A cell level automated approach for quantifying antibody staining in immunohistochemistry images. A structural approach for quantifying antibody staining in colonic cancer spheroid images by integrating image processing and machine learning towards the implementation of computer aided scoring of cancer markers.

Immunohistological (IHC) stained images occupy a fundamental role in the pathologist¿s diagnosis and monitoring of cancer development. The manual process of monitoring such images is a subjective, time consuming process that typically relies on the visual ability and experience level of the pathologist. A novel and comprehensive system for the automated quantification of antibody inside stained cell nuclei in immunohistochemistry images is proposed and demonstrated in this research. The system is based on a cellular level approach, where each nucleus is individually analyzed to observe the effects of protein antibodies inside the nuclei. The system provides three main quantitative descriptions of stained nuclei. The first quantitative measurement automatically generates the total number of cell nuclei in an image. The second measure classifies the positive and negative stained nuclei based on the nuclei colour, morphological and textural features. Such features are extracted directly from each nucleus to provide discriminative characteristics of different stained nuclei. The output generated from the first and second quantitative measures are used collectively to calculate the percentage of positive nuclei (PS). The third measure proposes a novel automated method for determining the staining intensity level of positive nuclei or what is known as the intensity score (IS). The minor intensity features are observed and used to classify low, intermediate and high stained positive nuclei. Statistical methods were applied throughout the research to validate the system results against the ground truth pathology data. Experimental results demonstrate the effectiveness of the proposed approach and provide high accuracy when compared to the ground truth pathology data

Characterising LINC complex roles in 3D epithelial migration and breast cancer metastasis

Cell migration is essential for the development of multicellular organisms; with disruptions in this process contributing to diseases such as cancer, neurological disorders and musculoskeletal diseases. The LINC (Linker of Nucleoskeleton and Cytoskeleton) complex is an evolutionary conserved proteinaceous structure, critical for maintaining proper cellular migration. This multifunctional complex provides a physical connection between the nuclear interior and the cytoskeleton, with disruptions stimulating loss of directed cell migration, compromised nuclear structure and abnormal cellular signalling. As it is also noted that the nucleus in many cells is the stiffest cellular component, it is suggested that LINC complex disruptions may be key in increasing the migration potential of cells through both in vivo and in vitro 3D environments. This project aimed to investigate the roles of LINC complex disruptions on keratinocyte morphological and migratory behaviours in 2D, and both non-restrictive and space-restrictive 3D culture environments, through the application of dominant negative SUN1 mutants. Through extensive analysis, it was identified that these mutants exhibited altered cell-cell and cell-substratum attachment phenotypes, alongside increased nuclear heights. It was also demonstrated that the LINC disrupted mutants displayed a migration advantage in space-restricted 3D environments, which was attributed to a decrease in nuclear stiffness. Through fibroblast incorporation to the 3D scaffolds used, it was further shown that LINC disrupted keratinocytes displayed increased levels of differentiation markers, alongside increased cellular stacking phenotypes across scaffolds surface regions, potentially attributed to alterations in Hippo pathway signalling. The migratory phenotypes observed in DN mutants closely resemble that of high-grade cancer cells, able to migrate through space-restrictive environments during metastasis. Comprehensive protein expression and localisation analysis across a range of breast cancer cell lines and tissues suggested that several LINC complex components display altered expression levels closely linked to cancer progression, most significantly a down-regulation of lower nesprin-1/-2 isoforms was identified. As following investigations later suggested Nup88 as an upstream regulator of nesprin-2, able to bind C-terminal regions, it’s suggested that these phenotypes link closely to that observed in high-grade cancers. Together, the data presented suggests that LINC complex disruptions increase migration potential of cells through restrictive 3D environments due to a decrease in nuclear stiffness, comparable to that observed across high-grade breast cancer cell lines

Glaucoma

This book addresses the basic and clinical science of glaucomas, a group of diseases that affect the optic nerve and visual fields and is usually accompanied by increased intraocular pressure. The book incorporates the latest development as well as future perspectives in glaucoma, since it has expedited publication. It is aimed for specialists in glaucoma, researchers, general ophthalmologists and trainees to increase knowledge and encourage further progress in understanding and managing these complicated diseases

Silicon Nanodevices

This book is a collection of scientific articles which brings research in Si nanodevices, device processing, and materials. The content is oriented to optoelectronics with a core in electronics and photonics. The issue of current technology developments in the nanodevices towards 3D integration and an emerging of the electronics and photonics as an ultimate goal in nanotechnology in the future is presented. The book contains a few review articles to update the knowledge in Si-based devices and followed by processing of advanced nano-scale transistors. Furthermore, material growth and manufacturing of several types of devices are presented. The subjects are carefully chosen to critically cover the scientific issues for scientists and doctoral students

Non-covalent interactions in organotin(IV) derivatives of 5,7-ditertbutyl- and 5,7-diphenyl-1,2,4-triazolo[1,5-a]pyrimidine as recognition motifs in crystalline self- assembly and their in vitro antistaphylococcal activity

Non-covalent interactions are known to play a key role in biological compounds due to their stabilization of the tertiary and quaternary structure of proteins [1]. Ligands similar to purine rings, such as triazolo pyrimidine ones, are very versatile in their interactions with metals and can act as model systems for natural bio-inorganic compounds [2]. A considerable series (twelve novel compounds are reported) of 5,7-ditertbutyl-1,2,4-triazolo[1,5-a]pyrimidine (dbtp) and 5,7-diphenyl- 1,2,4-triazolo[1,5-a]pyrimidine (dptp) were synthesized and investigated by FT-IR and 119Sn M\uf6ssbauer in the solid state and by 1H and 13C NMR spectroscopy, in solution [3]. The X-ray crystal and molecular structures of Et2SnCl2(dbtp)2 and Ph2SnCl2(EtOH)2(dptp)2 were described, in this latter pyrimidine molecules are not directly bound to the metal center but strictly H-bonded, through N(3), to the -OH group of the ethanol moieties. The network of hydrogen bonding and aromatic interactions involving pyrimidine and phenyl rings in both complexes drives their self-assembly. Noncovalent interactions involving aromatic rings are key processes in both chemical and biological recognition, contributing to overall complex stability and forming recognition motifs. It is noteworthy that in Ph2SnCl2(EtOH)2(dptp)2 \u3c0\u2013\u3c0 stacking interactions between pairs of antiparallel triazolopyrimidine rings mimick basepair interactions physiologically occurring in DNA (Fig.1). M\uf6ssbauer spectra suggest for Et2SnCl2(dbtp)2 a distorted octahedral structure, with C-Sn-C bond angles lower than 180\ub0. The estimated angle for Et2SnCl2(dbtp)2 is virtually identical to that determined by X-ray diffraction. Ph2SnCl2(EtOH)2(dptp)2 is characterized by an essentially linear C-Sn-C fragment according to the X-ray all-trans structure. The compounds were screened for their in vitro antibacterial activity on a group of reference staphylococcal strains susceptible or resistant to methicillin and against two reference Gramnegative pathogens [4] . We tested the biological activity of all the specimen against a group of staphylococcal reference strains (S. aureus ATCC 25923, S. aureus ATCC 29213, methicillin resistant S. aureus 43866 and S. epidermidis RP62A) along with Gram-negative pathogens (P. aeruginosa ATCC9027 and E. coli ATCC25922). Ph2SnCl2(EtOH)2(dptp)2 showed good antibacterial activity with a MIC value of 5 \u3bcg mL-1 against S. aureus ATCC29213 and also resulted active against methicillin resistant S. epidermidis RP62A

Flexible Synapse Detection in Fluorescence Micrographs by Modeling Human Expert Grading

Herold J, Friedenberger M, Bode M, Rajpoot N, Schubert W, Nattkemper TW. Flexible Synapse Detection in Fluorescence Micrographs by Modeling Human Expert Grading. In: Proc. of 2008 IEEE International Symposium on Biomedical Imaging. 2008.A particularly difficult task in molecular imaging is the analysis of fluorescence microscopy images of neural tissue, as they usually exhibit a high density of objects with diffuse signals. To automate synapse detection in such images, one has to simulate aspects of human pattern recognition skills to account for low signal-to-noiseratios. We propose a machine learning based method that allows a direct integration of the experts’ visual expertise who tag a low number of referential synapses according to their degree of synapse likeness. The sensitivity and positive predictive values show that by using graded likeness information in our learning algorithm we can provide an intuitively tunable tool for neural tissue slide evaluation

In vitro expanded human CD4+CD25+ regulatory T cells suppress effector T cell proliferation.

Regulatory T cells (Tregs) have been shown to be critical in the balance between autoimmunity and tolerance and have been implicated in several human autoimmune diseases. However, the small number of Tregs in peripheral blood limits their therapeutic potential. Therefore, we developed a protocol that would allow for the expansion of Tregs while retaining their suppressive activity. We isolated CD4+CD25 hi cells from human peripheral blood and expanded them in vitro in the presence of anti-CD3 and anti-CD28 magnetic Xcyte Dynabeads and high concentrations of exogenous Interleukin (IL)-2. Tregs were effectively expanded up to 200-fold while maintaining surface expression of CD25 and other markers of Tregs: CD62L, HLA-DR, CCR6, and FOXP3. The expanded Tregs suppressed proliferation and cytokine secretion of responder PBMCs in co-cultures stimulated with anti-CD3 or alloantigen. Treg expansion is a critical first step before consideration of Tregs as a therapeutic intervention in patients with autoimmune or graft-versus-host disease