Myosin motors and not actin comets are mediators of the actin-based Golgi-to-endoplasmic reticulum protein transport

We have previously reported that actin filaments are involved in protein transport from the Golgi complex to the endoplasmic reticulum. Herein, we examined whether myosin motors or actin comets mediate this transport. To address this issue we have used, on one hand, a combination of specific inhibitors such as 2,3-butanedione monoxime (BDM) and 1-[5-isoquinoline sulfonyl]-2-methyl piperazine (ML7), which inhibit myosin and the phosphorylation of myosin II by the myosin light chain kinase, respectively; and a mutant of the nonmuscle myosin II regulatory light chain, which cannot be phosphorylated (MRLC2AA). On the other hand, actin comet tails were induced by the overexpression of phosphatidylinositol phosphate 5-kinase. Cells treated with BDM/ML7 or those that express the MRLC2AA mutant revealed a significant reduction in the brefeldin A (BFA)-induced fusion of Golgi enzymes with the endoplasmic reticulum (ER). This delay was not caused by an alteration in the formation of the BFA-induced tubules from the Golgi complex. In addition, the Shiga toxin fragment B transport from the Golgi complex to the ER was also altered. This impairment in the retrograde protein transport was not due to depletion of intracellular calcium stores or to the activation of Rho kinase. Neither the reassembly of the Golgi complex after BFA removal nor VSV-G transport from ER to the Golgi was altered in cells treated with BDM/ML7 or expressing MRLC2AA. Finally, transport carriers containing Shiga toxin did not move into the cytosol at the tips of comet tails of polymerizing actin. Collectively, the results indicate that 1) myosin motors move to transport carriers from the Golgi complex to the ER along actin filaments; 2) nonmuscle myosin II mediates in this process; and 3) actin comets are not involved in retrograde transport.

Decoloración de tintes por lacasa

Informe de investigación realizado a partir de una estancia en el Instituto de Biotecnología Medioambiental de la Universidad Tecnológica de Graz, Austria, entre Julio y Agosto de 2006. Se ha estudiado la decoloración de varios tintes sintéticos de estructuras químicas diferentes (Rojo Congo, Azul de Naftol, Indigo Carmín, Lanaset Gris, Azul de Nilo) por la enzima lacasa inmovilizada. La inmovilización de la enzima lacasa se llevó a cabo sobre esferas de alúmina (Al2O3) de 3 mm de diámetro debido a la resistecia mecánica de este material. La lacasa y la proteína inmovilizada se determinaron como la diferencia entre las concentraciones iniciales y residuales (obtenidas en los lavados). El porcentaje de lacasa inmovilizada fue del 68% y la cantidad de proteína inmovilizada por gramo de soporte de 5,6 mg. La enzima lacasa inmovilizada fue capaz de decolorar tintes de diferente estructura sin la necesidad de añadir mediadores redox, lo cual la hace una enzima muy adecuada para su aplicación en la decoloración de efluentes procedentes de la industria textil. De todas formas, son necesarios más estudios para optimizar la técnica de inmovilización así como el proceso de decoloración. Es interesante destacar que aunque hay muchos estudios basados en la oxidación de tintes textiles por lacasa la mayoría utilizan la enzima libre y hay muy poca información disponible sobre la aplicación de lacasas inmovilizadas, por lo que el presente estudio constituye una aportación muy interesante y novedosa.

És el conferencing una eina útil per als programes de mediació en l’àmbit penal del Departament de Justícia?

L'estudi té per objectiu valorar la conveniència d'implementar la pràctica del conferencing en els programes de mediació i reparació penal, tant en la jurisdicció penal juvenil com en la jurisdicció ordinària. La recerca inclou dos estudis: el primer és una anàlisi dels programes i models de conferencing desenvolupats en 4 països: Nova Zelanda, Austràlia, Irlanda del Nord i Bèlgica; el segon recull les percepcions dels mediadors de l'àmbit penal a Catalunya pel que fa a l'assoliment dels objectius dels programes, la inclusió espontània de més persones en el procés de mediació, el coneixement sobre el conferencing, l'opinió en relació amb la implementació d'aquesta pràctica, les necessitats formatives i els interessos de futur. La metodologia de la recerca combina aproximacions qualitatives i quantitatives, i inclou l'anàlisi de la documentació dels programes dels països estudiats, la creació, validació i passació a càrrec d’experts d'un qüestionari, i la realització de dos grups de discussió i d’una entrevista als professionals dels programes de mediació penal. Els mediadors consultats consideren que el conferencing és una eina útil per als programes de mediació en l'àmbit penal. L'estudi dels models implementats en d'altres països aporta elements útils per a l'adaptació del conferencing a la realitat i les especificitats de Catalunya. Així, es concreten aspectes que cal tenir en compte per a la definició d'un programa pilot.

¿Es el conferencing una herramienta útil para los programas de mediación en el ámbito penal del Departamento de Justicia?

El estudio analiza la práctica del conferencing en 4 países (Nueva Zelanda, Australia, Irlanda del Norte y Bélgica), y efectúa un estudio empírico que recoge las percepciones de los mediadores sobre si esta práctica restaurativa puede ser una herramienta útil para los programas de mediación en el ámbito penal.

Lactic acid bacteria from fresh fruit and vegetables as biocontrol agents of phytopathogenic bacteria and fungi

This study evaluated the efficacy of lactic acid bacteria (LAB) isolated from fresh fruits and vegetables as biocontrol agents against the phytopathogenic and spoilage bacteria and fungi, Xanthomonas campestris, Erwinia carotovora, Penicillium expansum, Monilinia laxa, and Botrytis cinerea. The antagonistic activity of 496 LAB strains was tested in vitro and all tested microorganisms except P. expansum were inhibited by at least one isolate. The 496 isolates were also analyzed for the inhibition of P. expansum infection in wounds of Golden Delicious apples. Four strains (TC97, AC318, TM319, and FF441) reduced the fungal rot diameter of the apples by 20%; only Weissella cibaria strain TM128 decreased infection levels by 50%. Cell-free supernatants of selected antagonistic bacteria were studied to determine the nature of the antimicrobial compounds produced. Organic acids were the preferred mediators of inhibition but hydrogen peroxide was also detected when strains BC48, TM128, PM141 and FF441 were tested against E. carotovora. While previous reports of antifungal activity by LAB are scarce, our results support the potential of LAB as biocontrol agents against postharvest rot. [Int Microbiol 2008; 11(4):231-236]

Tumor necrosis factor-alpha mediates changes in tissue protein turnover in a rat cancer cachexia model.

Rats bearing the Yoshida AH-130 ascites hepatoma showed enhanced fractional rates of protein degradation in gastrocnemius muscle, heart, and liver, while fractional synthesis rates were similar to those in non-tumor bearing rats. This hypercatabolic pattern was associated with marked perturbations of the hormonal homeostasis and presence of tumor necrosis factor in the circulation. The daily administration of a goat anti-murine TNF IgG to tumor-bearing rats decreased protein degradation rates in skeletal muscle, heart, and liver as compared with tumor-bearing rats receiving a nonimmune goat IgG. The anti-TNF treatment was also effective in attenuating early perturbations in insulin and corticosterone homeostasis. Although these results suggest that tumor necrosis factor plays a significant role in mediating the changes in protein turnover and hormone levels elicited by tumor growth, the inability of such treatment to prevent a reduction in body weight implies that other mediators or tumor-related events were also involved.

Dexamethasone inhibition of leucocyte adhesion to rat mesenteric postcapillary venules: role of intercellular adhesion molecule 1 and KC

BACKGROUND A previous study showed that the glucocorticoid dexamethasone, at doses of 100 ¿g/kg and above, inhibited leucocyte adhesion to rat mesenteric postcapillary venules activated with interleukin 1ß (IL-1ß), as assessed by videomicroscopy. AIMS To identify whether the adhesion molecule, intercellular adhesion molecule 1 (ICAM-1), or the chemokine KC could be targeted by the steroid to mediate its antiadhesive effect. METHODS Rat mesenteries were treated with IL-1ß (20 ng intraperitoneally) and the extent of leucocyte adhesion measured at two and four hours using intravital microscopy. Rats were treated with dexamethasone, and passively immunised against ICAM-1 or KC. Endogenous expression of these two mediators was validated by immunohistochemistry, ELISA, and the injection of specific radiolabelled antibodies. RESULTS Dexamethasone greatly reduced IL-1ß induced leucocyte adhesion, endothelial expression of ICAM-1 in the postcapillary venule, and release of the mast cell derived chemokine KC. Injection of specific antibodies to the latter mediators was also extremely effective in downregulating (>80%) IL-1ß induced leucocyte adhesion. CONCLUSIONS Induction by IL-1ß of endogenous ICAM-1 and KC contributes to leucocyte adhesion to inflamed mesenteric vessels. Without excluding other possible mediators, these data clearly show that dexamethasone interferes with ICAM-1 expression and KC release from mast cells, resulting in suppression of leucocyte accumulation in the bowel wall, which is a prominent feature of several gastrointestinal pathologies.

Insulin dysfunction and allostic load in bipolar disorder

Bipolar disorder (BD) is associated with substantial morbidity, as well as premature mortality. Available evidence indicates that 'stress-sensitive' chronic medical disorders, such as cardiovascular disease, obesity and Type 2 diabetes mellitus, are critical mediators and/or moderators of BD. Changes in physiologic systems implicated in allostasis have been proposed to impact brain structures and neurocognition, as well as medical comorbidity in this population. For example, abnormalities in insulin physiology, for example, insulin resistance, hyperinsulinemia and central insulinopenia, are implicated as effectors of allostatic load in BD. Insulin's critical role in CNS physiological (e.g., neurotrophism and synaptic plasticity) and pathophysiological (e.g., neurocognitive deficits, pro-apoptosis and amyloid deposition) processes is amply documented. This article introduces the concept that insulin is a mediator of allostatic load in the BD and possibly a therapeutic target.

Tumor necrosis factor-alpha mediates changes in tissue protein turnover in a rat cancer cachexia model.

Rats bearing the Yoshida AH-130 ascites hepatoma showed enhanced fractional rates of protein degradation in gastrocnemius muscle, heart, and liver, while fractional synthesis rates were similar to those in non-tumor bearing rats. This hypercatabolic pattern was associated with marked perturbations of the hormonal homeostasis and presence of tumor necrosis factor in the circulation. The daily administration of a goat anti-murine TNF IgG to tumor-bearing rats decreased protein degradation rates in skeletal muscle, heart, and liver as compared with tumor-bearing rats receiving a nonimmune goat IgG. The anti-TNF treatment was also effective in attenuating early perturbations in insulin and corticosterone homeostasis. Although these results suggest that tumor necrosis factor plays a significant role in mediating the changes in protein turnover and hormone levels elicited by tumor growth, the inability of such treatment to prevent a reduction in body weight implies that other mediators or tumor-related events were also involved.

Diabetes mellitus, periapical inflammation and endodontic treatment outcome

The possible connection between chronic oral inflammatory processes, such as apical periodontitis and periodontal disease (PD), and systemic health is one of the most interesting aspects faced by the medical and dental scientific community. Chronic apical periodontitis shares important characteristics with PD: 1) both are chronic infections of the oral cavity, 2) the Gram-negative anaerobic microbiota found in both diseases is comparable, and 3) in both infectious processes increased local levels of inflammatory mediators may have an impact on systemic levels. One of the systemic disorders linked to PD is diabetes mellitus (DM); is therefore plausible to assume that chronic apical periodontitis and endodontic treatment are also associated with DM. The status of knowledge regarding the relationship between DM and endodontics is reviewed. Upon review, we conclude that there are data in the literature that associate DM with a higher prevalence of periapical lesions, greater size of the osteolityc lesions, greater likelihood of asymptomatic infections and worse prognosis for root filled teeth. The results of some studies suggest that periapical disease may contribute to diabetic metabolic dyscontrol

Epithelial-to-mesenchymal transition mediates docetaxel resistance and high risk of relapse in prostate cancer

Molecular characterization of radical prostatectomy specimens after systemic therapy may identify a gene expression profile for resistance to therapy. This study assessed tumor cells from patients with prostate cancer participating in a phase II neoadjuvant docetaxel and androgen deprivation trial to identify mediators of resistance. Transcriptional level of 93 genes from a docetaxel-resistant prostate cancer cell lines microarray study was analyzed by TaqMan low-density arrays in tumors from patients with high-risk localized prostate cancer (36 surgically treated, 28 with neoadjuvant docetaxel þ androgen deprivation). Gene expression was compared between groups and correlated with clinical outcome. VIM, AR and RELA were validated by immunohistochemistry. CD44 and ZEB1 expression was tested by immunofluorescence in cells and tumor samples. Parental and docetaxel-resistant castration-resistant prostate cancer cell lines were tested for epithelial-to-mesenchymal transition (EMT) markers before and after docetaxel exposure. Reversion of EMT phenotype was investigated as a docetaxel resistance reversion strategy. Expression of 63 (67.7%) genes differed between groups (P < 0.05), including genes related to androgen receptor, NF-k B transcription factor, and EMT. Increased expression of EMT markers correlated with radiologic relapse. Docetaxel-resistant cells had increased EMT and stem-like cell markers expression. ZEB1 siRNA transfection reverted docetaxel resistance and reduced CD44 expression in DU-145R and PC-3R. Before docetaxel exposure, a selected CD44 þ subpopulation of PC-3 cells exhibited EMT phenotype and intrinsic docetaxel resistance; ZEB1/CD44 þ subpopulations were found in tumor cell lines and primary tumors; this correlated with aggressive clinical behavior. This study identifies genes potentially related to chemotherapy resistance and supports evi-dence of the EMT role in docetaxel resistance and adverse clinical behavior in early prostate cancer.

Mediació intercultural i institucions escolars: l'exemple del Quebec

L’increment de la multiplicitat cultural ha comportat que, en diversos països, s’elaborin diferents actuacions per aproximar posicions i millorar la comunicació entre progenitors i professionals de les institucions escolars, així com per facilitar l’adaptació del currículum a la diversitat cultural. Una de les opcions preses, amb cert èxit però no exempta d’errors,és la dels anomenats «mediadors interculturals». Una societat on aquesta figura, amb els noms d’agents de liaison i de milieu, s’ha desenvolupat és el Quebec. L’estudi dels seus encerts i limitacions, específicament a les institucions escolars, creiem que pot ser un referent a partir del qual cal reflexionar, atès el notable increment d’aquesta pràctica a Catalunya.

La figura del mediador intercultural a les institucions escolars catalanes: perfil real i ideal segons els propis actors

A Catalunya, en els darrers anys, han anat proliferant una sèrie de «professionals» que realitzen, amb més o menys adequació a la definició teòrica, la funció de mediadors inter-culturals. Primerament, es tractava de gitanos, però darrerament, fruit de l’increment de la presència d’infants d’origen immigrant a les escoles catalanes, s’ha vist créixer el nombre i el protagonisme dels estrangers pel reconeixement que se’ls ha donat des de l’Administració i des de les institucions. Els mediadors han intentat intervenir en qüestions com ara la participació dels pares d’origen minoritari, l’adaptació del currículum a la diversitat cultural, la negociació de conflictes culturals, la desescolarització, l’absentisme i l’abandonament escolar dels alumnes durant el període d’escolarització obligatòria, la traducció lingüística i la interpretació sociocultural, etc. Concretament, la nostra recerca ha consistit en vint-i-set entrevistes en profunditat a mediadors interculturals de tot Catalunya.

Actitud de los docentes respecto al mediador y la necesidad de formación en educación intercultural

En Cataluña, en los últimos años, han ido proliferando una serie de «profesionales» que realizan, con más o menos adecuación a la definición teórica, la función de mediadores interculturales. Primeramente se trataba de gitanos, pero recientemente, fruto del incremento de la presencia de niños de origen inmigrante en las escuelas catalanas, se ha visto crecer el número y el protagonismo de los extranjeros por el reconocimiento que se les ha otorgado desde la administración y las instituciones. Al ser una práctica nueva está aún llena de interrogantes, pero al notar que no se está produciendo paralelamente una reflexión y clarificación de sus fundamentos nos parecía un momento pertinente para examinar su desarrollo, por ello hemos realizado una encuesta al profesorado de enseñanza obligatoria (n=740) que nos han proporcionado su percepción.

Los docentes de Cataluña y el recurso del mediador intercultural

En Cataluña, en los últimos años, han ido proliferando una serie de "profesionales" que realizan, con más o menos adecuación a la definición teórica, la función de mediadores interculturales. Primeramente se trataba de gitanos, pero recientemente, fruto del incremento de la presencia de niños de origen inmigrante en las escuelas catalanas, se ha visto crecer el número y el protagonismo de los extranjeros por el reconocimiento que se les ha otorgado desde la administración y las instituciones. Al ser una práctica nueva está aún llena de interrogantes, pero al notar que no se está produciendo paralelamente una reflexión y clarificación de sus fundamentos nos parece que es un momento pertinente para examinar su desarrollo. Por ello hemos realizado una encuesta al profesorado de enseñanza obligatoria (n=740), que nos han proporcionado su percepción sobre la figura del mediador.