Cathepsin D repairing role in photodamaged skin barrier.

UNLABELLED: BACKGROUNDS/OBJECTIVES: Cathepsin D plays an important part in maintaining a normal skin barrier. Our previous study found that cathepsin D decreased in chronic photodamaged skin. This study investigated the cathepsin D content change in the stratum corneum (SC) and the repairing role of cathepsin D in chronic photodamaged skin barrier via the application of cathepsin D gel. METHODS: Cathepsin D gel (0.001%) was applied to chronic photodamaged (sun-exposed forearm) human skin on identical sites (1 cm(2)/area) twice daily for 2 weeks. At 30 min and at 1, 3, 7, and 14 days, skin hydration and transepidermal water loss (TEWL) average values were detected via noninvasive skin detection equipment. Cathepsin D and transglutaminase (TGase)-1 in the skin sublayers were separated and detected via tape stripping, ELISA and Western blot. RESULTS: After 2 weeks of cathepsin D gel application, the skin moisture value increased from 86.8 ± 1.2 to 95.2 ± 2.7 (p < 0.05), while TEWL decreased from 17.88 ± 1.87 to 11.58 ± 2.14 (p < 0.05). Cathepsin D protein was detected in the upper epidermis (12.6 ± 2.6 ng/cm(2)), mid-epidermis (8.4 ± 0.8 ng/cm(2)) and deep epidermis (16.2 ± 2.6 ng/cm(2)) in the cathepsin D gel group compared to the control group (2.2 ± 0.7, 3.0 ± 1.1 and 3.85 ± 1.4 ng/cm(2), respectively; p < 0.05). TGase-1 enzyme expression was upregulated 2.54 ± 0.19 times in the matrix gel-treated skin. CONCLUSIONS: These data suggest that cathepsin D gel could increase the SC cathepsin D content and repair the epidermal barrier in chronic photodamaged skin. The mechanism might be related to increasing TGase-1 expression and activity.

Expresión de la catepsina D en epitelio escamoso cervical y neoplasia cervical intraepitelial / Cathepsin D expression in cervical squamous epithelia and cervical intraepithelial neoplasia

Objetivo: Determinar las positividades mediante técnicas inmunohistoquímicas de la catepsina D (CD) en nuestras muestras así como la intensidad y localización en el espesor del epitelio de los casos positivos para este marcador. Material y métodos: Nuestro estudio está formado por 113 muestras de tejido cervical divididas en 4 grupos: 30 casos de epitelio escamoso cervical normal (CC), 31 de neoplasia cervical intraepitelial grado 1 (CIN1), 27 de neoplasia cervical intraepitelial grado 2 (CIN2) y 25 de neoplasia cervical intraepitelial grado 3 (CIN3). Se obtienen cortes de tejido, tras su fijación e inclusión en parafina, en los que se aplica una técnica inmunohistoquímica específica para la detección de la CD. Resultados: La distribución de las positividades de este marcador fueron 0 por ciento para el CC, 48 por ciento para el CIN1, 33 por ciento para el CIN2 y 44 por ciento para el CIN3, con p= 0,00001 en la relación CC-CIN1, p = 0,0005 en la relación CC-CIN2 y p = 0,00005 en la relación CCCIN3 (el nivel de significación estadística aplicado ha sido del 5 por ciento). La diferencia en la distribución de las intensidades fue significativa en el grupo de tinción intensa: 6,6 por ciento para CIN1, 0 por ciento para CIN2 y 45 por ciento para CIN3 (p = 0,0476). Conclusión: La expresión de la CD en un alto porcentaje de CIN1 puede atribuirse a su función estimuladora de la mitosis, así como el aumento de intensidad en los CIN3 podría indicar la adquisición temprana de alteraciones propias del fenotipo metastásico. Son necesarios estudios posteriores para sentar como base de la oncogénesis cervical nuestras conclusiones (AU)

La afectación ganglionar en los carcinomas escamosos de pulmón.Interés de la proteína oncogénica erbB2 y el Cyfra 21.1 / Lymph nodes involvement from lung squamous carcinomas. Interest of erbB2 oncogenic protein and Cyfra 21.1

No disponible

Microinjection of cathepsin d induces caspase-dependent apoptosis in fibroblasts.

Recent reports have indicated that enzymes such as cathepsins D and B are translocated from lysosomal compartments to the cytosol early during apoptosis. We have previously noted that a translocation of cathepsins D and B occur before cytochrome c release and caspase activation in cardiomyocytes and human fibroblasts during oxidative stress-induced apoptosis. In the present report, we use a microinjection technique to investigate if cytosolic location of the cathepsins D and B are important for induction of apoptosis. We found that microinjection of cathepsin D into the cytosol of human fibroblasts caused apoptosis, which was detected as changes in distribution of cytochrome c, cell shrinkage, activation of caspases, chromatin condensation, and formation of pycnotic nuclei. No apoptosis was, however, induced by microinjection of cathepsin B. Moreover, apoptosis was prevented in fibroblasts pretreated with a caspase-3-like inhibitor, and also when microinjected with cathepsin D mixed with the cathepsin D inhibitor, pepstatin A. These results show that cytosolic cathepsin D can act as a proapoptotic mediator upstream of cytochrome c release and caspase activation in human fibroblasts.

Human neutrophil azurocidin synergizes with leukocyte elastase and cathepsin G in the killing of Capnocytophaga sputigena.

Azurocidin was purified in the presence of phenylmethylsulfonyl fluoride. Electrophoresis revealed at least seven species which exhibited N-terminal sequences consistent with azurocidin. Azurocidin exhibited no bactericidal activity against Capnocytophaga sputigena or other oral bacteria but synergized the bactericidal activity of enzymatically active elastase. Azurocidin also interacted synergistically with cathepsin G.

A method for [3H]mannose labeling of Asn-linked oligosaccharides on recombinant glycoproteins synthesized in Xenopus oocytes.

We have developed an efficient method for labeling the Asn-linked oligosaccharides of recombinant glycoproteins synthesized in Xenopus laevis oocytes. By coinjecting GDP-[3,4-(3)H]mannose with mRNA for human cathepsin D, it was possible to incorporate as much as 1800 cpm per oocyte into each of the two Asn-linked oligosaccharides of this glycoprotein. Overall, about 50% of the microinjected GDP-[3,4-(3)H]mannose was incorporated into Asn-linked oligosaccharides, a 10-fold greater value than that obtained when [2-(3)H]mannose was microinjected. Less than 10% of the injected GDP-[3,4-(3)H]mannose was metabolized to water or converted to amino acids. This technique should facilitate studies of Asn-linked oligosaccharide biosynthesis, processing, and structure in recombinant proteins synthesized in Xenopus oocytes.

[Pro-cathepsin D can activate in vitro pro-cathepsin B secreted by ovarian cancers]. / La pro-cathepsine D peut activer in vitro la pro-cathepsine B sécrétée par les cancers ovariens.

A precursor form of cathepsin B (Mr 45-47 kd) was purified from ascitic fluids of patients with ovarian adenocarcinomas. Following pepsin activation, this precursor produced a 33 kd cathepsin B-like proteinase closely related to lysosomal cathepsin B. A similar activation was found using the 52 kd pro-cathepsin D secreted by the MCF7 human breast cancer cells. This activation was a time, dose and pH dependent process. These results suggest that the 52 kd pro-cathepsin D may be involved in the early steps of the "metastatic cascade", activating pro-cathepsin B in an acidic environment.