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Einstein (Säo Paulo) ; 20: eRB6181, 2022. tab
Article in English | LILACS | ID: biblio-1364795


ABSTRACT Ischemia-reperfusion injury is a pathophysiological event occuring after abdominal organ transplantation, and has a significant influence on prognosis and survival of the graft. It is involved in delaying the primary function or non-functioning of the graft. The objective of this study was to provide information on heat shock protein mechanisms in ischemia-reperfusion injuries in abdominal organ transplantations, and to indicate the possible factors involved that may influence the graft outcome. Several classes of heat shock proteins are part of the ischemia and reperfusion process, both as inflammatory agonists and in protecting the process. Studies involving heat shock proteins enhance knowledge on ischemia-reperfusion injury mitigation processes and the mechanisms involved in the survival of abdominal grafts, and open space to support therapeutic future clinical studies, minimizing ischemia and reperfusion injuries in abdominal organ transplantations. Expression of heat shock proteins is associated with inflammatory manifestations and ischemia-reperfusion injuries in abdominal organ transplantations and may influence graft outcomes.

Reperfusion Injury , Organ Transplantation , Heat-Shock Proteins/metabolism , Ischemia
Int. j. morphol ; 38(5): 1455-1462, oct. 2020. tab, graf
Article in English | LILACS | ID: biblio-1134462


SUMMARY: This study aimed to investigate the changes in testis tissue of thioacetamide-induced rats and the effect of melatonin on these changes. Thirty-five male Wistar Albino rats were divided into five groups. Group I; Control (n=7), Group II; Melatonin (Mel) (10 mg/kg) a single dose (i.p)(n=7), Group III; Thioacetamide (TAA) (300 mg/kg) (i.p) 2 times with 24 hour intervals (n=7), Group IV; TAA (300 mg/kg) was administered at 24-hour intervals, afterwards of 10 mg/kg single dose of Mel (n=7), Group V; Mel was administered 10 mg/kg a single dose 24 hours before the administration of TAA (n=7). Testis was evaluated histologically, immunohistochemically (Heat Shock Proteins (HSP) 70 and 90), blood serum testosterone, total antioxidant status(TAS) and total oxidant status(TOS) in tissue. The tissue sections of Group III decreased seminiferous tubule diameters, and germinal epithelium spills were observed. HSP70 and HSP90 expressions were increased. There wasn't a statistically significant change in testosterone levels among the groups. While TAS levels decreased in Group III compared to control, TOS levels didn't change. HSP70 and HSP90 decreased in groups with Mel-treated. Mel was found to have both protective and therapeutic effects. According to our results, the therapeutic effect of Mel in thioacetamide-induced acute testicular injury is greater than its protective effect.

RESUMEN: Este estudio tuvo como objetivo investigar los cambios en el tejido testicular de ratas inducidas por tioacetamida y el efecto de la melatonina en estos cambios. Treinta y cinco ratas macho Wistar Albino se dividieron en cinco grupos. Grupo I; Control (n = 7), Grupo II; Melatonina (Mel) (10 mg / kg) una dosis única (i.p) (n = 7), Grupo III; Tioacetamida (TAA) (300 mg / kg) (i.p) 2 veces con intervalos de 24 horas (n = 7), Grupo IV; TAA (300 mg / kg) se administró a intervalos de 24 horas, luego de una dosis única de 10 mg / kg de Mel (n = 7), Grupo V; Mel recibió 10 mg / kg de una dosis única 24 horas antes de la administración de TAA (n = 7). Los testículos se evaluaron histológicamente, inmunohistoquímicamente (proteínas de choque térmico (PCT) 70 y 90), testosterona en suero sanguíneo, estado antioxidante total (EAT) y estado oxidante total (EOT) en el tejido. En secciones de tejido del Grupo III se observó disminución de los diámetros de los túbulos seminíferos y derrames en el epitelio germinal. Se aumentaron las expresiones HSP70 y HSP90. No hubo un cambio estadísticamente significativo en los niveles de testosterona entre los grupos. Mientras que los niveles de EAT disminuyeron en el Grupo III en comparación con el control, los niveles de EOT no cambiaron. HSP70 y HSP90 disminuyeron en los grupos tratados con Mel. Se descubrió que Mel tenía efectos protectores y terapéuticos. Según nuestros resultados, el efecto terapéutico de Mel en la lesión testicular aguda inducida por tioacetamida es mayor que su efecto protector.

Animals , Male , Rats , Testis/drug effects , Thioacetamide/toxicity , Melatonin/pharmacology , Antioxidants/pharmacology , Enzyme-Linked Immunosorbent Assay , Immunohistochemistry , Rats, Wistar , Heat-Shock Proteins/drug effects , Heat-Shock Proteins/metabolism , Melatonin/administration & dosage , Antioxidants/administration & dosage
Int. j. morphol ; 37(2): 522-532, June 2019. graf
Article in English | LILACS | ID: biblio-1002254


Amelogenin is one of the enamel matrices secreted by ameloblasts. A mutation of the amelogenin gene can cause hereditary dental enamel defects known as amelogenesis imperfecta (AI). Since lysosome-associated membrane protein-1 (LAMP-1), -3 (LAMP-3), and 78kDa glucose-related protein (Grp78) were identified as binding proteins of amelogenin, several studies have suggested the involvement of these binding proteins with the cell kinetics of ameloblasts in normal or abnormal conditions. The purpose of this study is to investigate the distribution of these amelogenin binding proteins in the ameloblast cell differentiation of mice with a point mutation of the amelogenin gene (Amelx*). The incisors of Amelx* mice had a white opaque color and the tooth surface was observed to be rough under a scanning electron microscope. Among the sequential ameloblast cell differentiation in the Amelx* mice, the shape of ameloblasts at the transition stage was irregular in comparison to those in wild-type (WT) mice. Immunostaining of Grp78 revealed that the whole cytoplasm of the transition stage ameloblasts was immunopositive for Grp78 antibody, while only the distal part of cell was positive in the WT mice. Furthermore, in the Amelx* mice, the cytoplasm of the transition stage ameloblasts was immunopositive for LAMP-1 and LAMP-3. These results suggest that Amelx* may cause the abnormal distribution of amelogenin binding proteins in the cytoplasm of ameloblasts.

La amelogenina es una de las matrices de esmalte secretadas por los ameloblastos. Una mutación del gen de amelogenina puede causar defectos hereditarios del esmalte dental conocidos como amelogénesis imperfecta (AI). Dado que la proteína de membrana asociada a lisosoma-1 (LAMP-1), -3 (LAMP-3) y la proteína relacionada con la glucosa de 78 kDa (Grp78) se identificaron como proteína de unión a amelogenina, varios estudios han sugerido la participación de estas proteínas con la cinética celular de los ameloblastos en condiciones normales o anormales. El objetivo del estudio fue investigar la distribución de LAMP-1, LAM-3 y Grp78 durante la diferenciación celular de ameloblastos de ratones con una mutación puntual del gen de amelogenina (Amelx*). Los incisivos de los ratones Amelx* presentaron un color blanco opaco y se observó en microscopio electrónico de barrido que la superficie del diente era áspera. La diferenciación celular secuencial y la forma de los ameloblastos en la etapa de transición en los ratones Amelx* fue irregular en comparación con los ratones silvestres (RS). La inmunotinción de Grp78 reveló que todo el citoplasma de los ameloblastos en etapa de transición fue inmunopositivo para el anticuerpo Grp78, mientras que solo la parte distal de la célula fue positiva en los ratones RS. Además, en ratones Amelx*, el citoplasma de los ameloblastos en etapa de transición fue inmunopositivo para LAMP-1 y LAMP-3. Estos resultados sugieren que Amelx* puede causar distribución anormal de proteínas de unión a amelogenina en el citoplasma de los ameloblastos.

Animals , Mice , Lysosome-Associated Membrane Glycoproteins/metabolism , Amelogenin/metabolism , Amelogenesis Imperfecta , Heat-Shock Proteins/metabolism , Microscopy, Electron, Scanning , Fluorescent Antibody Technique , Dental Enamel/pathology , Lysosomal-Associated Membrane Protein 1/metabolism , Amelogenin/genetics , Lysosomal-Associated Membrane Protein 3/metabolism , Incisor/pathology
Braz. j. med. biol. res ; 51(3): e7033, 2018. tab, graf
Article in English | LILACS | ID: biblio-889046


In the present study, we successfully demonstrated for the first time the existence of cardiac proteomic differences between non-selectively bred rats with distinct intrinsic exercise capacities. A proteomic approach based on two-dimensional gel electrophoresis coupled to mass spectrometry was used to study the left ventricle (LV) tissue proteome of rats with distinct intrinsic exercise capacity. Low running performance (LRP) and high running performance (HRP) rats were categorized by a treadmill exercise test, according to distance run to exhaustion. The running capacity of HRPs was 3.5-fold greater than LRPs. Protein profiling revealed 29 differences between HRP and LRP rats (15 proteins were identified). We detected alterations in components involved in metabolism, antioxidant and stress response, microfibrillar and cytoskeletal proteins. Contractile proteins were upregulated in the LVs of HRP rats (α-myosin heavy chain-6, myosin light chain-1 and creatine kinase), whereas the LVs of LRP rats exhibited upregulation in proteins associated with stress response (aldehyde dehydrogenase 2, α-crystallin B chain and HSPβ-2). In addition, the cytoskeletal proteins desmin and α-actin were upregulated in LRPs. Taken together, our results suggest that the increased contractile protein levels in HRP rats partly accounted for their improved exercise capacity, and that proteins considered risk factors to the development of cardiovascular disease were expressed in higher amounts in LRP animals.

Animals , Male , Rats , Physical Conditioning, Animal/physiology , Running/physiology , Proteins/metabolism , Heart Function Tests/methods , Myocardium/metabolism , Organ Size , Rats, Inbred Strains , Mass Spectrometry , Electrophoresis, Gel, Two-Dimensional , Proteins/isolation & purification , Contractile Proteins/metabolism , Cytoskeletal Proteins/metabolism , Proteomics , Desmin/metabolism , Heart Ventricles/metabolism , Heat-Shock Proteins/metabolism
Electron. j. biotechnol ; 29: 7-12, sept. 2017. ilus, graf, tab
Article in English | LILACS | ID: biblio-1016095


Background: DegP is a serine protease that specifically cleaves and refolds unfolding proteins in the periplasmic space of the cells. To date, there is no information regarding DegP from halophilic bacteria. Chromohalobacter salexigens BKL5 is a moderately halophilic bacterium that has the ability to grow in a media containing more than 15% salt. Therefore, the objectives of this work were to clone and overexpress DegP-encoding gene from C. salexigens BKL5 and characterize its biochemical properties. Results: DegP-encoding gene was overexpressed in Escherichia coli BL21(DE3) CodonPlus in an active form. SDS-PAGE analysis showed that the molecular weight of the recombinant DegP was 45 kDa. Size-exclusion chromatography analysis suggested that recombinant DegP was present in two multimeric states, hexameric and dodecameric, with molecular weights of 297.9 and 579.12 kDa, respectively. Both conformations were enzymatically active when casein was used as substrate for enzymatic assay. Circular dichroism analysis showed that recombinant DegP was composed of 0.21­0.29 helical content, which was comparable to the helical content in the crystal structure of E. coli DegP. The basic/acidic residue ratio of recombinant DegP was 0.56, which was slightly higher than that of DegP from extreme halophiles (average, 0.45) but significantly lower than that of DegP from nonhalophiles (average, 0.94). Conclusions: Recombinant DegP from C. salexigens BKL5 showed proteolytic activity when ß-casein was used as a substrate. In silico analysis indicated that recombinant DegP had characteristics similar to those of halophilic proteins depending on its amino acid composition.

Serine Endopeptidases/genetics , Periplasmic Proteins/genetics , Chromohalobacter/enzymology , Proteolysis , Heat-Shock Proteins/genetics , Recombinant Proteins , Serine Endopeptidases/metabolism , Caseins , Chromatography, Gel , Circular Dichroism , Cloning, Molecular , Periplasmic Proteins/metabolism , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Salinity , Chromohalobacter/genetics , Heat-Shock Proteins/metabolism , Molecular Weight
Acta cir. bras ; 31(3): 150-155, Mar. 2016. graf
Article in English | LILACS | ID: lil-777091


ABSTRACT PURPOSE : To investigate in the kidney the pathologic changes and expression of GRP78 and CHOP in the Kunming (KM) mice with combination of high-fat diet and streptozotocin-induced diabetes. METHODS : Sixty two male KM mice were randomly divided into a normal control (NC) group (n=20) and a high-fat diet (HFD) group (n=42). After a four-week dietary manipulation, the KM mice in the HFD group were injected intraperitoneally with streptozotocin to induce diabetes. After diabetic models were successfully established, the kidneys were excised and conserved for further test. RESULTS : No significant difference in the body weight was observed after the dietary manipulation (p=0.554). After the streptozotocin was injected, fasting blood glucose levels in the diabetes group (DM) were significantly higher than that in the NC group (p<0.0001). Glomerular atrophy observed under light microscope in the DM group was more serious compared with the NC group. The expression of GRP78 and CHOP in the kidneys of the mice in the DM group were higher compared with the NC group. CONCLUSION : Renal lesion occurs in the diabetic Kunming mice induced by combination of high-fat diet and low-dose streptozotocin, and endoplasmic reticulum stress and CHOP may contribute to the injury process.

Animals , Male , Mice , Diabetes Mellitus, Experimental/metabolism , Diabetic Nephropathies/metabolism , Diabetic Nephropathies/pathology , Endoplasmic Reticulum Stress/physiology , Diet, High-Fat , Blood Glucose/analysis , Body Weight/physiology , Random Allocation , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/pathology , Disease Models, Animal , Transcription Factor CHOP/metabolism , Unfolded Protein Response/physiology , Heat-Shock Proteins/metabolism , Kidney/metabolism , Kidney/pathology
Acta cir. bras ; 31(2): 143-149, Feb. 2016. graf
Article in English | LILACS | ID: lil-775565


PURPOSE: To investigate the anticancer activity of ellagic acid (EA) in U251 human glioblastoma cells and its possible molecular mechanism. METHODS: The cells were treated with EA at various concentrations for different time periods. Cell viability and cell proliferation were detected by cell counting kit-8(CCK-8) assay and live/dead assay respectively. Cell apoptosis were measured with Annexin V-FITC/PI double staining method by flow cytometry and Mitochondrial membrane potential assay separately. Cell cycle was measured with PI staining method by flow cytometry. The expressions of Bcl-2, Survivin, XIAP, Caspase-3, Bax, JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, DR4, DR5, CHOP and GRP78-related proteins were detected by western blot after EA treatment. RESULTS: Cell viability and proliferation of glioblastoma cells treated with EA were significantly lower than the control group. EA caused robust apoptosis of the glioblastoma cells compared to the control group. EA significantly decreased the proportion at G0/G1 phases of cell cycling accompanied by increased populations at S phase in U251 cell lines. And the expressions of anti-apoptotic proteins were dramatically down-regulated. CONCLUSION: Ellagic acid potentially up-regulated DR4, DR5 and MAP kinases (JNK, ERK1/2 and p38). EA also caused significant increase in the expressions of CHOP and GRP78. Our findings suggest that EA would be beneficial for the treatment of glioblastoma.

Humans , Apoptosis/drug effects , Glioblastoma/metabolism , Cell Proliferation/drug effects , Ellagic Acid/pharmacology , Cell Survival/drug effects , Apoptosis/physiology , MAP Kinase Signaling System/drug effects , Ellagic Acid/metabolism , Transcription Factor CHOP/drug effects , Transcription Factor CHOP/metabolism , Caspase 3/metabolism , Heat-Shock Proteins/drug effects , Heat-Shock Proteins/metabolism
Arq. bras. cardiol ; 105(1): 71-81, July 2015. tab, ilus
Article in English | LILACS | ID: lil-755009



Acute myocardial infarction is the leading cause of morbidity and mortality worldwide. Furthermore, research has shown that exercise, in addition to reducing cardiovascular risk factors, can also protect the heart against injury due to ischemia and reperfusion through a direct effect on the myocardium. However, the specific mechanism involved in exerciseinduced cardiac preconditioning is still under debate.


To perform a systematic review of the studies that have addressed the mechanisms by which aerobic exercise promotes direct cardioprotection against ischemia and reperfusion injury.


A search was conducted using MEDLINE, Literatura Latino-Americana e do Caribe de Informação em Ciências da Saúde, and Scientific Electronic Library Online databases. Data were extracted in a standardized manner by two independent researchers, who were responsible for assessing the methodological quality of the studies.


The search retrieved 78 studies; after evaluating the abstracts, 30 studies were excluded. The manuscripts of the remaining 48 studies were completely read and, of these, 20 were excluded. Finally, 28 studies were included in this systematic review.


On the basis of the selected studies, the following are potentially involved in the cardioprotective response to exercise: increased heat shock protein production, nitric oxide pathway involvement, increased cardiac antioxidant capacity, improvement in ATP-dependent potassium channel function, and opioid system activation. Despite all the previous investigations, further research is still necessary to obtain more consistent conclusions.



O infarto agudo do miocárdio é a principal causa de mortalidade e de morbidade na população mundial. Por outro lado, pesquisas já demonstraram que o exercício físico, além de reduzir os fatores de risco cardiovascular, também é capaz de promover cardioproteção contra lesões por isquemia e reperfusão, por meio de um efeito direto no miocárdio. No entanto, o mecanismo específico envolvido no pré-condicionamento cardíaco induzido pelo exercício ainda é alvo de discussão.


Realizar uma revisão sistemática acerca dos estudos que se debruçaram sobre os mecanismos pelos quais o exercício físico aeróbio promove cardioproteção direta contra lesões por isquemia e reperfusão.


Foi realizada uma pesquisa nas seguintes bases de dados: MEDLINE, LILACS e SciELO. Os dados foram extraídos de forma padronizada, por dois investigadores independentes, responsáveis pela avaliação da qualidade metodológica dos manuscritos.


A busca inicial resultou em 78 estudos, dos quais, após revisão dos resumos, 30 foram excluídos. Os 48 manuscritos restantes foram lidos na íntegra e, destes, 20 foram excluídos, restando 28 estudos incluídos nesta revisão sistemática.


Com base nos estudos selecionados, os seguintes mecanismos estão potencialmente envolvidos na resposta cardioprotetora do exercício: aumento na produção de proteínas de choque térmico; envolvimento da via do óxido nítrico; aumento na capacidade antioxidativa cardíaca; melhora na função dos canais de potássio dependentes de ATP; e ativação do sistema de opióides. Apesar de todo o investimento já realizado, ainda é necessário mais investimento em trabalhos futuros, para obtenção de conclusão mais consistente.


Humans , Exercise Therapy/methods , Exercise/physiology , Myocardial Infarction/metabolism , Myocardial Reperfusion Injury/metabolism , Antioxidants/metabolism , Heat-Shock Proteins/metabolism , KATP Channels/metabolism , Myocardial Infarction/physiopathology , Myocardial Infarction/prevention & control , Myocardial Reperfusion Injury/physiopathology , Myocardial Reperfusion Injury/prevention & control , Time Factors
Biol. Res ; 48: 1-5, 2015. graf
Article in English | LILACS | ID: biblio-950819


BACKGROUND: In the central nervous system, interleukin-10 (IL-10) provides trophic and survival effects directly on neurons, modulates neurite plasticity, and has a pivotal importance in the neuronal regeneration in neurodegenerative and neuroinflammatory conditions. This cytokine is primarily produced by glial cells and has beneficial effects on the neuronal viability. However, the mechanisms of IL-10-elicited neuroprotection are not clear. RESULTS: Membrane preparations, isolated from wild-type (Wt) and IL-10 knockout (KO) mice brain were used in this study. It has been shown that compared to wild-type mice, in IL-10 KO mice brain, the amount of immunoglobulin binding protein (BiP) is greatly increased, whereas the content of sigma receptor-1 (SigR1) is not changed significantly. Co-immunoprecipitation experiments have shown that the association of SigR1 with small GTPase Rac1 (Ras-related C3 botulinum toxin substrate 1), NR2B subunit of NMDA-receptor (NMDAR) and inositol-3-phosphate receptor (IP3R) is higher in the IL-10 KO mice brain than in the Wt mice brain. Besides, we have found that either glutamate or sigma ligands, separately or together, do not change glutamate-induced NADPH-oxidase (NOX) activity in Wt-type mice brain membrane preparations, whereas in IL-10 KO mice high concentration of glutamate markedly increases the NOX-dependent production of reactive oxygen species (ROS). Glutamate-dependent ROS production was decreased to the normal levels by the action of sigma-agonists. CONCLUSIONS: It has been concluded that IL-10 deprivation, at least in part, can lead to the induction of ER-stress, which causes BiP expression and SigR1 redistribution between components of endoplasmic reticulum (ER) and plasma membrane. Moreover, IL-10 deficiency can change the specific organization of NMDAR, increasing the surface expression of SigR1-sensitive NR2B-containing NMDAR. In these conditions, glutamate-dependent ROS production is greatly increased leading to the initiation of apoptosis. In this circumstances, sigma-ligands could play a preventive role against NMDA receptor-mediated excitotoxicity.

Animals , Male , Mice , Brain/metabolism , Interleukin-10/genetics , Receptors, sigma/metabolism , Glutamic Acid/metabolism , NADPH Oxidases/metabolism , Cell Membrane/metabolism , Receptors, sigma/classification , Receptors, sigma/agonists , Reactive Oxygen Species/analysis , Reactive Oxygen Species/metabolism , Receptors, N-Methyl-D-Aspartate/classification , Receptors, N-Methyl-D-Aspartate/metabolism , rac1 GTP-Binding Protein/metabolism , Immunoprecipitation , Endoplasmic Reticulum/metabolism , Inositol 1,4,5-Trisphosphate Receptors/metabolism , Gene Knockdown Techniques , Heat-Shock Proteins/metabolism , Mice, Inbred C57BL , Neurons/metabolism
Yonsei Medical Journal ; : 1497-1502, 2015.
Article in English | WPRIM | ID: wpr-177077


PURPOSE: Heat shock proteins (HSPs) are highly conserved molecular chaperones. There are various studies that assess the prognostic value of HSPs in patients with esophageal cancer, but the conclusion remains controversial. This is the first meta-analysis study aiming to summarize the evidence on the suitability of HSPs to predict patients' survival. MATERIALS AND METHODS: Searching PubMed, Web of science and Medline until May 31, 2014, data were compared for overall survival in patients with down-regulated HSPs level with those with up-regulated level. We conducted a meta-analysis of 9 studies (801 patients) that correlated HSPs levels with overall survival. Data were synthesized with hazard ratios (HRs). RESULTS: The estimated risk of death was 2.93-fold greater in HSP27 negative patients than HSP27 positive patients [95% confidence interval (CI), 1.12-7.62]. When limited to esophageal squamous cell carcinoma (ESCC), the risk of death in HSP27 negative patients seemed more significant (HR, 3.90; 95% CI, 2.35-6.49). Decreased expression of HSP70 was also associated with worse survival in esophageal cancer (HR, 2.83; 95% CI, 1.90-4.23) and, when limited to ESCC, HR was 3.21 (95% CI, 1.94-5.30). Data collected, however, were not sufficient to determine the prognostic value of HSP90 in patients with ESCC nor esophageal adenocarcinomas (EADC). CONCLUSION: In this meta-analysis, reduced HSP27 and HSP70 expressions were associated with poor survival in patients with esophageal cancer, especially esophageal squamous cell carcinoma.

Adenocarcinoma/diagnosis , Carcinoma, Squamous Cell/diagnosis , Esophageal Neoplasms/diagnosis , Gene Expression Regulation, Neoplastic , HSP27 Heat-Shock Proteins , HSP70 Heat-Shock Proteins , HSP90 Heat-Shock Proteins , Heat-Shock Proteins/metabolism , Humans , Male , Neoplasm Proteins , Prognosis , Survival , Treatment Outcome
Indian J Biochem Biophys ; 2013 Apr; 50(2): 126-138
Article in English | IMSEAR | ID: sea-147296


Abiotic stress causes abrupt increase in the expression of stress-associated proteins, which provide tolerance by modulating the defense mechanism of plants. Small heat shock proteins (sHSPs) and anti-oxidant enzymes are important for environmental stress tolerance of the plants. In this study, two full-length cDNAs encoding small heat shock protein (sHSP) and superoxide dismutase (SOD), designated as TasHSP and SODI were identified and characterized from C-306 (thermotolerant) and PBW343 (thermosusceptible) cultivars of wheat (Triticum aestivum L.). An alpha crystalline domain was observed in TasHSP and manganese/iron binding domain in case of SODI. Quantitative real-time PCR showed very high transcript level of TasHSP and SOD in C-306 compared to PBW343 at different stages of growth and against differential heat stress (HS). Under differential HS at milky-dough stage, the fold change in transcript of both TasHSP and SOD was observed maximum in C-306, compared to PBW343. Protein profiling and isoenzymes analysis showed the expression of several heat-stable proteins and prominent isoenzymes of SOD in C-306, compared to PBW343. Scanning electron microscopy (SEM) of starch granules showed globular, well-shaped and more numbers of endospermic cells in C-306, compared to defragmented, irregular shaped and shrunken granules in case of PBW343 under HS treatment (42°C for 2 h). Diurnal change in soluble starch synthase (SSS) activity showed an increase in the activity during afternoon (35°C), compared to morning (29°C) and evening (32°C) in both the cultivars. Under heat stress (42°C for 2 h), a drastic decrease in the SSS activity was observed, due to the thermal denaturation of the enzyme. Thermotolerance capacity analyzed using cell membrane stability (CMS) showed significantly higher CMS in case of C-306, compared to PBW343 at different stages of growth. Findings suggest that abundance of TasHSP and SODI during milky-dough stage plays a very important role in starch granule biosynthesis. The mechanism may be further exploited to develop tolerant wheat cultivar with high quality seeds.

Amino Acid Sequence , Base Sequence , Cell Membrane/metabolism , Cloning, Molecular , DNA, Complementary/metabolism , Gene Expression Profiling , Gene Expression Regulation, Plant , Heat-Shock Proteins/metabolism , Hot Temperature , Isoenzymes/metabolism , Microscopy, Electron, Scanning , Models, Biological , Molecular Sequence Data , Phylogeny , Real-Time Polymerase Chain Reaction/methods , Sequence Homology, Amino Acid , Starch/metabolism , Starch Synthase/metabolism , Superoxide Dismutase/metabolism , Triticum/metabolism
Article in English | WPRIM | ID: wpr-123287


Benzo(a)pyrene (BaP) is a polycyclic aromatic hydrocarbon (PAH) that is easily introduced to humans via consumption of grilled or smoked meat. BaP causes harmful oxidative effects on cell development, growth and survival through an increase in membrane lipid peroxidation, oxidative DNA damage and mutagenesis. Therefore, the present study was conducted to evaluate the synergistic effects of BaP on oxidative stress in hepatic tumors. In this study, we established a hepatic tumor model by injecting rat hepatoma N1-S1 cells into healthy rats. Changes in the abundance of heat shock proteins (HSPs), antioxidant enzymes and pro-inflammatory cytokines were then investigated by western blot analysis. In addition, we examined changes in oxidative stress levels. Injection of N1-S1 cells or concomitant injection of BaP and N1-S1 cells resulted in the formation of hepatic tumors at the injection site. Evaluation of rat plasma reveals that hepatic tumors induced by BaP and N1-S1 cells expresses higher levels of Hsp27, superoxide dismutase (SOD), and tumor necrosis factor-alpha (TNF-alpha) when compared to those induced by N1-S1 cells only. The collective results of this study suggest that BaP exerts synergistic effects on the expression of HSP, cytokines and antioxidant enzymes in hepatic tumors induced by rat hepatoma N1-S1 cells.

Animals , Antioxidants/metabolism , Benzo(a)pyrene/pharmacology , Carcinoma, Hepatocellular/metabolism , Cell Line, Tumor/drug effects , Cytokines/metabolism , Heat-Shock Proteins/metabolism , Humans , Liver Neoplasms/enzymology , Male , Neoplasms, Experimental/metabolism , Oxidative Stress/drug effects , Rats , Rats, Sprague-Dawley
Indian J Biochem Biophys ; 2009 Dec; 46(6): 482-490
Article in English | IMSEAR | ID: sea-135231


In congenital heart disease (CHD), mechanical wall stress by increased pulmonary artery pressure and pulmonary blood flow is believed to play a pivotal role in the pathogenesis of pulmonary plexogenic arteriopathy (PPA). The pathogenesis of this disease that involves significant pulmonary arterial remodelling, is, however, largely unknown. In the systemic circulation, upregulation of HSP-70 and HSP-27 in the arterial wall occurs in response to acute hypertension, whereas HSP-60 and increased titres of anti-HSP-60 antibodies are associated with atherosclerotic vessel disease. We looked for the involvement of HSPs in the stress response of pulmonary endothelial and vascular smooth muscle cells in different abnormal hemodynamic conditions in patients with CHDs. We analyzed the expression pattern of HSP-27, HSP-70 and HSP-60 in lung biopsies of 38 patients with CHD, using immunohistochemistry. These included 4 individuals with an essentially normal pulmonary circulation, who served as controls. Immunoreactivity against HSP-27 and also against HSP-70 was present in the pulmonary endothelium and vascular smooth muscle cells of patients and controls in a similar pattern. In contrast, expression of HSP-60 was absent in pulmonary arteries of both patients and controls. In patients with advanced PPA, cells within plexiform lesions showed strong staining for HSP-27 and HSP-70, but were again negative for HSP-60. The intensity of immunoreactivity against HSP-70 correlated inversely with medial thickness of pre-acinar arteries (r = -0.32; p = 0.04). Expression of HSP-27 and HSP-70 did not correlate with hemodynamic parameters, although immunoreactivity against HSP27 tended to be increased in cases with high pulmonary artery pressure (r = 0.37; p = 0.16) and was highest in patients with flow-associated pulmonary hypertension (p<0.01). HSP-27 and HSP-70, but not HSP-60 are engaged in the stress response of cells of small pulmonary arteries in pulmonary plexogenic arteriopathy. HSP-27 and HSP-70 are increasingly expressed in the advanced proliferative lesions of this disease.

Case-Control Studies , Child , Child, Preschool , Gene Expression Regulation , Heart Diseases/genetics , Heart Diseases/metabolism , Heat-Shock Proteins/metabolism , Hemodynamics , Humans , Immunohistochemistry , Infant , Infant, Newborn , Lung/blood supply , Protein Transport , Pulmonary Artery/physiopathology , Young Adult
Indian J Exp Biol ; 2008 May; 46(5): 273-309
Article in English | IMSEAR | ID: sea-58005


Convincing evidence supports a role for oxidative stress in the pathogenesis of many chronic diseases. The model includes the formation of radical oxygen species (ROS) and the misassembly and aggregation of proteins when three tiers of cellular defence are insufficient: (a) direct antioxidative systems, (b) molecular damage repairing systems, and (c) compensatory chaperone synthesis. The aim of the present overview is to introduce (a) the basics of free radical and antioxidant metabolism, (b) the role of the protein quality control system in protecting cells from free radical damage and its relation to chronic diseases, (c) the basics of the ultraweak luminescence as marker of the oxidant status of biological systems, and (d) the research in human photon emission as a non-invasive marker of oxidant status in relation to chronic diseases. In considering the role of free radicals in disease, both their generation and their control by the antioxidant system are part of the story. Excessive free radical production leads to the production of heat shock proteins and chaperone proteins as a second line of protection against damage. Chaperones at the molecular level facilitate stress regulation vis-à-vis protein quali y control mechanisms. The manifestation of misfolded proteins and aggregates is a hallmark of a range of neurodegenerative disorders including Alzheimer's disease, Parkinson's disease, amylotrophic lateral sclerosis, polyglutamine (polyQ) diseases, diabetes and many others. Each of these disorders exhibits aging-dependent onset and a progressive, usually fatal clinical course. The second part reviews the current status of human photon emission techniques and protocols for recording the human oxidative status. Sensitive photomultiplier tubes may provide a tool for non-invasive and continuous monitoring of oxidative metabolism. In that respect, recording ultraweak luminescence has been favored compared to other indirect assays. Several biological models have been used to illustrate the technique in cell cultures and organs in vivo. This initiated practical applications addressing specific human pathological issues. Systematic studies on human emission have presented information on: (a) procedures for reliable measurements, and spectral analysis, (b) anatomic intensity of emission and left-right symmetries, (c) biological rhythms in emission, (d) physical and psychological influences on emission, (e) novel physical characteristics of emission, and (f) the identification of ultraweak photon emission with the staging of ROS-related damage and disease. It is concluded that both patterns and physical properties of ultraweak photon emission hold considerable promise as measure for the oxidative status.

Antioxidants/metabolism , Biophysics/methods , Free Radicals , Gene Expression Regulation , Heat-Shock Proteins/metabolism , Hot Temperature , Humans , Hydrogen Peroxide/chemistry , Molecular Chaperones/metabolism , Neoplasms/metabolism , Oxidative Stress , Oxygen/chemistry , Photons , Reactive Oxygen Species , Reperfusion Injury
J Biosci ; 2007 Apr; 32(3): 489-99
Article in English | IMSEAR | ID: sea-110744


For many years,we and our collaborators have investigated the adaptive role of heat shock proteins in different animals,including the representatives of homothermic and poikilothermic species that inhabit regions with contrasting thermal conditions. Adaptive evolution of the response to hyperthermia has led to different results depending upon the species. The thermal threshold of induction of heat shock proteins in desert thermophylic species is, as a rule, higher than in the species from less extreme climates.In addition,thermoresistant poikilothermic species often exhibit a certain level of heat shock proteins in cells even at a physiologically normal temperature. Furthermore,there is often a positive correlation between the characteristic temperature of the ecological niche of a given species and the amount of Hsp70-like proteins in the cells at normal temperature. Although in most cases adaptation to hyperthermia occurs without changes in the number of heat shock genes, these genes can be amplified in some xeric species. It was shown that mobile genetic elements may play an important role in the evolution and fine-tuning of the heat shock response system,and can be used for direct introduction of mutations in the promoter regions of these genes.

Acclimatization/physiology , Animals , Desert Climate , Drosophila/genetics , Ecosystem , Biological Evolution , Heat-Shock Proteins/metabolism , Lizards/genetics , Moths/genetics , Promoter Regions, Genetic/genetics , Temperature
J Biosci ; 2007 Apr; 32(3): 511-5
Article in English | IMSEAR | ID: sea-110693


Environmental stress induces damage that activates an adaptive response in any organism. The cellular stress response is based on the induction of cytoprotective proteins,the so called stress or heat shock proteins. The stress response as well as stress proteins are ubiquitous,highly conserved mechanism, and genes, respectively, already present in prokaryotes. Chaperones protect the proteome against conformational damage, promoting the function of protein networks. Protein damage takes place during aging and in several degenerative diseases, and presents a threat to overload the cellular defense mechanisms. The preservation of a robust stress response and protein disposal is indispensable for health and longevity. This review summarizes the present knowledge of protein damage, turnover, and the stress response in aging and degenerative diseases.

Aging , Animals , Disease Susceptibility , Heat-Shock Proteins/metabolism , Humans , Signal Transduction , Stress, Physiological/metabolism
Rev. Fac. Cienc. Méd. (Córdoba) ; 63(3): 17-23, 2006. ilus
Article in Spanish | LILACS | ID: lil-474455


Introducción: la infección y la inflamación crónica han sido implicadas como agentes etiológicos para la ateraesclerasis (ATE). Varios estudios han relacionado a la infección por H. Pylori (HP) con la EC, especialmente con los linajes mas virulentos (linaje Cag A). Objetivo: demostrar la presencia del HP en placas de endarterectomías, utilizando una técnica Inmunohistoquimica (lHQ) específica que revela una reacción Ag-Ac mediante un cromógeno. Material y Métodos: se estudiaron 34 placas ATE de distintos territorios vasculares. Se fijaron en formol descalcificándolas en ácido fórmico según necesidad. Fueron incluidos en parafina, cortados y coloreados con H-E y técnicas de IHQ específicas para HP. Luego fueron desparafinados y tratados térmicamente con una solución de recuperación antigénica (lnmuno DNA Retriever with Citrate) utilizando olla a presión. La IHQ se efectuó con un sistema de alta sensibilidad Biotina-Estreptavidina-Peroxidasa-DAB). La observación morfológica evaluó células inflamatorias mononucleares y la identificación de la bacteria en la pared o la luz vascular. Resultados: de los 34 casos estudiados, en 14 se pudo identificar el bacilo en sus diferentes formas (41,17%), asociado a signos de inflamación crónica. Conclusión: el HP estuvo presente en un número sustancial de lesiones ATE y se asoció con inflamación. Estudios recientes sugieren que la presencia de Hp, demostrada por técnicas de IHQ, potenciaría los FR para ATE, induciendo una respuesta celular inflamatoria crónica por irritación persistente de la pared arterial.

Introduction: In general, infection and chronic inflammation have be en implied as etiologic agents for atherosclerosis and in particular coronary illness (CI). Several studies have correlated the infection of Helicobacter pylori with CI, especially with virulent strains (lineage Cag A). Objective: Demonstrate the immunohistochemical presence of H. Pylori in atherosclerotic plaques obtained from endarterectomy of different vascular regions. Material and methods: 34 atherosclerotic plaques of different vascular areas were studied, (25 men and 9 women). The tissues were fixed with 10% neutral buffered-formalin and decalcifying in formic acid 5% was used when necessary. The tissue sections were included in paraffin, cut and colored with H&E and subjected to Immunohistochemistry (lHC) of H.Pylori. Briefly, tissues were deparaffinized and thermally treated with a citrate-based solution of antigenic retrieval (lmmunoDNA Retriever with Citrate, BlO SB, Santa Barbara, CA) using a water bath at 95°C for 1 hour. The IHC was conducted using a high sensitivity BiotinStreptavidin-HRP-DAB IHC system (lmmunoDetector HRPIDAB, BlO SB). The microscopic observation evaluated the' presence of mononuclear inflammatory cells and the identification of the bacteria in the wall or the vascular lumen. Results: Of the 34 cases studied 14 were positive, where one could identify the bacillus in their different forms (41, 17%) associated with chronic inflammation.

Humans , Animals , Male , Female , Middle Aged , Atherosclerosis/microbiology , Helicobacter Infections/diagnosis , Helicobacter pylori/isolation & purification , Atherosclerosis/pathology , Endarterectomy , Heat-Shock Proteins/metabolism , Immunohistochemistry , Monocytes/metabolism
Article in English | WPRIM | ID: wpr-207077


Vimentin is an intermediate filament that regulates cell attachment and subcellular organization. In this study, vimentin filaments were morphologically altered, and its soluble subunits were rapidly reduced via cadmium chloride treatment. Cadmium chloride stimulated three major mitogen-activated protein kinases (MAPKs): extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38, and led apoptotic pathway via caspase-9 and caspase-3 activations. In order to determine whether MAPKs were involved in this cadmium-induced soluble vimentin disappearance, we applied MAPK- specific inhibitors (PD98059, SP600125, SB203580). These inhibitors did not abolish the cadmium-induced soluble vimentin disappearance. Caspase and proteosome degradation pathway were also not involved in soluble vimentin disappearance. When we observed vimentin levels in soluble and insoluble fractions, soluble vimentin subunits shifted to an insoluble fraction. As we discovered that heat- shock protein 27 (HSP27) was colocalized and physically associated with vimentin in unstressed cells, the roles of HSP27 with regard to vimentin were assessed. HSP27-overexpressing cells prevented morphological alterations of the vimentin filaments, as well as reductions of soluble vimentin, in the cadmium-treated cells. Moreover, HSP27 antisense oligonucleotide augmented these cadmium-induced changes in vimentin. These findings indicate that HSP27 prevents disruption of the vimentin intermediate filament networks and soluble vimentin disappearance, by virtue of its physical interaction with vimentin in cadmium-treated SK-N-SH cells.

Cadmium/pharmacology , Caspases/metabolism , Cell Line , Heat-Shock Proteins/metabolism , Humans , Mitogen-Activated Protein Kinases/metabolism , Protein Binding/drug effects , Protein Subunits/chemistry , Solubility/drug effects , Vimentin/chemistry
Article in English | WPRIM | ID: wpr-177643


During radiotherapy of cancer, neighboring normal cells may receive sub-lethal doses of radiation. To investigate whether such low levels of radiation modulate normal cell responses to death stimuli, primary cultured human fibroblasts were exposed to various doses of gamma-rays. Analysis of cell viability using an exclusion dye propidium iodide revealed that the irradiation up to 10 Gy killed the fibroblasts only to a minimal extent. In contrast, the cells efficiently lost their viability when exposed to 0.5-0.65 mM H2O2. This type of cell death was accompanied by JNK activation, and was reversed by the use of a JNK-specific inhibitor SP600125. Interestingly, H2O2 failed to kill the fibroblasts when these cells were pre-irradiated, 24 h before H2O2 treatment, with 0.25-0.5 Gy of gamma-rays. These cytoprotective doses of gamma-rays did not enhance cellular capacity to degrade H2O2, but elevated cellular levels of p21Cip/WAF1, a p53 target that can suppress H2O2-induced cell death by blocking JNK activation. Consistently, H2O2-induced JNK activation was dramatically suppressed in the pre-irradiated cells. The overall data suggests that ionizing radiation can impart normal fibroblasts with a survival advantage against oxidative stress by blocking the process leading to JNK activation.

Antioxidants/pharmacology , Cell Death , Cells, Cultured , Enzyme Activation/radiation effects , Fibroblasts/enzymology , Gamma Rays , Heat-Shock Proteins/metabolism , Humans , JNK Mitogen-Activated Protein Kinases/antagonists & inhibitors , Oxidative Stress/radiation effects , Water/pharmacology