA(1-78) was never detected inside the cell nor overlapping the plasma membrane, but nearby the outer side of the cardiomyocyte plasmalemma, at a particular distance, typical of integrins. On the whole, these data suggest that VS-1 does not have a classic receptor on the membrane but that integrins may represent a nonconventional VS-1 receptor modulating eNOS signaling pathway.

Cigarette smoke is a potent stressor for the respiratory system, contributing to pathogenesis, for instance in chronic obstructive pulmonary disease (COPD), but its effects on the expression, function, and cellular localization of mitochondrial chaperonins are still largely unknown. We studied in vivo (airways biopsies) the localization of Hsp10 and Hsp60 in patients (smokers and non-smokers) affected by mild-moderate COPD, and characterized the effects of non-lethal doses of cigarette smoke extract (CSE) on the expression of these molecules in two human cell lines: lung fibroblasts (HFL-1) and bronchial epithelial (16HBE). We applied various in vitro methods: immunohistochemistry (IHC), subcellular fractionation analyses (SFA), Western blotting (WB), immunocytochemistry (ICC), and transmission electron microscopy (TEM) immunogold, and used bioinformatics and databases searches to gather structural in silico data for interpreting and complementing the in vitro results. IHC showed that in smokers and non-smokers COPD patients Hsp10 was localized in both, the cytoplasm and the nucleus of epithelial and lamina propria cells, while Hsp60 was present only in the cytosol. ICC, SFA, and WB on both CSE-exposed cell lines confirmed the presence of nuclear Hsp10, with an increasing trend in parallel to CSE concentration. TEM immunogold further confirmed Hsp10 in the nucleus, in addition to its presence in the cytoplasm and mitochondria, on both cell lines. Bioinformatics and in silico structural analyses indicated that Hsp10 can localize in extramitochondrial sites, such as the nucleus, even if Hsp10 lacks known DNA-binding motifs or nuclear import signals in its primary sequence. Our data suggest a link between exposure to exogenous oxidative stress and cell response, involving Hsp10, which would play roles different from its canonical functions. It is known that Hsp10 can display an array of functions depending on its location: cytoplasm, mitochondria, or extracellular. Here, we show for the first time the presence of Hsp10 in the nucleus of epithelial and stromal human-lung cell lines, paralleling the observations in vivo in COPD patients, and indicating that intranuclear Hsp10 levels are affected by oxidative stress due to an exogenous stressor like cigarette-smoke. The questions now are by what mechanism Hsp10 becomes a resident of the nucleus and what are its functions there.

BACKGROUND: In a previous work we showed for the first time that human tumor cells secrete Hsp60 via exosomes, which are considered immunologically active microvesicles involved in tumor progression. This finding raised questions concerning the route followed by Hsp60 to reach the exosomes, its location in them, and whether Hsp60 can be secreted also via other mechanisms, e.g., by the Golgi. We addressed these issues in the work presented here. PRINCIPAL FINDINGS: We found that Hsp60 localizes in the tumor cell plasma membrane, is associated with lipid rafts, and ends up in the exosomal membrane. We also found evidence that Hsp60 localizes in the Golgi apparatus and its secretion is prevented by an inhibitor of this organelle. CONCLUSIONS/SIGNIFICANCE: We propose a multistage process for the translocation of Hsp60 from the inside to the outside of the cell that includes a combination of protein traffic pathways and, ultimately, presence of the chaperonin in the circulating blood. The new information presented should help in designing future strategies for research and for developing diagnostic-monitoring means useful in clinical oncology.

Molecular chaperones, many of which are heat-shock proteins (HSPs), are an important class of molecules with various functions. Pathological conditions in which chaperones become etiological and/or pathogenic factors are called chaperonopathies, and are classified into by defect, by excess, and by "mistake". In the latter case, the chaperone is structurally and functionally normal but paqrtecipates in pathwais that favor diseases, aòlthough in some cases the chaperone may have post-translational modifications that may lead it to change its location and function and, thus, to become pathogenic. For example, HSP-chaperones are involved in acrcinogenesis in various ways, so that some forms of cancer may be considered "chaperonopathies by mistake". This concept suggests new strategies for anticancer therapy (chaperonotherapy), in which the primary targets or therapeutic agents are choaperones. Chaperonotherapy consists of the utilization of HSP-chaperones for treating chaperonopathies, including cancer. Negative chaperonotherapy is aimed at eliminating or blocking the action of chaperones that favour carcinogenesis or other diseases, whereas positive chaperonotherapy uses chaperones, genes or proteins, to gight against dioseases, such cancer, by stimulating the immune system or the cellular defences against stress.

One of the most important functions of the ear is sound recepction, particularly the detection of amplitude and frequency of the sound waves by Corti's organ. The latter is a sensory structure located in the cochlear duct (middle scale), consisting of hair cells lying above the basilar membrane. The cochlear duct is surrounded by two cavities containing perilymph: the scala vestibularis and the scala tympani. The sound reception mechanism involves several other components of the ear such as malleus, incus and stapes, in the tympanic cavity, and the oval and round windows. The movement of the stapes on the oval window, that is adjacent to the scala vestibularis, generates pressure waves in the perilymph along the vestibular canal. The round window, that separates the scala tympani from the tympanic cavity, moves to compensate for oval window movements. The malfunction of the inner ear, due to specific diseases, could be corrected by use of the drugs such as gentamicin that reaches the cochlea through the round window. This brief paper reviews the main anatomical knowledge on the inner ear, with particular attention to the structures of interest for otolaryngologists treating ear disorders by inoculation of drugs into the tympanic cavity.

Cigarette smoke is a potent stressor for the respiratory system, contributing to pathogenesis, for instance in chronic obstructive pulmonary disease (COPD), but its effects on the expression, function, and cellular localization of mitochondrial chaperonins are still largely unknown. We studied in vivo (airways biopsies) the localization of Hsp10 and Hsp60 in patients (smokers and non-smokers) affected by mild-moderate COPD, and characterized the effects of non-lethal doses of cigarette smoke extract (CSE) on the expression of these molecules in two human cell lines: lung fibroblasts (HFL-1) and bronchial epithelial (16HBE). We applied various in vitro methods: immunohistochemistry (IHC), subcellular fractionation analyses (SFA), Western blotting (WB), immunocytochemistry (ICC), and transmission electron microscopy (TEM) immunogold, and used bioinformatics and databases searches to gather structural in silico data for interpreting and complementing the in vitro results. IHC showed that in smokers and non-smokers COPD patients Hsp10 was localized in both, the cytoplasm and the nucleus of epithelial and lamina propria cells, while Hsp60 was present only in the cytosol. ICC, SFA, and WB on both CSE-exposed cell lines confirmed the presence of nuclear Hsp10, with an increasing trend in parallel to CSE concentration. TEM immunogold further confirmed Hsp10 in the nucleus, in addition to its presence in the cytoplasm and mitochondria, on both cell lines. Bioinformatics and in silico structural analyses indicated that Hsp10 can localize in extramitochondrial sites, such as the nucleus, even if Hsp10 lacks known DNA-binding motifs or nuclear import signals in its primary sequence. Our data suggest a link between exposure to exogenous oxidative stress and cell response, involving Hsp10, which would play roles different from its canonical functions. It is known that Hsp10 can display an array of functions depending on its location: cytoplasm, mitochondria, or extracellular. Here, we show for the first time the presence of Hsp10 in the nucleus of epithelial and stromal human-lung cell lines, paralleling the observations in vivo in COPD patients, and indicating that intranuclear Hsp10 levels are affected by oxidative stress due to an exogenous stressor like cigarette-smoke. The questions now are by what mechanism Hsp10 becomes a resident of the nucleus and what are its functions there., Italian Journal of Anatomy and Embryology, Vol 118, No 2 (Supplement) 2013

It is currently accepted that the human Hsp60 resides and works not only in the mitochondria, the canonical residence, but also outside it. It is also known that Hsp60 although coded by a nuclear gene is synthesized in the cytosol and includes an N-terminal mitochondrial import signal (MIS), which directs the polypeptide toward the inside of the organelle where the MIS is removed. Therefore, there are at least two functional types of Hsp60, with and without MIS, the former in the cytosol the latter inside the mitochondria. A key question is: how do these two forms of Hsp60 differ beyond the fact that while one has MIS the other lacks it? How presence or absence of MIS affects the ability of Hsp60 to form oligomers, which are considered important for chaperoning peptides and assist them to reach a native state? We report here our initial observations on this issue. Typically, in the mitochondria, Hsp60 forms ring-shaped heptamers, two of which associate to build a barrel-shaped tetradecamer, the functional chaperoning complex. It is not known if the cytosolic Hsp60 with its MIS, also forms hepta- and tetradecamers. A clarification of this issue will most likely shed light on the physiological functions of extramitochondrial Hsp60, and also on its pathogenic role in Hsp60 chaperonopathies. Consequently, we compared recombinant Hsp60 bearing the MIS with the prokaryotic ortholog GroEL, which under normal conditions forms functional double-ring tetradecamers. Characteristic hydrodynamic sizes of the oligomeric complex for both systems were investigated by small angle X-ray (SAXS) and static and dynamic light scattering (SLS and DLS) in solution under similar physicochemical conditions. High Performance Liquid Chromatography (HPLC) and blue native polyacrylamide-gel electrophoresis were used to further clarify the equilibrium between the different oligomeric species of the two proteins over a wide range of concentrations. Hsp60 with MIS formed hepta- and tetradecamers similarly to GroEL. Oligomerization was dependent on concentration for GroEL and Hsp60, but for the latter, formation of larger oligomers, e.g., tetradecamers, required higher concentrations than the former., Italian Journal of Anatomy and Embryology, Vol 118, No 2 (Supplement) 2013

In this paper will be described an experimental campaign aimed to determine the experimental values of the Nusselt number for rough microtubes having a relative roughness up to 4% in laminar and transitional regime. Three microtubes having an inner diameter equal to 440, 280 and 146 m uniformly heated by Joule effect with a constant power DC supply have been tested. The experimental results have evidenced that in laminar regime the average Nusselt number approaches the fully developed constant for uniformly heated tubes when the Reynolds number decreases since the axial conduction along the fluid and the conjugate heat transfer to the walls can be neglected at low Reynolds numbers for the operative conditions adopted during the tests. For higher Reynolds numbers the region of thermal development increases its importance, augmenting the average convective heat transfer coefficient. It has been demonstrated that large values of the microtube roughness influence more strongly the friction factor than the Nusselt number. The experimental results obtained in this work demonstrate that in microtubes having a relative roughness less than 5% the effect of the roughness on the Nusselt number can be considered negligible.