Hair follicle stem cell differentiation is inhibited through cross-talk between Wnt/ß-catenin and androgen signalling in dermal papilla cells from patients with androgenetic alopecia.

BACKGROUND: Hair follicle (HF) regeneration begins when signals from the mesenchyme-derived dermal papilla cells (DPC) reach multipotent epidermal stem cells in the bulge region. Wnt/ß-catenin signalling is known to affect mammalian hair growth positively. In androgenetic alopecia (AGA), androgens cause HF miniaturization through a mechanism that remains unclear. Circulating androgens act on DPC and alter paracrine factors that influence hair epithelial cells. OBJECTIVES: To elucidate the role of androgens in dermal papilla-induced differentiation of HF stem cells. METHODS: HF stem cell differentiation was evaluated in a coculture model with DPC or culturing with media conditioned by DPC after activation of androgen and Wnt/ß-catenin signalling pathways. To study the molecular cross-talk between the androgen and Wnt signalling pathway in DPC, we analysed the expression and activation of downstream Wnt signalling molecules in the presence of androgens. RESULTS: In a coculture model with human DPC from patients with AGA and HF stem cells, we observed that androgens abrogate hair differentiation evaluated by hair-specific keratin 6 expression. Wnt signalling activation restored the ability of androgen-treated DPC to induce differentiation. Androgen treatment revealed a significant decrease in the cytoplasmic/total ß-catenin protein ratio and upregulation of the activity of glycogen synthase kinase-3ß in DPC, indicative of canonical Wnt pathway inhibition. CONCLUSIONS: These results suggest that androgens deregulate DPC-secreted factors involved in normal HF stem cell differentiation via the inhibition of the canonical Wnt signalling pathway.

Metastatic cancer stem cells: new molecular targets for cancer therapy.

The cancer stem cell (CSC) hypothesis, predicts that a small subpopulation of cancer cells that possess "stem-like" characteristics, are responsible for initiating and maintaining cancer growth. According to the CSC model the many cell populations found in a tumour might represent diverse stages of differentiation. From the cellular point of view metastasis is considered a highly inefficient process and only a subset of tumour cells is capable of successfully traversing the entire metastatic cascade and eventually re-initiates tumour growth at distant sites. Some similar features of both normal and malignant stem cells suggest that CSCs are not only responsible for tumorigenesis, but also for metastases. The CSC theory proposes that the ability of a tumour to metastasize is an inherent property of a subset of CSCs. The similar biological characteristics shared by normal stem cells (NSCs) and CSCs mainly implicate self-renewal and differentiation potential, survival ability, niche-specific microenvironment requirements and specific homing to metastatic sites and may have important implications in terms of new approaches to cancer therapy in the metastatic setting. There are several agents targeting many of these CSC features that have shown to be effective both in vitro and in vivo. Although clinical trials results are still preliminary and continue under investigation, these new therapies are very promising. The identification of new therapeutic targets and drugs based on CSC model constitutes a great challenge.

EDUCA study: Psychometric properties of the Spanish version of the Zarit Caregiver Burden Scale.

OBJECTIVES: The Zarit Caregiver Burden Scale, translated and validated into Spanish, is sensitive to the application of a Psychoeducational Intervention Program (PIP) for the prevention and reduction of burden in caregivers of Alzheimer's disease (AD) patients (EDUCA study). The data obtained in EDUCA was used to reanalyse its psychometric properties and the cut-off points of the Zarit scale. METHODS: The scale was administered to 115 caregivers of patients with AD who were randomised to receive a PIP or standard care for four months. Internal reliability and a factorial analysis of principal components were assessed, and the impact of PIP on each of the subscales was evaluated. A cut-off point was defined for the Zarit scale to identify the caregivers most sensitive to receiving a PIP. RESULTS: A good internal reliability (Cronbach alpha coefficient of 0.92) was obtained, with three principal components (burden, competency and dependence) explaining 54.75% of the variance. The application of PIP showed statistically significant differences versus standard care for the dependence subscale (p = 0.0082) (p = 0.062 for the burden scale). The Zarit scale cut-off points which combine better sensitivity and specificity were 56/57 and 59/60, for the 5/6 and 6/7 cut-off points of the General Health Questionnaire (GHQ-28) scale, respectively. CONCLUSIONS: This study confirms the good psychometric properties of the Zarit scale found in previous studies. The dependence component appeared to be most influenced by the application of a PIP in the clinical trial. Caregivers with a Zarit scale score of 60 or more benefit most from the PIP.

[Observational study to evaluate the impact of an educational/informative intervention in the emotional status (anxiety) of patients with atopic dermatitis (CUIDA-DEL)]. / Estudio observacional para evaluar la repercusión de una intervención educativa-informativa en el estado emocional (ansiedad) de los pacientes con dermatitis atópica (CUIDA-DEL).

INTRODUCTION: One of the first therapeutic measures in atopic dermatitis should be the educational and informative approach about prophylactic aspects and evolution of the disease. This type of proceedings has been shown to be beneficial for the anxious type of emotional status in patients with atopic dermatitis. We evaluated the impact of an educational/informative intervention in the emotional status (anxiety) of patients with atopic dermatitis in Spain. MATERIAL AND METHODS: Investigators were randomized into two study groups: the control group (CG) that followed current clinical practice and the intervention group (IG) that handed patients, in each visit, a booklet of information about different prophylactic aspects and care of atopic dermatitis. The duration of the study was 6 months with quarterly visits. All included patients had a diagnosis of atopic dermatitis. Anxiety was evaluated with the STAI anxiety questionnaire and clinical data regarding dermatological aspects (IGA, pruritus scale, location of the lesions) were also recorded. RESULTS: A total of 1,247 patients were recruited thanks to the collaboration of 158 investigators. Patients were distributed as follows: 683 (54.7 %) in the CG and 564 (45.2 %) in the IG. Both group were homogeneous with respect to basal characteristics, and were constituted by 54 % of women with a mean age of 19 years. Eighty-six percent of atopic dermatitis lesions were preferentially located in extremities. Patients of both study groups showed improvement in their emotional status (trait and state anxiety) throughout the study with significant decreases in the STAI scores compared to basal ones. Regarding improvement in the questionnaire scores, no significant differences were observed between groups except in children aged 9 to 15 years, in the pediatric version of the STAI trait where the percentage of score decrease at 6 months adjusted to the basal score was 5.5 (19.0) for the CG and 10.6 (18.9) for the IG, (p < 0.05). A higher percentage of patients finished the study in the IG compared to the CG (83.1 % vs. 76.1 % p < 0.005). DISCUSSION AND CONCLUSIONS: Although patients in the IG showed greater compliance with the follow-up of the study, the informative intervention about prophylactic aspects of atopic dermatitis designed in this study does not appear to have had an impact in improving the emotional status of adult patients.

Estudio observacional para evaluar la repercusión de una intervención educativa-informativa en el estado emocional (ansiedad) de los pacientes con dermatitis atópica (CUIDA-DEL) / Observational study to evaluate the impact of an educational/informative intervention in the emotional status (anxiety) of patients with atopic dermatitis (CUIDA-DEL)

Introducción. Una de las primeras actuaciones terapéuticas en dermatitis atópica (DA) debe ser el abordaje educacional e informativo sobre los aspectos preventivos y de evolución de la enfermedad. Actuaciones de este tipo han demostrado ser beneficiosas en el estado emocional de tipo ansioso de los pacientes con DA. Se evaluó la repercusión de una intervención educativa-informativa en el estado emocional (ansiedad) de pacientes afectados de DA en España. Material y métodos. Se aleatorizaron a los investigadores en dos grupos de estudio: control (GC), que siguió práctica clínica habitual y grupo intervención (GI) que entregó, en cada visita, un boletín informativo sobre diferentes aspectos preventivos y de cuidados de la DA a los pacientes. La duración del estudio fue de 6 meses con visitas trimestrales. Todos los pacientes incluidos tenían diagnóstico de DA. La ansiedad se valoró con los cuestionarios de ansiedad State Trait Anxiety Inventory (STAI) y se recogieron datos clínicos de evolución dermatológicos: IGA, escala de prurito y localización de la DA. Resultados. Se reclutaron 1.247 pacientes gracias a la colaboración de 158 investigadores. Los grupos de pacientes se distribuyeron: 683 (54,7 %) en el GC y 564 (45,2 %) en el GI. Ambos grupos fueron homogéneos basalmente, constituidos por un 54 % de mujeres, con una edad media de 19 años. El 86 % de la DA estuvo localizada preferentemente en extremidades. Los pacientes de ambos grupos del estudio mejoraron en su estado emocional (ansiedad rasgo y estado) a lo largo del mismo con descensos significativos en las puntuaciones del STAI respecto a la basal. No se apreciaron diferencias significativas entre los grupos en la mejoría del cuestionario, excepto en niños de 9 a 15 años, en la versión infantil del STAI rasgo, donde el porcentaje de descenso, de la puntuación a los 6 meses corregida por la puntuación basal, fue para el GC de 5,5 (19,0) y para el GI de 10,6 (18,9) (p < 0,05). En el GI un mayor porcentaje de pacientes finalizaron el estudio comparado con el GC (83,1 % frente a 76,1 % p < 0,005). Discusión y conclusiones. A pesar de que en el GI se consiguió una adherencia mayor de los pacientes en el seguimiento del estudio, la intervención informativa sobre hábitos de prevención de DA diseñada en este estudio no parece haber tenido una repercusión significativa en la mejoría del estado emocional de los pacientes adultos

Introduction. One of the first therapeutic measures in atopic dermatitis should be the educational and informative approach about prophylactic aspects and evolution of the disease. This type of proceedings has been shown to be beneficial for the anxious type of emotional status in patients with atopic dermatitis. We evaluated the impact of an educational/informative intervention in the emotional status (anxiety) of patients with atopic dermatitis in Spain. Material and methods. Investigators were randomized into two study groups: the control group (CG) that followed current clinical practice and the intervention group (IG) that handed patients, in each visit, a booklet of information about different prophylactic aspects and care of atopic dermatitis. The duration of the study was 6 months with quarterly visits. All included patients had a diagnosis of atopic dermatitis. Anxiety was evaluated with the STAI anxiety questionnaire and clinical data regarding dermatological aspects (IGA, pruritus scale, location of the lesions) were also recorded. Results. A total of 1,247 patients were recruited thanks to the collaboration of 158 investigators. Patients were distributed as follows: 683 (54.7 %) in the CG and 564 (45.2 %) in the IG. Both group were homogeneous with respect to basal characteristics, and were constituted by 54 % of women with a mean age of 19 years. Eighty-six percent of atopic dermatitis lesions were preferentially located in extremities. Patients of both study groups showed improvement in their emotional status (trait and state anxiety) throughout the study with significant decreases in the STAI scores compared to basal ones. Regarding improvement in the questionnaire scores, no significant differences were observed between groups except in children aged 9 to 15 years, in the pediatric version of the STAI trait where the percentage of score decrease at 6 months adjusted to the basal score was 5.5 (19.0) for the CG and 10.6 (18.9) for the IG, (p < 0.05). A higher percentage of patients finished the study in the IG compared to the CG (83.1 % vs. 76.1 % p < 0.005). Discussion and conclusions. Although patients in the IG showed greater compliance with the follow-up of the study, the informative intervention about prophylactic aspects of atopic dermatitis designed in this study does not appear to have had an impact in improving the emotional status of adult patients

[Effectiveness of an intervention to provide information to patients with hypertension as short text messages and reminders sent to their mobile phone (HTA-Alert)]. / Eficacia de una intervención informativa a hipertensos mediante mensajes de alerta en el teléfono móvil (HTA-ALERT).

OBJECTIVE: To analyze the effect of an intervention to provide information with mobile phone text messages to patients with hypertension on compliance with therapy for hypertension. DESIGN: Comparative, controlled, multicenter, randomized cluster study. SETTING: 26 primary care health centers in Spain. PARTICIPANTS: 26 researchers were randomized to a control group or an intervention group (52 patients each, for a total of 104 patients). All patients were receiving monotherapy for uncontrolled hypertension. INTERVENTION: Patients in the control group received their physician's usual interventions. Patients in the intervention group received messages and reminders sent to their mobile phones 2 days per week during 4 months. MAIN OUTCOME MEASURES: Tablets were counted and blood pressure was measured at the start of the study and 1, 3, and 6 months later. The percentage of compliers, mean percentage of compliance and degree of control of hypertension were compared. The reduction in absolute and relative risk was calculated, as was the number of individuals needed to treat to avoid noncompliance. RESULTS: The results were evaluated for a total of 67 individuals (34 in the intervention group and 33 in the control group). The rate of compliance was 85.1% (CI, 74.9%-95.3%) overall, 85.7% (CI, 70.5%-100.9%) in the control group and 84.4% in the intervention group (CI, 70.7%-95.3%) (P=NS). Mean percentage compliance was 90.2%+/-16.3% overall, 88.1%+/-20.8% in the control group and 91.9%+/-11.6% in the intervention group (P=NS). The percentage of patients whose hypertension was controlled at the end of the study was 51.5% (CI, 34.4%-68.6%) in the control group and 64.7% (CI, 48.6%-80.8%) in the intervention group (P=NS). CONCLUSIONS: The telephone messaging intervention with alerts and reminders sent to mobile phones did not improve compliance with therapy in patients with hypertension.

Antiandrogen oligonucleotides: active principles in hair- and skin-derived culture cells.

Most drugs used for treatment of androgen-related dermatological disorders are not completely satisfactory in terms of clinical efficacy and potential secondary effects. There is, therefore, a need for a new generation of specific antiandrogens. This paper focuses on an oligonucleotide antisense pharmacological strategy. Acceptor sites were first disclosed by mapping the human Androgen Receptor (AR) mRNA conformation using an mRNA walking approach, oligonucleotide binding, and S1 protection assays. Antisense-sensitive regions were localized by RNAse H degradation and AR in vitro translation inhibition. Oligonucleotides were then designed and assessed, in primary cultures of human hair dermal papillae and skin derived fibroblasts, for their capability to down-regulate AR expression. Some of them were able to inhibit more than 60 to 80% of the AR expression. These could be a new class of antiandrogen oligonucleotides pharmacologically active in hair and skin derived cells, suitable for the treatment of dermatological disorders.

Activation of ErbB-2 via a hierarchical interaction between ErbB-2 and type I insulin-like growth factor receptor in mammary tumor cells.

The present study focused on interactions between signaling pathways activated by progestins and by type I and II receptor tyrosine kinases (RTKs) in mammary tumors. An experimental model in which the synthetic progestin medroxyprogesterone acetate (MPA) induced mammary adenocarcinomas in Balb/c mice was used. MPA-stimulated proliferation, both in vivo and in vitro, of progestin-dependent tumors induced up-regulation of ErbB-2 protein levels and tyrosine phosphorylation of this receptor. Combinations of antisense oligodeoxynucleotides (ASODNs) directed to ErbB-2 mRNA with ASODNs directed to the insulin-like growth factor-I receptor (IGF-IR) were used to study the effect of the simultaneous block of these receptors on the MPA-induced proliferation of epithelial cells from the progestin-dependent C4HD line. Neither synergistic nor additive effects on the inhibition of MPA-induced proliferation of C4HD cells were observed as a result of the combination of these ASODNs. Suppression of IGF-IR expression by ASODNs resulted in complete abrogation of MPA-induced phosphorylation of ErbB-2 in C4HD cells, whereas blockage of ErbB-2 did not affect IGF-IR phosphorylation. These results show the existence of a hierarchical interaction between IGF-IR and ErbB-2, by means of which IGF-IR directs ErbB-2 phosphorylation. We demonstrated, for the first time, that this hierarchical interaction involves physical association of both receptors, resulting in the formation of a heteromeric complex. Furthermore, confocal laser microscopy experiments demonstrated that MPA was able to induce co-localization of ErbB-2 and IGF-IR. This hetero-oligomer was also found in MCF-7 human breast cancer cells in which association of IGF-IR and ErbB-2 was induced by heregulin and IGF-I. Oncogene (2001) 20, 34 - 47.

Interactions between progestins and heregulin (HRG) signaling pathways: HRG acts as mediator of progestins proliferative effects in mouse mammary adenocarcinomas.

The present study addressed links between progestin and heregulin (HRG) signaling pathways in mammary tumors. An experimental model of hormonal carcinogenesis, in which the synthetic progestin medroxyprogesterone acetate (MPA) induced mammary adenocarcinomas in female Balb/c mice, was used. MPA induced an in vivo up-regulation of HRG mRNA expression in progestin-dependent (HD) tumor lines. Mammary tumor progression to a progestin-independent (HI) phenotype was accompanied by a high constitutive expression of HRG. The HRG message arose from the tumor epithelial cells. Primary cultures of malignant epithelial cells from a HD tumor line were used to investigate HRG involvement on cell proliferation. HRG induced a potent proliferative effect on these cells and potentiated MPA mitogenic effects. Blocking endogenous HRG synthesis by antisense oligodeoxynucleotides (ASODNs) to HRG mRNA inhibited MPA-induced cell growth, indicating that HRG acts as a mediator of MPA-induced growth. High levels of ErbB-2 and ErbB-3 expression and low ErbB-4 levels were found in HD cells. Treatment of these cells with either MPA or HRG resulted in tyrosine phosphorylation of both ErbB-2 and ErbB-3. Furthermore, both HRG and MPA proliferative effects were abolished when cells were treated with ASODNs to ErbB-2 mRNA, providing evidence for a critical role of ErbB-2 in HRG-induced growth. Finally, blocking type I insulin-like growth factor receptor (IGF-IR) expression with ASODN resulted in the complete inhibition of HRG proliferative effect, demonstrating that a functional IGF-IR is required for HRG mitogenic activity. These results provide the first evidence of interactions between progestins and HRB/ErbB signal transduction pathways in mammary cancer and the first demonstration that IGF-IR is required for HRG proliferative effects.

Involvement of insulin-like growth factors-I and -II and their receptors in medroxyprogesterone acetate-induced growth of mouse mammary adenocarcinomas.

The role of the insulin-like growth factors (IGFs) system was investigated in hormone-dependent (HD) and -independent (HI) in vivo lines of the medroxyprogesterone acetate (MPA)-induced mammary tumor model in Balb/c mice. IGF-II protein and message showed a three- to four-fold increase in HD lines growing in MPA-treated mice, as compared with HD tumors growing in untreated mice. Progression to a hormone-independent phenotype in all these lines was accompanied by a high constitutive expression of IGF-II. Similar IGF-I mRNA levels were detected in HD and HI lines. Both IGF-I and -II messages arose from the malignant epithelial cells, as shown by in situ hybridization studies. A significant decrease in Man-6P/type II IGF-R content was detected in HD tumors growing in MPA-treated mice as compared with HD lines growing in untreated mice. On the other hand, in HI tumors, notwithstanding high IGF-II synthesis, the levels of Man-6P/type II IGF-R remain high. Competitive inhibition and affinity labeling studies showed an almost exclusive binding of IGF-II to Man-6P/type II IGF-R on tumor membranes. The involvement of IGFs in the growth of epithelial primary cultures of the C4-HD line was evaluated. Exogenous IGF-I potentiated MPA stimulatory effect at concentrations of 50-100 ng/ml. Treatment of C4-HD cells with antisense oligodeoxynucleotides (ASODNs) to type I IGF-R and to IGF-II RNA resulted in a dose-dependent inhibition of MPA-mediated cell proliferation. The inhibition caused by IGF-II ASODNs could not be overcome by the addition of IGF-II up to 150 ng/ml. ASODNs to type I IGF-R at 40 microg/ml reduced by 75% the number of type I IGF-R; ASODNs to IGF-II at 1 microM decreased by 83% the levels of IGF-II protein. Our results provide support for the involvement of IGF-I and -II in MPA-induced mammary tumor growth by autocrine pathways.

[Growth hormones and oncogenes in mammary adenocarcinomas induced by medroxyprogesterone acetate in BALB/c mice]. / Factores de crecimiento y oncogenes en adenocarcinomas mamarios inducidos por acetato de medroxyprogesterona en ratones BALB/c.

We have studied the involvement of growth factors (GF), their receptors (GF-R) and oncogenes in modulating tumor growth in the medroxyprogesterone acetate (MPA)-induced mammary tumor model in BALB/c mice. We demonstrated the presence of both ligands of the insulin-like growth factor family (IGF-I, IGF-II) and the two types of receptors (IGF-RI, IGF-RII). MPA upregulated IGF-II mRNA and protein levels in hormone-dependent lines (MPA-D). The progression to a hormone-independent phenotype was accompanied by a high constitutive expression of IGF-II and by a significant decrease in IGF-IIR number. An antisense strategy used to evaluate the role of IGF in the MPA-induced growth of epithelial MPA-D cells showed that IGF mediate progestin-induced mammary tumor growth by autocrine/intracrine pathways. We also studied the role of heregulins (HRG), the recently identified ligands for the c-erbB3 and c-erbB4 oncogenes. HRG mRNA expression was restricted to tumors of ductal origin. MPA induced an in vivo up-regulation of HRG expression. Finally, we also found that MPA may be exerting its proliferative effect on MPA-D lines by inhibiting the expression of transforming growth factor beta 1, (TGF-beta 1) and the lack of expression of TGF-beta 1 in hormone-independent tumors may be related to the acquisition of autonomous growth.