Reduction of cardiac TGFß-mediated profibrotic events by inhibition of Hsp90 with engineered protein.

Myocardial fibroblast activation coupled with extracellular matrix production is a pathological signature of myocardial fibrosis and is governed mainly by transforming growth factor TGFß-Smad2/3 signaling. Targeting the ubiquitous TGFß leads to cellular homeostasis deregulation with adverse consequences. We previously showed the anti-fibrotic effects upon downregulation of 90-kDa heat shock protein (Hsp90), a chaperone that associates to the TGFß signaling cascade. In the present study, we use a fluorescent-labeled Hsp90 protein inhibitor (CTPR390-488) with specific Hsp90 binding properties to reduce myocardial pro-fibrotic events in vitro and in vivo. The mechanism of action involves the disruption of TGFßRI-Hsp90 complex, resulting in a decrease in TGFß signaling and reduction in extracellular matrix collagen. In vivo, decreased myocardial collagen deposition was observed upon CTPR390-488 treatment in a pro-fibrotic mouse model. This is the first study demonstrating the ability of an engineered Hsp90 protein inhibitor to block collagen expression, reduce the motility of myocardial TGFß-activated fibroblasts and ameliorate angiotensin-II induced cardiac myocardial fibrosis in vivo.

Correlative 3D cryo X-ray imaging reveals intracellular location and effect of designed antifibrotic protein-nanomaterial hybrids.

Revealing the intracellular location of novel therapeutic agents is paramount for the understanding of their effect at the cell ultrastructure level. Here, we apply a novel correlative cryo 3D imaging approach to determine the intracellular fate of a designed protein-nanomaterial hybrid with antifibrotic properties that shows great promise in mitigating myocardial fibrosis. Cryo 3D structured illumination microscopy (cryo-3D-SIM) pinpoints the location and cryo soft X-ray tomography (cryo-SXT) reveals the ultrastructural environment and subcellular localization of this nanomaterial with spatial correlation accuracy down to 70 nm in whole cells. This novel high resolution 3D cryo correlative approach unambiguously locates the nanomaterial after overnight treatment within multivesicular bodies which have been associated with endosomal trafficking events by confocal microscopy. Moreover, this approach allows assessing the cellular response towards the treatment by evaluating the morphological changes induced. This is especially relevant for the future usage of nanoformulations in clinical practices. This correlative super-resolution and X-ray imaging strategy joins high specificity, by the use of fluorescence, with high spatial resolution at 30 nm (half pitch) provided by cryo-SXT in whole cells, without the need of staining or fixation, and can be of particular benefit to locate specific molecules in the native cellular environment in bio-nanomedicine.

Topical interferon alpha-2B topic as the first therapeutic option in a clinical case of conjunctival intraepithelial neoplasia. / Interferón alfa-2B tópico como primera opción terapéutica en un caso clínico de neoplasia intraepitelial córneo-conjuntival.

INTRODUCTION: Conjunctival intraepithelial neoplasia is a pre-malignant lesion of the ocular surface. It can be treated with topical interferon alpha-2b (INF α-2b) as first choice. CLINICAL CASE: A 71-year-old man referred for corneal-conjunctival, gelatinous lesion in the left eye (LE) with an area of almost 270°. The clinical diagnosis was compatible with a corneal-conjunctival intraepithelial neoplasia. Topical treatment was started with INF α-2b at a dose of one million international units (IU)/ml, 4 times/day for 4 months, with remission being achieved. CONCLUSION: The isolated use of topical INF α-2b is an effective treatment as a first option in the case of corneal-conjunctival intraepithelial neoplasia, positioning itself as a form of effective and safe treatment compared to other therapeutic options. Surgical excision and use of other chemotherapy agents could lead to severe limbic deficits and other side effects.

Validation of a hybrid Doppler ultrasound vessel-based registration algorithm for neurosurgery.

PURPOSE: We describe and validate a novel hybrid nonlinear vessel registration algorithm for intra-operative updating of preoperative magnetic resonance (MR) images using Doppler ultrasound (US) images acquired on the dura for the correction of brain-shift and registration inaccuracies. We also introduce an US vessel appearance simulator that generates vessel images similar in appearance to that acquired with US from MR angiography data. METHODS: Our registration uses the minimum amount of preprocessing to extract vessels from the raw volumetric images. This prevents the removal of important registration information and minimizes the introduction of artifacts that may affect robustness, while reducing the amount of extraneous information in the image to be processed, thus improving the convergence speed of the algorithm. We then completed 3 rounds of validation for our vessel registration method for robustness and accuracy using (i) a large number of synthetic trials generated with our US vessel simulator, (ii) US images acquired from a real physical phantom made from polyvinyl alcohol cryogel, and (iii) real clinical data gathered intra-operatively from 3 patients. RESULTS: Resulting target registration errors (TRE) of less than 2.5 mm are achieved in more than 90 % of the synthetic trials when the initial TREs are less than 20 mm. TREs of less than 2 mm were achieved when the technique was applied to the physical phantom, and TREs of less than 3 mm were achieved on clinical data. CONCLUSIONS: These test trials show that the proposed algorithm is not only accurate but also highly robust to noise and missing vessel segments when working with US images acquired in a wide range of real-world conditions.

Interferón alfa-2B tópico como primera opción terapéutica en un caso clínico de neoplasia intraepitelial córneo-conjuntival / Topical interferon alpha-2B topic as the first therapeutic option in a clinical case of conjunctival intraepithelial neoplasia

Introducción: La neoplasia intraepitelial córneo-conjuntival es una lesión premaligna de la superficie ocular. Puede ser tratada con interferón alfa-2b (INF α-2b) tópico como tratamiento de primera elección. Caso clínico: Varón de 71 años de edad remitido por lesión córneo-conjuntival en ojo izquierdo (OI) de aspecto gelatinoso con una extensión de casi 270° (de las IX a las V h), cuyo diagnóstico clínico era compatible con una neoplasia intraepitelial córneo-conjuntival. Se inició tratamiento tópico con INF α-2b, en dosis de un millón de unidades internacionales (UI)/mililitro (ml), con una posología de 4 veces/día durante 4 meses, con lo que se consiguió su remisión. Conclusión: El uso aislado de INF α-2b tópico es un tratamiento efectivo como primera opción en el caso de neoplasia intraepitelial córneo-conjuntival, que se sitúa como una forma de tratamiento eficaz y segura frente a otras opciones terapéuticas como la escisión quirúrgica y frente al empleo de otros agentes quimioterápicos que pueden condicionar insuficiencias límbicas severas, entre otros efectos secundarios (AU)

Introduction: Conjunctival intraepithelial neoplasia is a pre-malignant lesion of the ocular surface. It can be treated with topical interferon alpha-2b (INF α-2b) as first choice. Clinical case: A 71-year-old man referred for corneal-conjunctival, gelatinous lesion in the left eye (LE) with an area of almost 270°. The clinical diagnosis was compatible with a corneal-conjunctival intraepithelial neoplasia. Topical treatment was started with INF α-2b at a dose of one million international units (IU)/ml, 4 times/day for 4 months, with remission being achieved. Conclusion: The isolated use of topical INF α-2b is an effective treatment as a first option in the case of corneal-conjunctival intraepithelial neoplasia, positioning itself as a form of effective and safe treatment compared to other therapeutic options. Surgical excision and use of other chemotherapy agents could lead to severe limbic deficits and other side effects (AU)