A brief history of angiogenesis assays

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Paul Langerhans: hodočasnik koji je "putovao" of funkcionalne histologije do biologije mora

The nineteenth century was the time of a real revolution in science and medicine. A lot of seminal discoveries in medicine and biology were done in this time, and many of them were coincident with the introduction of the compound microscope by Hermann van Deijl and the standard histological technique by Paul Ehrlich. The main tissue types and individual cells were characterized and originally classified more than hundred years ago, although less attention was paid to their basic functions. This was mainly due to the modality of tissue specimen processing that allowed particularly detailed descriptive studies. Even so, we can notice some attempts to correlate the structure with the function. The German scientist Paul Langerhans, well-known for the discovery of Langerhans islets of the pancreas and Langerhans cells from the epidermis, tried to change the conventional fate of morphological studies introducing in his works functional hypothesis based on traditional microscopic observations even from the beginning of his scientific career. Paul Langerhans was a complex personality of the second half of the nineteenth century, not only in medicine, but also in other fields of biology. In the present review, presented is the life and research activity of Paul Langerhans, not only because of the importance of his discoveries, but also for perspectives that were opened by these findings in unexpected fields of medicine and biology.Devetnaesto stoljeće bilo je vrijeme prave revolucije u znanosti i medicini. U tom je stoljeću došlo do mnogih otkrića u medicini i biologiji, a mnoga od njih su se podudarala s primjenom mikroskopa (Hermann van Deijl) i uvođenjem standardne histološke tehnike (Paul Ehrlich). Glavne vrste tkiva i pojedinačne stanice karakterizirane su i izvorno klasificirane prije više od stotinu godina, iako je manje pozornosti bilo posvećeno njihovim osnovnim funkcijama. To je uglavnom bilo zbog modaliteta obrade uzoraka tkiva koja je omogućavala posebno detaljne deskriptivne studije. Ipak, primjećuju se neki pokušaji povezivanja strukture s funkcijom. Njemački znanstvenik Paul Langerhans, poznat po otkriću Langerhansovih otočića gušterače i Langerhansovih stanica u epidermisu, pokušao je promijeniti konvencionalnu sudbinu morfoloških istraživanja uvodeći već od početka svoje znanstvene karijere u svoja istraživanja funkcionalnu hipotezu koja se temelji na tradicionalnim mikroskopskim promatranjima. Paul Langerhans bio je složena ličnost druge polovice XIX. stoljeća ne samo u medicini nego i u drugim područjima biologije. U ovom radu predstavljen je njegov život i njegova istraživačka djelatnost ne samo zbog važnosti njegovih otkrića već i zbog gledišta koja su ta otkrića potaknula u neočekivanim područjima medicine i biologije

B-cell lymphoma-2 receptor in human primary breast and its lymph node metastases: more than a surrogate marker

Background: Due to its anti-apoptotic and anti-proliferative contradictory functions, BCL2 role in breast carcinoma progression is not clearly understood. The purpose of this study was to highlight BCL2 expression during metastatic progression of invasive breast carcinoma of no special type (NST). Materials and methods: The specimens, primary tumors and corresponding lymph node metastases (LNM) from 84 patients were immunostained for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor (HER)-2, basal cytokeratin CK5, nuclear protein Ki67 and B-cell lymphoma (Bcl)-2 receptor. Results: BCL2 expression was higher at primary site than in axillary metastases. Its score correlates positively with hormone receptors’ level and negatively with HER2, CK5 and Ki67 at both sites. Switch of molecular profile was determined in 22.62% of cases. BCL2 expression was not influenced by subtypes switch. Changes of BCL2 expression were found in 25% of cases with stable molecular subtype. The Luminal A and Luminal B/Ki67 were encountered in the majority of BCL2 transitions, mainly from positive to negative state. Conclusions: Molecular subtypes and BCL2 expression are not stable during tumor progression and metastatic development. In the present study we established immunohistochemically that BCL2 is not influenced by subtypes’ transitions. BCL2 switches were encountered only in cases with a stable HER2, Luminal A or B phenotypes. We expect a further confirmation of our results by other research groups. Key words: BCL2, breast carcinoma, immunohistochemistry, molecular subtypes, metastases

Oddziaływanie kwaśnego białka włókienkowego gleju i białka S100 w gruczolakach przysadki mózgowej: dwóch czy więcej graczy?

Introduction: S100 protein and GFAP expression in pituitary adenomas tumour cells is not well known; few correlations with other prognostic or therapeutic factors have previously been reported in pituitary adenomas. We aim to elucidate their involvement in the pathogenesis of pituitary adenomas and to establish the correlation of their expression with different growth factors and growth factor receptors known to have a prognostic and/or therapeutic role. Material and methods: Sixty-one cases of pituitary adenomas were immunohistochemically assessed for the expression of GFAP and S100 protein in both tumour cells and FS cells, in close relationship with hormone profile, and correlated with vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) expression, previously studied by our team. Results: GFAP and S100 protein were expressed both in tumour cells and FS cells. Differences between morphology, distribution, and density of GFAP+ FS cells and S100+ FS cells were observed according to the hormone profile of pituitary adenomas. GFAP and S100 protein expression in tumour cells was significantly related to hormone profile of pituitary adenomas and also with VEGF and EGFR expression. Conclusions: GFAP and S100 protein expressions in tumour cells from pituitary adenomas are influenced by hormone profile. Our re­sults support the presence of two molecular subtypes of FS cells GFAP+/VEGF+/S100 respectively and another one that is GFAP-/S100+/EGFR+ simultaneously with the classical variant GFAP+/S100+. It is possible that S100+/EGFR+ pituitary adenomas represent a group of pituitary adenomas with an aggressive behaviour and a high ability of invasion and recurrence.Wstęp: Ekspresja białka S100 i kwaśnego białka włókienkowego gleju (GFAP, glial fibrillary acid protein) w gruczolakach przysadki mózgowej nie została dobrze poznana. Dostępne są nieliczne publikacje dotyczące korelacji tych cząsteczek z innymi czynnikami prognostycznymi lub terapeutycznymi w gruczolakach przysadki. Celem niniejszej pracy jest wyjaśnienie udziału tych białek w patogenezie gruczolaków przysadki i ustalenie korelacji między ich ekspresją a różnymi czynnikami wzrostu lub receptorami czynników wzrostu o znanej wartości prognostycznej i/lub terapeutycznej. Materiał i metody: Sześćdziesiąt sześć przypadków gruczolaka przysadki zbadano metodami immunohistochemicznymi w celu oceny ekspresji białek GFAP i S100 zarówno w komórkach guza, jak i w komórkach pęcherzykowo-gwiaździstych (FS, folliculo-stellate cells) oraz przeanalizowania uzyskanych wyników w ścisłej zależności z profilami hormonalnymi gruczolaków i w korelacji z ekspresją czynnika wzrostu śródbłonka naczyniowego (VEGF, vascular endothelial growth factor) i receptora naskórkowego czynnika wzrostu (EGFR, epidermal growth factor receptor), ocenianych w badaniu przeprowadzonym wcześniej przez nasz zespół. Wyniki: Ekspresję białek GFAP i S100 stwierdzono zarówno w komórkach guza, jak i w komórkach FS. Zaobserwowano różnice pod względem morfologii, rozkładu i gęstości komórek FS z dodatnią ekspresją białek GFAP i S100 (GFAP+FS i S100+FS) odpowiadające profilowi hormonalnemu gruczolaków przysadki. Ekspresja białek GFAP i S100 w komórkach guza była istotnie związana z profilem hormonalnym gruczolaków przysadki, a także z ekspresją VEGF i EGFR. Wnioski: Ekspresja białek GFAP i S100 w komórkach gruczolaka przysadki zależy od profilu hormonalnego guza. Uzyskane w badaniu wyniki potwierdzają obecność dwóch podtypów molekularnych komórek FS GFAP+/VEGF+/S100 oraz jednego typu GFAP–/S100+/EGFR+ występujących jednocześnie z klasycznym wariantem GFAP+/S100+. Możliwe, że gruczolaki przysadki z ekspresją S100+/EGFR+ należą do grupy tych nowotworów cechujących się agresywnością i dużą zdolnością do naciekania i nawrotów

Toward a Molecular Classification of the Head and Neck Squamous Cell Carcinoma

The frequency of the squamous cell carcinoma of the head and neck is constantly increasing, with over 550.000 new cases registered globally each year. The conventional histopathological diagnosis most commonly indicates the squamous cell carcinoma as tumor type and G2 as differentiation grade. Despite of this relative morphological uniformity, there is a great heterogeneity in the molecular profile, the therapeutic response and prognosis. Most probably, this entity includes many diseases, similar in basic morphologic features, but different in the biological behavior. Trying to answer this question and to show discrepancies when they exist, we have evaluated in this book chapter, our own results and data from the literature in terms of molecular profile at the protein level, including the spectrum of proliferation markers, growth factors and their receptors, stromal proliferation, angiogenesis and lymphangiogenesis. These data will allow to identify some major criteria for a better stratification of cases, selected for gene analysis and personalized therapy as a future perspective and direction

Graphene nanoplatelets-sericin surface-modified Gum alloy for improved biological response

In this study a “Gum Metal” titanium-based alloy, Ti-31.7Nb-6.21Zr-1.4Fe-0.16O, was synthesized by melting and characterized in order to evaluate its potential for biomedical applications. The results showed that the newly developed alloy presents a very high strength, high plasticity and a low Young\u27s modulus relative to titanium alloys currently used in medicine. For further bone implant applications, the newly synthesized alloy was surface modified with graphene nanoplatelets (GNP), sericin (SS) and graphene nanoplatelets/sericine (GNP–SS) composite films via Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. The characterization of each specimen was monitored by scanning electron microscopy (SEM), atomic force microscopy (AFM), contact angle (CA) measurements, and Fourier Transform Infrared Spectroscopy (FTIR). The materials\u27 surface analyses suggested the successful coating of GNP, SS and GNP–SS onto the alloy surface. Additionally, the activities of pre-osteoblasts such as cell adhesion, cytoskeleton organization, cell proliferation and differentiation potentials exhibited on these substrates were investigated. Results showed that the GNP–SS-coated substrate significantly enhanced the growth and osteogenic differentiation of MC3T3-E1 cells when compared to bare and GNP-coated alloy. Collectively, the results show that GNP–SS surface-modified Gum alloy can modulate the bioactivity of the pre-osteoblasts holding promise for improved biological response in vivo

Deconvolving the contributions of cell-type heterogeneity on cortical gene expression

Complexity of cell-type composition has created much skepticism surrounding the interpretation of bulk tissue transcriptomic studies. Recent studies have shown that deconvolution algorithms can be applied to computationally estimate cell-type proportions from gene expression data of bulk blood samples, but their performance when applied to brain tissue is unclear. Here, we have generated an immunohistochemistry (IHC) dataset for five major cell-types from brain tissue of 70 individuals, who also have bulk cortical gene expression data. With the IHC data as the benchmark, this resource enables quantitative assessment of deconvolution algorithms for brain tissue. We apply existing deconvolution algorithms to brain tissue by using marker sets derived from human brain single cell and cell-sorted RNA-seq data. We show that these algorithms can indeed produce informative estimates of constituent cell-type proportions. In fact, neuronal subpopulations can also be estimated from bulk brain tissue samples. Further, we show that including the cell-type proportion estimates as confounding factors is important for reducing false associations between Alzheimer\u27s disease phenotypes and gene expression. Lastly, we demonstrate that using more accurate marker sets can substantially improve statistical power in detecting cell-type specific expression quantitative trait loci (eQTLs)

Trans-pQTL study identifies immune crosstalk between Parkinson and Alzheimer loci

Objective: Given evidence from genetic studies, we hypothesized that there may be a shared component to the role of myeloid function in Parkinson and Alzheimer disease (PD and AD) and assessed whether PD susceptibility variants influenced protein expression of well-established AD-associated myeloid genes in human monocytes. Methods: We repurposed data in which AD-related myeloid proteins CD33, TREM1, TREM2, TREML2, TYROBP, and PTK2B were measured by flow cytometry in monocytes from 176 participants of the PhenoGenetic Project (PGP) and Harvard Aging Brain Study. Linear regression was used to identify associations between 24 PD risk variants and protein expression. The 2 cohorts were meta-analyzed in a discovery analysis, and the 4 most strongly suggestive results were validated in an independent cohort of 50 PGP participants. Results: We discovered and validated an association between the PD risk allele rs12456492G in the RIT2 locus and increased CD33 expression (pjoint = 3.50 × 10−5) and found strongly suggestive evidence that rs11060180A in the CCDC62/HIP1R locus decreased PTK2B expression (pjoint = 1.12 × 10−4). Furthermore, in older individuals, increased CD33 expression on peripheral monocytes was associated with a greater burden of parkinsonism (p = 0.047), particularly bradykinesia (p = 6.64 × 10−3). Conclusions: We find that the rs12456492 PD risk variant affects expression of AD-associated protein CD33 in peripheral monocytes, which suggests that genetic factors for these 2 diseases may converge to influence overlapping innate immune-mediated mechanisms that contribute to neurodegeneration. Furthermore, the effect of the rs12456492G PD risk allele on increased CD33 suggests that the inhibition of certain myeloid functions may contribute to PD susceptibility, as is the case for AD

Identification of Stage-Specific Breast Markers using Quantitative Proteomics

YesMatched healthy and diseased tissues from breast cancer patients were analyzed by quantitative proteomics. By comparing proteomic profiles of fibroadenoma (benign tumors, three patients), DCIS (noninvasive cancer, three patients), and invasive ductal carcinoma (four patients), we identified protein alterations that correlated with breast cancer progression. Three 8-plex iTRAQ experiments generated an average of 826 protein identifications, of which 402 were common. After excluding those originating from blood, 59 proteins were significantly changed in tumor compared with normal tissues, with the majority associated with invasive carcinomas. Bioinformatics analysis identified relationships between proteins in this subset including roles in redox regulation, lipid transport, protein folding, and proteasomal degradation, with a substantial number increased in expression due to Myc oncogene activation. Three target proteins, cofilin-1 and p23 (increased in invasive carcinoma) and membrane copper amine oxidase 3 (decreased in invasive carcinoma), were subjected to further validation. All three were observed in phenotype-specific breast cancer cell lines, normal (nontransformed) breast cell lines, and primary breast epithelial cells by Western blotting, but only cofilin-1 and p23 were detected by multiple reaction monitoring mass spectrometry analysis. All three proteins were detected by both analytical approaches in matched tissue biopsies emulating the response observed with proteomics analysis. Tissue microarray analysis (361 patients) indicated cofilin-1 staining positively correlating with tumor grade and p23 staining with ER positive status; both therefore merit further investigation as potential biomarkers.Cyprus Research Promotion Foundation, Yorkshire Cancer Researc