Fluorescence-Coupled Techniques for Determining Rose Bengal in Dermatological Formulations and Their Application to Ex Vivo Skin Deposition Studies

Rose Bengal (RB) is a fluorescent dye with several potential biomedical applications, particularly in dermatology. Due to RB’s poor physicochemical properties, several advanced delivery systems have been developed as a potential tool to promote its permeation across the skin. Nevertheless, no validated quantitative method to analyse RB within the skin is described in the literature. Considering RB exhibits a conjugated ring system, the current investigation proposes fluorescence-based techniques beneficial for qualitatively and quantitatively determining RB delivered to the skin. Notably, the development and validation of a fluorescence-coupled HPLC method to quantify RB within the skin matrix are herein described for the first time. The method was validated based on the ICH, FDA and EMA guidelines, and the validated parameters included specificity, linearity, LOD, LLOQ, accuracy and precision, and carry-over and dilution integrity. Finally, the method was applied to evaluate RB’s ex vivo permeation and deposition profiles when loaded into dermatological formulations. Concerning qualitative determination, multiphoton microscopy was used to track the RB distribution within the skin strata, and fluorescence emission spectra were investigated to evaluate RB’s behaviour when interacting with different environments. The analytical method proved specific, precise, accurate and sensitive to analyse RB in the skin. In addition, qualitative side-analytical techniques were revealed to play an essential role in evaluating the performance of RB’s dermatological formulation

Inhibitory effects of megakaryocytic cells in prostate cancer skeletal metastasis

Prostate cancer cells commonly spread through the circulation, but few successfully generate metastatic foci in bone. Osteoclastic cellular activity has been proposed as an initiating event for skeletal metastasis. Megakaryocytes (MKs) inhibit osteoclastogenesis, which could have an impact on tumor establishment in bone. Given the location of mature MKs at vascular sinusoids, they may be the first cells to physically encounter cancer cells as they enter the bone marrow. Identification of the interaction between MKs and prostate cancer cells was the focus of this study. K562 (human MK precursors) and primary MKs derived from mouse bone marrow hematopoietic precursor cells potently suppressed prostate carcinoma PC-3 cells in coculture. The inhibitory effects were specific to prostate carcinoma cells and were enhanced by direct cell-cell contact. Flow cytometry for propidium iodide (PI) and annexin V supported a proapoptotic role for K562 cells in limiting PC-3 cells. Gene expression analysis revealed reduced mRNA levels for cyclin D1, whereas mRNA levels of apoptosis-associated specklike protein containing a CARD (ASC) and death-associated protein kinase 1 (DAPK1) were increased in PC-3 cells after coculture with K562 cells. Recombinant thrombopoietin (TPO) was used to expand MKs in the marrow and resulted in decreased skeletal lesion development after intracardiac tumor inoculation. These novel findings suggest a potent inhibitory role of MKs in prostate carcinoma cell growth in vitro and in vivo. This new finding, of an interaction of metastatic tumors and hematopoietic cells during tumor colonization in bone, ultimately will lead to improved therapeutic interventions for prostate cancer patients. © 2011 American Society for Bone and Mineral Research.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/78486/1/204_ftp.pd

Network Slices for Vertical Industries

Network Slicing allows to simultaneously support the specific needs of vertical industries with a diverse range of networking and computing requirements. Network Functions Virtualization (NFV) has been defined to deploy multiple network services on a common infrastructure. We extend the NFV concept to vertical services, i.e. services implemented on top of network services and providing the applications of the verticals. We present a component of the 5G-Transformer system, named vertical slicer, which acts as the interface to verticals. The vertical slicer has two main functionalities: allowing verticals to define vertical services based on a set of service blueprints and arbitrating among several vertical services in case of resource shortage.This work has been partially funded by the EU H2020 5G-Transformer Project (grant no. 761536

Automated service provisioning and hierarchical SLA management in 5G systems

Empowered by network softwarization, 5G systems have become the key enabler to foster the digital transformation of the vertical industries by expanding the scope of traditional mobile networks and enriching the network service offerings. To make this a reality, we propose an automation solution for vertical services provisioning and hierarchical Service Level Agreement (SLA) management. Service scaling is one of the most essential operations to adapt the service deployments and resource allocations to ensure SLA fulfilment. Three different scaling levels are addressed in this work: application-, service- and resource-level. We have implemented our solution in a proof-of-concept of a virtualized mobile network platform, spanning over three geographically-distributed sites. To evaluate our solution, we leverage field tests, focusing on automotive vertical services comprising a mission-critical application (collision-avoidance) and an entertainment one (video streaming). The results demonstrate the excellent performance of our solution, and its ability to automatically deploy vertical services and ensure their SLAs through different levels of service scaling.This work has been partially supported by European Commission H2020 5GPPP through the 5G-TRANSFORMER and 5GROWTH projects (Grants No. 761536 and 856709)

An atlas of cortical circular RNA expression in Alzheimer disease brains demonstrates clinical and pathological associations.

Parietal cortex RNA-sequencing (RNA-seq) data were generated from individuals with and without Alzheimer disease (AD; ncontrol = 13; nAD = 83) from the Knight Alzheimer Disease Research Center (Knight ADRC). Using this and an independent (Mount Sinai Brain Bank (MSBB)) AD RNA-seq dataset, cortical circular RNA (circRNA) expression was quantified in the context of AD. Significant associations were identified between circRNA expression and AD diagnosis, clinical dementia severity and neuropathological severity. It was demonstrated that most circRNA-AD associations are independent of changes in cognate linear messenger RNA expression or estimated brain cell-type proportions. Evidence was provided for circRNA expression changes occurring early in presymptomatic AD and in autosomal dominant AD. It was also observed that AD-associated circRNAs co-expressed with known AD genes. Finally, potential microRNA-binding sites were identified in AD-associated circRNAs for miRNAs predicted to target AD genes. Together, these results highlight the importance of analyzing non-linear RNAs and support future studies exploring the potential roles of circRNAs in AD pathogenesis

Novel lipid nanovesicle-loaded dissolving microarray patches for fenretinide in breast cancer chemoprevention

The retinoid fenretinide (FENR) is a promising compound for preventing breast cancer recurrence but faceschallenges due to poor solubility and low bioavailability. This study explores the development of dissolvingmicroneedles (MNs) containing FENR-loaded ethosomes for minimally invasive breast cancer chemoprevention,aiming to enhance local drug distribution. Ethosomes were formulated using ethanol, propylene glycol, soyalecithin, water, and polysorbate 80 micelles. MNs were created from poly(vinyl alcohol) and poly(vinylpyrrolidone) hydrogels by adding polymer powder directly into ethosomes suspensions, reducing manufacturingtime and cost. Two methods were used to load ethosomes into high-density moulds: 1) only in the needle area,and 2) in both the needle area and baseplate. Dynamic light scattering confirmed nanostructures in the hydrogelsand MNs. Micelle-based ethosomes dissolved MNs in 15 min, compared to 30 min for other MNs. Skin depositionstudies showed greater drug deposition (up to 10 μg/patch) and enhanced skin permeation of FENR (up to 40 μg)with Method 2. In-vivo studies in rats demonstrated that oral administration resulted in plasma FENR levelsbelow 10 ng/g in the first three hours, whereas MN administration delayed delivery, reaching a maximumplasma concentration of 52 ng/g at 48 h. Skin deposition of FENR from MNs decreased from 3 μg/g on day 1 to<0.3 μg/g by the last day. This study indicates that MNs are a potential minimally invasive dosage form fordelivering FENR, offering a new approach for breast cancer chemoprevention

Infections with Avian Pathogenic and Fecal Escherichia coli Strains Display Similar Lung Histopathology and Macrophage Apoptosis

The purpose of this study was to compare histopathological changes in the lungs of chickens infected with avian pathogenic (APEC) and avian fecal (Afecal) Escherichia coli strains, and to analyze how the interaction of the bacteria with avian macrophages relates to the outcome of the infection. Chickens were infected intratracheally with three APEC strains, MT78, IMT5155, and UEL17, and one non-pathogenic Afecal strain, IMT5104. The pathogenicity of the strains was assessed by isolating bacteria from lungs, kidneys, and spleens at 24 h post-infection (p.i.). Lungs were examined for histopathological changes at 12, 18, and 24 h p.i. Serial lung sections were stained with hematoxylin and eosin (HE), terminal deoxynucleotidyl dUTP nick end labeling (TUNEL) for detection of apoptotic cells, and an anti-O2 antibody for detection of MT78 and IMT5155. UEL17 and IMT5104 did not cause systemic infections and the extents of lung colonization were two orders of magnitude lower than for the septicemic strains MT78 and IMT5155, yet all four strains caused the same extent of inflammation in the lungs. The inflammation was localized; there were some congested areas next to unaffected areas. Only the inflamed regions became labeled with anti-O2 antibody. TUNEL labeling revealed the presence of apoptotic cells at 12 h p.i in the inflamed regions only, and before any necrotic foci could be seen. The TUNEL-positive cells were very likely dying heterophils, as evidenced by the purulent inflammation. Some of the dying cells observed in avian lungs in situ may also be macrophages, since all four avian E. coli induced caspase 3/7 activation in monolayers of HD11 avian macrophages. In summary, both pathogenic and non-pathogenic fecal strains of avian E. coli produce focal infections in the avian lung, and these are accompanied by inflammation and cell death in the infected areas