kD Heat Shock Protein Research Articles

Structural and functional abnormalities of hepatic tissues in male Wistar rats fed hyperwhey and super amino anabolic protein

ObjectiveAthletes and bodybuilders consume high-protein supplements to obtain energy and enhance the development and strength of their muscles. Over time, different investigations have revealed dysfunctions of their body organs. There are contradictions among scientists concerning the benefits and the alarm of developing body dysfunction. The aim of this study was to illustrate the effects on consumption of two anabolic protein supplements on body weight and structure and function of hepatocytes in male albino Wistar rats. MethodsWe assigned male Wistar albino rats into three groups (n = 10 each): control, hyperwhey protein (Nutrabolics, Richmond, Canada) (2.5 g/kg body weight), and super amino 2500 (SA) (APN, Ft. Launderale, FL, USA) (2.5 g/kg body weight). The applied dose was orally administered daily in tap water for 14 wk. Body weight was regularly measured. At 14 wk, animals were sacrificed and dissected. Blood was collected from a puncture of the heart and the liver was removed and weighed. Biochemical analysis of liver function tests, lipidogram, hematology, histopathology, transmission electron microscopy, immunohistochemistry of proliferating cell nuclear antigen, B-cell lymphoma 2 and 70 kd heat shock proteins, and flow-cytometry of hepatocyte cell cycle were performed. ResultsHyperwhey- and SA-supplemented rats had lower body weight gain compared with the control group and developed hepatic dysfunction manifested by apparent congestion of blood vessel, increased apoptosis, and breakdown of hepatocytes. The SA group had thickening of the liver capsule and more drastic damage of hepatocytes. The level of transaminases was markedly increased. Insulin level was also markedly decreased in parallel with increase cholesterol, low-density lipoprotein, and triacylglycerols. ConclusionHyperwhey and SA protein formula administration dramatically altered the liver function and increased hepatic damage similar to the development of suspected diabetes.

Somatostatin Reduces the Acute Lung Injury of Mice via Increasing the Affinity of Glucocorticoid Receptor

Background/Aims: Although it has been reported that somatostatin (SOM) upregulated the level of 90-kD heat shock protein (Hsp90), which participates in the inflammatory regulation by its client proteins, such as glucocorticoid receptor (GR), it remains unclear if it has a protective role against acute lung injury (ALI). Methods: ALI model was established by the injection of oleic acid (OA) into the tail vein of mice. Lung injury was assessed by histological analysis, lung water content and arterial blood gases. The levels of Hsp90 and GR, the binding capacity and the affinity of GR were examined. Results: It was showed that pretreatment with SOM significantly increased Hsp90 levels and alleviated lung injuries in OA-injected mice. Furthermore, SOM increased the GR expression and improved the affinity of the GR in animals with lung injury. However, little alteration was found in the maximum binding capacity of the GR in mice with or without SOM. Conclusion: The data indicate SOM exerts a protective effect by increasing Hsp90 abundant and further enhancing the affinity of the GR. The beneficial effects of SOM treatment provide a new strategy for modulation of GR efficiency and alleviation of acute lung injury.

Overexpression of MuHSP70 gene from Macrotyloma uniflorum confers multiple abiotic stress tolerance in transgenic Arabidopsis thaliana.

A 70-KD heat shock protein (HSP70) is one of the most conserved chaperones. It is involved in de novo protein folding and prevents the aggregation of unfolded proteins under lethal environmental factors. The purpose of this study is to characterise a MuHSP70 from horsegram (Macrotyloma uniflorum) and elucidating its role in stress tolerance of plants. A MuHSP70 was cloned and characterised from a natural drought stress tolerant HPK4 variety of horsegram (M. uniflorum). For functional characterization, MuHSP70 was overexpressed in transgenic Arabidopsis. Overexpression of MuHSP70 was found to provide tolerance to the transgenic Arabidopsis against various stresses such as heat, cold, drought, salinity and oxidative stress. MuHSP70 transgenics were observed to maintain the shoot biomass, root length, relative water content, and chlorophyll content during exposure to multi-stresses relative to non-transgenic control. Transgenic lines have further shown the reduced levels of MDA, H2O2, and proteolytic activity. Together, these findings suggest that overexpression of MuHSP70 plays an important role in improving abiotic stress tolerance and could be a crucial candidate gene for exploration in crop improvement program.

Attenuation of experimental asthma by mycobacterial protein combined with CpG requires a TLR9-dependent IFN-γ-CCR2 signalling circuit.

Allergic asthma is a chronic pulmonary disease characterized by a Th2 inflammatory response. The modulation of a Th2 immune response based on immune deviation to a Th1 pattern or induction and migration of regulatory T cells to the lungs constitutes one of the major therapeutic approaches that is being investigated for the treatment of allergic asthma. The potentials of Mycobacterium leprae 65-kD heat-shock protein or Toll-like receptor 9 ligand (CpG oligodeoxynucleotides) as immune modulators for the treatment of airway allergic disease have been studied individually. Mycobacterial protein combined with CpG was used as immunotherapy for airway allergy. Using an ovalbumin-induced asthma model, mice were sensitized and challenged, and then treated with mycobacterial heat-shock protein (Hsp65) combined with CpG. The treatment of mice with established allergy led to the attenuation of eosinophilia, Th2 cytokines and airway hyperresponsiveness. Hsp65 plus CpG treatment also induced an increase in OVA-specific IFN-γ levels and in the frequency of lung inflammatory monocytes. Moreover, we show that the reduction of eosinophilia and the recruitment of inflammatory monocytes to the lungs required early triggering of TLR9, IFN-γ and CCR2 by immunotherapy components. In addition to immune deviation to a Th1 response in the modulation of Th2 allergic inflammation, our findings also attribute an important role to the innate response mediated by TLR9, associated with the recruitment of CCR2-dependent monocytes. Our findings show that the Hsp65/CpG treatment is a promising strategy for consideration in translational studies.

Naturally Occurring Single Nucleotide Mutations Alter the Function of a Highly Conserved Hsp70 Gene in Humans

Molecular chaperones and in particular the 70‐kD heat shock proteins (Hsp70s) are key regulators of the cellular stress response and alterations in their function have direct physiological consequences. To determine how natural mutations alter the function of Hsp70s we used single nucleotide polymorphisms (SNPs) found on HSPA1A the major stress inducible Hsp70 gene in humans. Specifically, the wild‐type (WT) HSPA1A sequence was subcloned into both bacterial and mammalian expression vectors and the mutated gene variants were generated using site directed mutagenesis. Recombinant proteins corresponding to the WT and mutated variants were then generated and tested for their ability to hydrolyze ATP. These experiments revealed that these mutations significantly change the rate by which the protein hydrolyzes ATP. The mutant proteins' ability to properly localize intracellularly within mammalian cells was also tested using immunofluorescence and confocal microscopy. These experiments demonstrated that the mutants and the WT protein had similar subcellular localization. However, live‐dead and cell proliferation assays revealed that some of these mutations significantly change cellular survival after heat stress. Given that these natural variants are either population‐specific or clinical we believe that the observed functional differences alter the ability of cells and the individuals carrying them to cope with stress or disease.

Reproduction reduces HSP70 expression capacity in Argopecten purpuratus scallops subject to hypoxia and heat stress

In scallops, gonad production is highly demanding energetically, and reproduction usually occurs during springsummer, a period of strong environmental changes. The synthesis of heat-shock proteins (HSPs) is a major mechanism of stress tolerance in animals, including scal- lops, and HSP expression contributes considerably to cellular energy demand. Therefore, repro- ductive investment may limit the availability of energy (in terms of ATP) for the expression of HSP in organisms exposed to environmental stress. We evaluated the stress response capacity of adult Argopecten purpuratus scallops to high temperature and hypoxia. Stress response capacity was assessed through gene expression (for temperature stress) and protein induction of 70 kD HSP at 3 reproductive stages: immature, mature and spawned. We also evaluated the effect of reproduc- tive status on the cellular ATP provisioning capacity through citrate synthase activity. Immature scallops exposed to thermal stress showed 1.3- and 1.5-fold increases in hsp70 mRNA and HSP70 protein levels, respectively, and those exposed to hypoxia doubled their level of HSP70 compared to non-stressed immature scallops. However, following gonad maturation and spawning, hsp70 mRNA increased by only 0.49- and 0.65-fold, respectively, after thermal stress and HSP70 protein levels of scallops exposed to thermal and hypoxia stressors did not differ from those of non- stressed animals. In parallel, citrate synthase showed its highest level in immature scallops, declined with gonad maturation, and was lowest in spawned scallops. These results suggest that reproductive investment reduces the stress response capacity of A. purpuratus and that mature and spawned scallops could be more vulnerable to environmental stressors than immature individuals.

Prediction of allosteric sites on protein surfaces with an elastic-network-model-based thermodynamic method.

Allostery is a rapid and efficient way in many biological processes to regulate protein functions, where binding of an effector at the allosteric site alters the activity and function at a distant active site. Allosteric regulation of protein biological functions provides a promising strategy for novel drug design. However, how to effectively identify the allosteric sites remains one of the major challenges for allosteric drug design. In the present work, a thermodynamic method based on the elastic network model was proposed to predict the allosteric sites on the protein surface. In our method, the thermodynamic coupling between the allosteric and active sites was considered, and then the allosteric sites were identified as those where the binding of an effector molecule induces a large change in the binding free energy of the protein with its ligand. Using the proposed method, two proteins, i.e., the 70 kD heat shock protein (Hsp70) and GluA2 alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor, were studied and the allosteric sites on the protein surface were successfully identified. The predicted results are consistent with the available experimental data, which indicates that our method is a simple yet effective approach for the identification of allosteric sites on proteins.

Inhibition of adjuvant-induced arthritis by nasal administration of novel synthetic peptides from heat shock protein 65.

BackgroundRheumatoid arthritis (RA) is a chronic systemic inflammatory disease mediated by T cells. The aim of the present study was to investigate the therapeutic efficacy of synthetic peptides (HP-R1, HP-R2 and HP-R3), derived from the sequence of 65-kD mycobacterial heat shock protein (HSP), in the treatment of RA using adjuvant-induced arthritis (AA) animal model.MethodsAA was induced by a single intradermal injection Freund’s complete adjuvant in male Lewis rats. At the first clinical sign of disease, rats were administered nasally by micropipette of peptides or phosphate buffer saline (PBS). Disease progression was monitored by measurement of body weight, arthritis score and paw swelling. The changes of histopathology were assessed by hematoxylin eosin staining. The serum levels of tumor necrosis factor (TNF) - alpha and interleukin (IL)-4 were measured by enzyme-linked immunosorbent assay (ELISA).ResultsThe peptides efficiently inhibited the footpad swelling and arthritic symptoms in AA rats. The synthetic peptides displayed significantly less inflammatory cellular infiltration and synovium hyperplasia than model controls. This effect was associated with a suppression of pro-inflammatory cytokine TNF-alpha production and an increase of anti-inflammatory cytokine IL-4 production after peptides treatment.ConclusionsThese results suggest that the synthetic peptides derived from HSP65 induce highly effective protection against AA, which is mediated in part by down-regulation of inflammatory cytokines, and support the view that the synthetic peptides is a potential therapy for RA that may help to diminish both joint inflammation and destruction.

SAT0274 IL-9-Producing TH 9 Cells May Participate in Pathogenesis of Takayasu's Arteritis

BackgroundTakayasu's arteritis (TA) is a granulomatous large vessel vasculitis predominantly affecting the aorta and its major division branches. Over the past decades, cell-mediated autoimmunity has been implicated in the pathogenesis...

Male immunity to the chlamydial 60 kDa heat shock protein (HSP 60) - associated with semen quality?

The role of Chlamydia trachomatis for male infertility is a matter of constant debate. It is assumed that in its persistent form this pathogen may produce high levels of 60 kD heat shock protein (Chlam HSP60). Cross-reactivity between epitopes of the bacterial and human HSPs, involved in many steps of the reproductive process, might induce an autoimmune response with potential impairment of semen quality and sperm fertilising capacity. This prospective study included asymptomatic males of a total of 128 unselected subfertile couples (median duration of infertility 3 years) to determine the clinical relevance of male immunity to Chlam HSP60 during infertility investigation. After medical history and clinical examination of both partners, serum antibodies (Ab) to Chlam HSP60 were determined. Same day semen quality evaluation included microscopical standard sperm analysis, determination of seminal white blood cells (WBC) and of antisperm Ab (ASA) of the Ig G- and Ig-A class (mixed antiglobulin reaction, MAR), microbial screening and examination of sperm functional capacity. Sperm/mucus interaction was tested in vitro and in vivo. Simultaneously, patients' female partners were tested for Chlam HSP60 Ab and results were compared with a standard serology evaluation for antichlamydial IgG Ab. The presence of ChlamHSP60 Ab (positive in 24% of males) was not significantly associated with semen quality, seminal WBC and antisperm AB of the IgG- or Ig A-class, the outcome of the microbial screening nor with sperm functional capacity and results of sperm/mucus interaction testing in vitro and in vivo. Chlam HSP60 Ab were significantly more frequent in female partners of Chlam HSP60 Ab-positive men, and results correlated with the outcome of standard chlamydial serology evaluation. In conclusion, when serum Chlam HSP60 Ab are used as marker, male immunity to the chlamydial 60 kD heat shock protein is not associated with semen quality, sperm functional capacity and other clinically relevant parameters of male fertility.

Allosteric Opening of the Polypeptide-Binding Site When an Hsp70 Binds ATP

The 70 kD heat shock proteins (Hsp70s) are ubiquitous and highly conserved molecular chaperones essential for cellular protein folding and proteostasis. Each Hsp70 has two functional domains: a nucleotide-binding domain (NBD) that binds and hydrolyzes ATP, and a substrate-binding domain (SBD) that binds extended polypeptides. NBD and SBD interact little when in ADP; however, ATP binding allosterically couples the polypeptide- and ATP-binding sites. ATP binding promotes polypeptide release; polypeptide rebinding stimulates ATP hydrolysis. This allosteric coupling was poorly understood. To explore the molecular mechanism of this essential ATP-induced allosteric coupling, we solved a crystal structure of an intact Hsp70 from E. coli in complex with ATP at 1.96 A resolution. NBD-ATP adopts a unique conformation, forming extensive interfaces with a radically changed SBD that has its α-helical lid displaced and the polypeptide-binding pocket of its β-subdomain flipped open. Our biochemical analysis inspired by this structure provides a long-sought mechanistic explanation of how ATP binding allosterically opens the polypeptide-binding site.

Innate and Adaptive Responses to Heat Shock Proteins in Behcet’s Disease

Behcet's disease (BD) is a systemic, chronic inflammatory disorder with both innate and adaptive immune responses. Heat shock proteins (HSP) are highly conserved molecules in different species with scavenger activity and involved in correct folding of newly synthesized proteins. T and B cell responses against HSPs are observed in BD patients in both αβ and γδ T-cell populations. 60-kD HSP (HSP60) is also shown to be recognized by pattern recognition receptors such as toll-like receptors (TLR) and is suggested to be an endogenous “danger” signal to the immune system with rapid inflammatory cytokine releases and enhancement of adaptive Th1-type responses. Elucidating the exact role of HSPs in BD pathogenesis might pave the way to less toxic therapeutic approaches to BD, such as antibacterial therapies and immunomodulation.

Molecular characteristics and expression analysis of ScHsc70 cDNA in agamaki clam (Sinonovacula constricta)

Heat shock proteins consist of several families of highly conserved proteins that play an essential role in a number of cellular processes.Among the 70 kD family of heat shock proteins,heat shock cognate protein 70 kD(Hsc70) and inducible heat shock protein 70 kD(Hsp70) have been extensively studied in vertebrates and invertebrates.Several cDNAs encoding HSP70 have been described in molluscs,including the oyster(Crassostrea gigas),bay scallop(Argopecten irradians),Zhikong scallop(Chlamys farreri),mussel(Mytilus galloprovincialis),and Asiatic hard clam(Meretrix meretrix).Recent studies examining HSP70 expression in different species of mollusc have recognized the physiological and ecological importance of heat-shock gene expression in response to changing environments.We isolated an EST sequence with high homology with heat shock protein 70 gene from the cDNA library of Sinonovacula constricta.Then,the complete express sequence of this gene was obtained using PCR and 5′RACE.The cDNA of this gene was 2 335 bp,and consisted of a 76 bp 5′ untranslated region(UTR),a 1 950 bp open reading frame(ORF) and a 309 bp 3′ UTR.The translated protein consisted of 649 amino acids(70.89 kD)and its calculated isoelectric point was 5.28.Sequence analysis of the protein revealed that this gene contained three signature sequences of the heat shock protein 70 family(HSP70 family),two glycosylation sites and one ATP-GTP binding site.Four terapeptides of GGXP and a cytoplasm characteristic motif of EEVD were detected in the carboxyl terminal region of the deduced amino acid sequence.This HSP70 is a member of the HSC70(constitutive genes) subfamily in the HSP70 family,and is designated as ScHsc70.Phylogenetic analysis suggested that the protein was most similar to those of S.constricta,Laternula elliptica,and M.meretrix.Quantitative reverse transcriptase(qRT-PCR) analyses revealed that ScHsc70 mRNA was expressed constitutively in all the tissues tested.Expression was lowest in the mantle and highest in the water pipe and liver.Following challenge with Vibrio parahaemolyticus and V.anguillarum,ScHsc70 mRNA expression first increased in the liver of S.constricta then decreased.Expression peaked after 20 and 30 h post-challenge with V.parahaemolyticus and V.anguillarum,respectively.There was a significant increase in ScHsc70 mRNA in the liver of clams challenged with V.parahaemolyticus compared to the control group at 4,12,and 30 h post-challenge.Similarly,there was a significant increase in ScHsc70 mRNA levels in the liver of clams challenged with V.anguillarum 4,8,12,20,30,and 48 h post-challenge.Our results suggest that ScHsc70 may play a role in the immune response of S.constricta to bacterial challenge.

Immunohistochemical investigation of the coma blister and its pathogenesis

The erythematous patches and vesicles that are observed in coma patients, usually from an overdose of medication, are known as coma blisters. However, it is unknown whether the degenerated sweat gland is a necrosis or apoptosis. We immunohistochemically examined such skin lesions to investigate the characteristics and pathogenesis of the coma blister. Skin lesions were obtained from a forensic autopsy case, a woman in her thirties, of caffeine intoxication. Those lesions were observed in the left femoral, the lower left thigh, and the right knee. Histologically, the skin lesions showed that the keratinocytes had necrosed and the epidermis was thin in some areas. Eccrine sweat gland degeneration was observed. Obvious inflammatory cell infiltrations were not detected. Immunohistochemically, we stained each skin lesion against CD3, CD8, CD45RO, cytokeratin, 70 kD heat shock protein, ubiquitin, 150 kD oxygen regulated protein, and caspase-cleaved keratin 18 neo-epitope M30. They were also stained with an in situ apoptosis detection kit. Degenerated sweat glands featured CD45RO and M30 immunoreactivity. Immunohistochemical staining for CD45RO, CK-L, and M30 might be useful to observe sweat gland degeneration in the coma blister. Therefore, the apoptosis might be related to coma blisters and sweat gland degenerations.

Daily and seasonal changes in heat exposure and the Hsp70 level of individuals from a field population of Xeropicta derbentina (Krynicki 1836) (Pulmonata, Hygromiidae) in Southern France.

The Mediterranean land snail Xeropicta derbentina forms huge populations in Southern France. In order to characterize heat exposure and the induction of the 70-kD heat shock protein (Hsp70) response system during the life cycle of this snail, a selected population from the Vaucluse area, Provence, was investigated encompassing the issues of morphological life cycle parameters (shell size and colouration), the daily courses of heat exposure at different heights above the ground, of shell temperature, and that of the individual Hsp70 levels. The study covered all four seasons of the year 2011. Snails were found to be annual, reaching their final size in August. The shell colouration pattern showed high variation in juveniles (spring) with a strong tendency towards becoming uniformly white at old age in autumn. In all seasons, ambient air temperature decreased with increasing distance from the ground surface during daytime while remaining constantly low in the night. Overall, the Hsp70 level of individuals followed the ambient temperature during diurnal and seasonal variations. Correlation analysis revealed a positive association of individual shell temperature and Hsp70 level for the most part of the life cycle of the snails until late summer, whereas a negative correlation was found for aged animals indicating senescence effects on the capacity of the stress response system.

Proteomic identification of proteins relevant to ovarian development in the red swamp crayfish Procambarus clarkii

In this study, the two-dimensional gel electrophoresis patterns of total proteins from previtellogenic (stage III) and vitellogenic (stage IV) ovaries of Procambarus clarkii were compared for the first time. A total of 52 significantly differentially expressed protein spots (up/down-regulated≥2 fold) were excised for further analysis using matrix-assisted laser desorption/ionization-tandem time of flight mass spectrometry. There were 29 spots successfully identified, including 22 unique proteins. Compared with stage III ovaries, 13 proteins in the stage IV ovaries were down-regulated (e.g., tropomysin, 70kD heat shock protein, protein disulfide-isomerase A6, glutathione S transferase D1, arginine kinase, and Na+/K+-ATPase) and 9 were up-regulated (e.g., peritrophin, cyclin B, elongation factor 2, transketolase, checkpoint kinase 1, and I-connectin). The results indicated the potential of cellular proteomic studies for the identification of proteins functionally involved in the ovarian development of P. clarkii.

Identification of HnRNP-A2/B1 as a Target Antigen of Anti-Endothelial Cell IgA Antibody in Behçet's Disease

Behçet's disease (BD) is a chronic, multisystemic vasculitis that theoretically affects all sizes and types of blood vessels. Although pathogenesis remains enigmatic, endothelial cells are believed to be the primary target in this disease. We detected the target protein using western blotting and immunoprecipitation and determined the amino-acid sequence of the peptide by liquid chromatography-matrix assisted laser desorption/ionization-tandem time-of-flight analysis (LC-MALDI-TOF/TOF). Serum reactivity against the recombinant target protein was analyzed by immunoblotting. Serum reactivity against streptococcal 65-kD heat shock protein (hsp-65) and the recombinant target protein was investigated by ELISA. The 36-40-kD protein band that was obtained from immunoprecipitation, which was analyzed by LC-MALDI-TOF/TOF, exhibited the amino-acid sequences of heterogeneous nuclear ribonucleoproteins A2/B1 (hnRNP-A2/B1). Reactivity of serum IgA against human recombinant hnRNP-A2/B1 was detected in 25 of 30 BD patients (83.3%), 4 of 30 systemic lupus erythematosus patients (13.3%), 8 of 30 rheumatoid arthritis patients (26.7%), 9 of 30 Takayasu's arteritis patients (30%), 6 of 30 healthy controls (20%), and none of 30 IgA nephropathy patients. Optical densities obtained from ELISAs against the recombinant human hnRNP-A2/B1 were correlated with those against the recombinant streptococcal hsp-65.JID JOURNAL CLUB ARTICLE: For questions, answers, and open discussion about this article, please go to http://www.nature.com/jid/journalclub.

Comparison of epitope specificity of anti-heat shock protein 60/65 IgG type antibodies in the sera of healthy subjects, patients with coronary heart disease and inflammatory bowel disease

Previously, we reported on the presence of antibodies to linear epitopes of human and mycobacterial 60 kD heat shock proteins (HSP) in the sera of healthy blood donors. Since many recent findings indicate that the levels of these antibodies may be altered in coronary heart disease (CHD) and also inflammatory bowel diseases (IBD), it seemed worthwhile to compare the epitope specificity of the anti-HSP60 and anti-HSP65 antibodies in the sera of patients with these diseases to those in healthy subjects. The multipin enzyme-linked immunosorbent assay method was applied with a large overlapping set of synthetic 10-mer peptides covering selected regions of human HSP60 and Mycobacterium bovis HSP65. Sera of 12 healthy persons (HP), 14 CHD, and 14 IBD patients with the same concentration of total anti-HSP60 and HSP65 IgG antibodies were tested. We have identified CHD-specific epitopes in the equatorial domain of the HSP60 protein but in neither region of the HSP65 molecule, indicating that the formation of anti-HSP60 antibodies is not or only partially due to the cross-reaction between human HSP60 and bacterial HSP65. IBD-specific epitopes were found in many regions of the HSP60 and in even more regions of the HSP65 molecule including an IBD-specific T cell epitope in region X as well. These findings indicate that the epitope specificity of the anti-human and anti-mycobacterial HSP60 antibodies associated with various diseases is different.

Heat Shock Protein 47 (HSP47) Antisense Oligonucleotides Reduce Cardiac Remodeling and Improve Cardiac Function in a Rat Model of Myocardial Infarction

Cardiac remodeling after acute myocardial infarction is regulated by components of the extracellular matrix. The 47 kD heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone that plays a major role during procollagen processing and/or secretion. The purpose of the study was to determine whether HSP47 inhibition can mitigate ligated left anterior descending (LAD) coronary artery-induced myocardial infarction in rats. Rats were randomly divided into four experimental groups and subjected to the following treatments: 1) intravenous (IV) administration of saline; 2) ligation of the LAD coronary artery; 3) ligation of the LAD coronary artery + IV administration of HSP47 antisense oligonucleotides; or 4) IV administration of HSP47 antisense oligonucleotides. We investigated cardiac histopathology, performed immunoblot and immunohistochemical analyses, and examined cardiac function. Rats with ligated LAD coronary artery experienced upregulation of HSP47 expression, remodeling of the left ventricle, and cardiac dysfunction. In contrast, rats with ligated LAD coronary artery treated with HSP47 antisense oligonucleotides had significantly reduced HSP47 expression, cardiac remodeling, and improved cardiac function. Intravenous (IV) administration of HSP47 antisense oligonucleotides alone had no effect on cardiac morphology. The data strongly support the idea that changes in the extracellular matrix and its components are important determinants of cardiac remodeling after myocardial infarction.

Behçet's Disease (Adamantiades-Behçet's Disease)

Adamantiades-Behçet's disease (ABD) is characterized by starting with oral aphthous ulceration and developing of the systemic involvements. The pathogenesis of ABD is closely correlated with the genetic factors and the triggering factors which acquire delayed-type hypersensitivity reaction against oral streptococci mediated by IL-12 cytokine family. HLA-B51 is associated in more than 60% of the patients and its restricted CD8+ T cell response is clearly correlated with the target tissues. Bes-1 gene encoded partial S. sanguinis genome which is highly homologous with retinal protein, and 65 kD heat shock protein (Hsp-65) released from streptococci is playing an important role with human Hsp-60 in the pathogenesis of ABD. Although Hsp-65/60 has homologies with the respective T cell epitope, it stimulates peripheral blood mononuclear cells (PBMCs) from ABD patients. On the other hand, some peptides of Hsp-65 were found to reduce IL-8 and IL-12 production from PBMCs of ABD patients in active stage.