Discordance in cathepsin B and cystatin C expressions in bronchoalveolar fluids between murine bleomycin-induced fibrosis and human idiopathic fibrosis

International audienceAbstractThe activity of cysteine cathepsin B increased markedly in lung homogenates and in bronchoalveolar lavage fluids (BALF) of the mouse model of bleomycin-induced lung fibrosis after 14 days of challenge. In contrast the level of the cysteine cathepsin inhibitor cystatin C was unaffected in BALF of wild-type and cathepsin B-deficient mice. Therefore, murine cystatin C is not a reliable marker of fibrosis during bleomycin-induced lung fibrosis. Current data are in sharp contrast to previous analysis carried on human BALF from patients with idiopathic pulmonary fibrosis, for which the level of cathepsin B remained unchanged while cystatin C was significantly increased

Удосконалення темперуючої машини для кондитерських мас

This paper reports the improved model of a tempering machine for heating the formulation mixture of marshmallow, characterized by heat supply to the working tank through the replacement of a steam jacket with heating by a film resistive electric heater of radiative type (FREhRT). The surface of the heat exchange of the device was increased by heating the stirrer with FREhRT; secondary energy (30....85 °C) was used by converting it by Peltier elements for the autonomous operation of superchargers for cooling the engine compartment. The proposed solution will lead to an increase in the efficiency of the device, which is explained by a decrease in its specific metal consumption through the use of FREhRT. A reduction in the duration of heating (75 °C) a marshmallow formulation mixture was experimentally established: in the examined model, 530 s, compared with the analog, 645 s. That confirmed the reduction in heating time to the set temperature by 21.7 % compared to the MT-250 basic design. The calculations have established a decrease, by 13 %, in the specific energy consumption for heating the volume of a unit of product when using the improved structure, 205.7 kJ/kg, when using the basic one ‒ 232.1 kJ/kg. The increase in the efficiency of the proposed structure is explained by a decrease in the specific metal consumption of the device from 474 kg/m2 in the base apparatus to 273 kg/m2 in the improved one. The study results confirm the increase in the resource efficiency of the improved tempering machine, which is achieved by eliminating the steam jacket; increasing the heat exchange surface by heating the stirrer. The heat transfer by FREhRT simplifies the operational performance of the temperature stabilization system in a working tank. The reported results could prove useful when designing thermal devices with electric heat supply under the conditions of using secondary energy, which is relevant for ensuring resource efficiencyВдосконалено модель темперуючої машини для нагрівання рецептурної суміші зефіру, яка відрізняється теплопідведенням робочої ємності шляхом заміни парової сорочки на нагрів плівковкоподібним резистивним електронагрівачем випромінювального типу (ПпРЕнВт). Збільшено поверхню теплообміну апарата за рахунок обігріву мішалки ПпРЕнВт та використано вторинну енергію (30….85 °С) за рахунок перетворення її елементами Пельтьє для автономної роботи нагнітачів по охолодженню моторного відсіку. Запропоноване рішення призведе до підвищення ефективності апарата, що пояснюється зменшенням його питомої металоємності за рахунок використання ПпРЕнВт. Експериментально встановлено зменшення тривалості нагрівання (75 °С) рецептурної суміші зефіру: дослідної моделі – 530 с, в порівнянні з аналогом – 645 с. Тим самим підтверджено скорочення часу нагріву до заданої температури на 21,7 % порівняно з базової конструкцією МТ-250. Розрахунковими дослідженнями встановлено зменшення на 13 % питомих витрат енергії на нагрівання об’єму одиниці продукту у вдосконаленої конструкції – 205,7 кДж/кг, для базової – 232,1 кДж/кг. Підвищення ефективності запропонованої конструкції пояснюється зменшенням питомої металоємності апарата з 474 кг/м2 базового апарату до 273 кг/м2 в удосконаленому. Результати досліджень підтверджують підвищення ресурсоефктивності удосконаленої темперуючої машини, що досягається: усуненням парової сорочки; збільшенням поверхні теплообміну обігріванням мішалки. Тепловідведення ПпРЕнВт спрошує експлуатаційні показники системи стабілізації температури у робочої ємності. Результати досліджень будуть корисні під час проектування теплових апаратів з електричним теплопідведенням в умовах використання вторинної енергії, що є актуальним при забезпеченні ресурсоефктивност

Roles of cysteine cathepsins and their naturals inhibitors, cystatins in lung fibrosis

Lors de la fibrose pulmonaire idiopathique (FPI), la différenciation fibroblastique s’accompagne d’une accumulation excessive des composants de la matrice extracellulaire ainsi qu’à un dérèglement de la balance protéases / antiprotéases. Nous avons étudié le rôle des cathepsines à cystéine dans la myofibrogenèse et leur contribution potentielle à la physiopathologie de la fibrose pulmonaire chez l’Homme. Pour cela, le profil d’expression des cathepsines ainsi que de leurs inhibiteurs naturels a été évalué dans un modèle cellulaire expérimental, puis dans des myofibroblastes primaires et enfin dans des liquides de lavage broncho-Alvéolaires (LBA) de patients atteints de FPI. Nos résultats montrent que lors de la FPI la cystatine C (inhibiteur naturel des protéases à cystéine) régule les activités protéolytiques des cathepsines extracellulaires et pourrait ainsi contribuer à l’accumulation de collagènes. Elle serait un biomarqueur potentiel de la FPI. D’autre part la cathepsine B participe à la différentiation fibroblastique et son inhibition retarde la myofibrogenèse.During idiopathic pulmonary fibrosis (IPF), fibroblast differentiation is accompanied by an excessive accumulation of extracellular matrix components as well as an imbalance between proteases and theirs inhibitors. We evaluated the role of human cysteine cathepsins in myofibrogenesis and their potential contribution to the pathogenesis of IPF. Expression of cathepsins and their natural inhibitors have been studied in an experimental cell model, but also in primary myofibroblasts and in bronchoalveolar lavage fluids (BALF) of patients suffering from IPF. Our results show that cystatin C (a natural inhibitor of cysteine proteases) regulates the extracellular proteolytic activities of cathepsins and could contribute to the accumulation of collagens. Cystatin C could also be a potential biomarker of IPF. On the other hand, cathepsin B participates in fibroblast differentiation and its inhibition delays myofibrogenesis

Cystatin M/E (Cystatin 6): A Janus-Faced Cysteine Protease Inhibitor with Both Tumor-Suppressing and Tumor-Promoting Functions

International audienceAlongside its contribution in maintaining skin homeostasis and its probable involvement in fetal and placental development, cystatin M/E (also known as cystatin 6) was first described as a tumor suppressor of breast cancer. This review aims to provide an update on cystatin M/E with particular attention paid to its role during tumorigenesis. Cystatin M/E, which is related to type 2 cystatins, displays the unique property of being a dual tight-binding inhibitor of both legumain (also known as asparagine endopeptidase) and cysteine cathepsins L, V and B, while its expression level is epigenetically regulated via the methylation of the CST6 promoter region. The tumor-suppressing role of cystatin M/E was further reported in melanoma, cervical, brain, prostate, gastric and renal cancers, and cystatin M/E was proposed as a biomarker of prognostic significance. Contrariwise, cystatin M/E could have an antagonistic function, acting as a tumor promoter (e.g., oral, pancreatic cancer, thyroid and hepatocellular carcinoma). Taking into account these apparently divergent functions, there is an urgent need to decipher the molecular and cellular regulatory mechanisms of the expression and activity of cystatin M/E associated with the safeguarding homeostasis of the proteolytic balance as well as its imbalance in cancer

Impact of cancer associated fibroblasts-derived cathepsin B on breast cancer progression

Tumors arise in a complex ecosystem gover ned by interactions established between cancer cells and the microenvironment. This one is constituted on one side by a multitude of different non-cancerous cell types (e.g.: stromal and immune cells) and on the other side by components of the extracellular matrix. During cancerogenesis fibroblasts are activated and differenciated into cancer associated fibroblasts (CAFs). Nevertheless, the precise functions of CAFs in cancer progression are not fully understood. Among proteases implicated in both ECM remodeling and cancer progression, cathepsin B (Ctsb), a lysosomial cystein protease, has been detected in cancer cells and in tumor-associated macrophages. Ctsb expression is associated with a poor prognosis in breast cancer patients. However, the contribution of CAF-derived Ctsb to tumor progression is unknown. By using the MMTV-PyMT mouse model of breast cancer, our preliminary data reveal that CAFs express Ctsb at higher levels than cancer cells. Our data show that Ctsb deficiency impairs the capacity of CAFs to interact with collagen fibers and strongly diminishes the migration of CAFs in a 3D spheroid model. Further more, we demonstrate that the invasion of Ctsb-/- CAFs is restored upon addition of conditioned medium collected from WT CAFs but Further more, the invasion of Ctsb-/- CAFs is restored upon addition of conditioned medium collected from wild-type (WT) CAFs but also by durotaxis or by deletion of Cathepsin Z (Ctsz). Collectively these data suggest that Ctsb represents a key regulator of the complex cross-talk established between CAFs, the ECM and cancer cells

Impact of cancer associated fibroblasts-derived cathepsin B on breast cancer progression

Tumors arise in a complex ecosystem gover ned by interactions established between cancer cells and the microenvironment. This one is constituted on one side by a multitude of different non-cancerous cell types (e.g.: stromal and immune cells) and on the other side by components of the extracellular matrix. During cancerogenesis fibroblasts are activated and differenciated into cancer associated fibroblasts (CAFs). Nevertheless, the precise functions of CAFs in cancer progression are not fully understood. Among proteases implicated in both ECM remodeling and cancer progression, cathepsin B (Ctsb), a lysosomial cystein protease, has been detected in cancer cells and in tumor-associated macrophages. Ctsb expression is associated with a poor prognosis in breast cancer patients. However, the contribution of CAF-derived Ctsb to tumor progression is unknown. By using the MMTV-PyMT mouse model of breast cancer, our preliminary data reveal that CAFs express Ctsb at higher levels than cancer cells. Our data show that Ctsb deficiency impairs the capacity of CAFs to interact with collagen fibers and strongly diminishes the migration of CAFs in a 3D spheroid model. Further more, we demonstrate that the invasion of Ctsb-/- CAFs is restored upon addition of conditioned medium collected from WT CAFs but Further more, the invasion of Ctsb-/- CAFs is restored upon addition of conditioned medium collected from wild-type (WT) CAFs but also by durotaxis or by deletion of Cathepsin Z (Ctsz). Collectively these data suggest that Ctsb represents a key regulator of the complex cross-talk established between CAFs, the ECM and cancer cells

Regulation of TGF-β1-Driven Differentiation of Human Lung Fibroblasts: Emerging Roles of Cathepsin B and Cystatin C

Lung matrix homeostasis partly depends on the fine regulation of proteolytic activities. We examined the expression of human cysteine cathepsins (Cats) and their relative contribution to TGF-β1-induced fibroblast differentiation into myofibroblasts. Assays were conducted using both primary fibroblasts obtained from patients with idiopathic pulmonary fibrosis and human lung CCD-19Lu fibroblasts. Pharmacological inhibition and genetic silencing of Cat B diminished α-smooth muscle actin expression, delayed fibroblast differentiation, and led to an accumulation of intracellular 50-kDa TGF-β1. Moreover, the addition of Cat B generated a 25-kDa mature form of TGF-β1 in Cat B siRNA-pretreated lysates. Inhibition of Cat B decreased Smad 2/3 phosphorylation but had no effect on p38 MAPK and JNK phosphorylation, indicating that Cat B mostly disturbs TGF-β1-driven canonical Smad signaling pathway. Although mRNA expression of cystatin C was stable, its secretion, which was inhibited by brefeldin A, increased during TGF-β1-induced differentiation of idiopathic pulmonary fibrosis and CCD-19Lu fibroblasts. In addition, cystatin C participated in the control of extracellular Cats, because its gene silencing restored their proteolytic activities. These data support the notion that Cat B participates in lung myofibrogenesis as suggested for stellate cells during liver fibrosis. Moreover, we propose that TGF-β1 promotes fibrosis by driving the effective cystatin C-dependent inhibition of extracellular matrix-degrading Cats

Curcumin inhibits the TGF-β1-dependent differentiation of lung fibroblasts via PPARγ-driven upregulation of cathepsins B and L

Abstract Pulmonary fibrosis is a progressive disease characterized by a widespread accumulation of myofibroblasts and extracellular matrix components. Growing evidences support that cysteine cathepsins, embracing cathepsin B (CatB) that affects TGF-β1-driven Smad pathway, along with their extracellular inhibitor cystatin C, participate in myofibrogenesis. Here we established that curcumin, a potent antifibrotic drug used in traditional Asian medicine, impaired the expression of both α-smooth muscle actin and mature TGF-β1 and inhibited the differentiation of human lung fibroblasts (CCD-19Lu cells). Curcumin induced a compelling upregulation of CatB and CatL. Conversely cystatin C was downregulated, which allowed the recovery of the peptidase activity of secreted cathepsins and the restoration of the proteolytic balance. Consistently, the amount of both insoluble and soluble type I collagen decreased, reaching levels similar to those observed for undifferentiated fibroblasts. The signaling pathways activated by curcumin were further examined. Curcumin triggered the expression of nuclear peroxisome proliferator-activated receptor γ (PPARγ). Contrariwise PPARγ inhibition, either by an antagonist (2-chloro-5-nitro-N-4-pyridinyl-benzamide) or by RNA silencing, restored TGF-β1-driven differentiation of curcumin-treated CCD-19Lu cells. PPARγ response element (PPRE)-like sequences were identified in the promoter regions of both CatB and CatL. Finally, we established that the transcriptional induction of CatB and CatL depends on the binding of PPARγ to PPRE sequences as a PPARγ/Retinoid X Receptor-α heterodimer