Physiological Hypoxia (Physioxia) Impairs the Early Adhesion of Single Lymphoma Cell to Marrow Stromal Cell and Extracellular Matrix. Optical Tweezers Study.

Adhesion is critical for the maintenance of cellular structures as well as intercellular communication, and its dysfunction occurs prevalently during cancer progression. Recently, a growing number of studies indicated the ability of oxygen to regulate adhesion molecules expression, however, the influence of physiological hypoxia (physioxia) on cell adhesion remains elusive. Thus, here we aimed: (i) to develop an optical tweezers based assay to precisely evaluate single diffuse large B-cell lymphoma (DLBCL) cell adhesion to neighbor cells (mesenchymal stromal cells) and extracellular matrix (Matrigel) under normoxia and physioxia; and, (ii) to explore the role of integrins in adhesion of single lymphoma cell. We identified the pronouncedly reduced adhesive properties of lymphoma cell lines and primary lymphocytes B under physioxia to both stromal cells and Matrigel. Corresponding effects were shown in bulk adhesion assays. Then we emphasized that impaired ß1, ß2 integrins, and cadherin-2 expression, studied by confocal microscopy, account for reduction in lymphocyte adhesion in physioxia. Additionally, the blockade studies conducted with anti-integrin antibodies have revealed the critical role of integrins in lymphoma adhesion. To summarize, the presented approach allows for precise confirmation of the changes in single cell adhesion properties provoked by physiological hypoxia. Thus, our findings reveal an unprecedented role of using physiologically relevant oxygen conditioning and single cell adhesion approaches when investigating tumor adhesion in vitro.

[Mondor's disease or systemic sclerosis - diagnostic and therapeutic dilemmas].

In this article we have presented a case of 31-year-old male with thickening of the skin and tender cord of superficial veins of the penis and laboratory findings of the high titer of PM/Scl-100 antibodies in the serum. The patient was referred to the Rheumatology Department due to suspected systemic sclerosis. The HRCT scan of his lungs revealed fibrous degeneration and ground-glass opacity in the lower lobes. Capillaroscopy showed abnormalities, which were not typical for systemic sclerosis. In Doppler ultrasound examination of penis, superficial dorsal and circumflex veins thrombosis and inflammatory infiltration were observed. Taking into account the entire clinical picture, the patient was diagnosed with rare penile Mondor's disease and was under surveillance for systemic sclerosis.

Significance of Matrix Metalloproteinase 9 Expression as Supporting Marker to Cytokeratin 19 mRNA in Sentinel Lymph Nodes in Breast Cancer Patients.

One-step nucleic acid amplification (OSNA) detects and quantifies, with the use of a polymerase chain reaction, the presence of cytokeratin 19 mRNA in sentinel lymph nodes. The main advantage of the OSNA assay is the avoidance of second surgery in case of positive sentinel lymph node diagnosis. The objective of this study was to evaluate the significance of matrix metalloproteinase 9 expression by immunohistochemistry as supporting marker to cytokeratin 19 mRNA in sentinel lymph nodes in breast cancer patients and to relate this expression with clinicopathological data. This study was conducted on fresh sentinel lymph nodes obtained from 40 patients with tumors classified as carcinoma of no special type. The presence of metastatic cells in the slices of lymph nodes was evaluated by immunohistochemistry using antibodies for CK19 and MMP-9. Expression of CK19 and MMP-9 in lymph nodes was also confirmed by means of Western blot analysis. Results indicated that the strongest correlation with CK19 mRNA was displayed by MMP-9, CK19 (by immunohistochemistry, IHC), and nodal metastases (p < 0.001). Higher histological grading also positively correlated with CK19 mRNA, however that correlation was less significant. Since MMP-9 shows very strong correlation with CK19 mRNA in breast carcinoma of no special type metastases, expression of MMP-9 in sentinel lymph nodes should be considered as useful method whenever OSNA analysis is not available.

Mechanical Properties of Sugar Beet Roots under Impact Loading Conditions.

Root damages due to mechanical impacts result in deterioration in commercial sugar beet quality. In order to determine the mechanical characteristics of roots, a stand equipped with a pendulum enabling impact investigations of whole beets was used. The roots were stored in a monitored environment for up to 5 days (temperature 15 ± 2 °C, 40 ± 2%). During the tests, the beets were struck against a flat steel resistant surface with the velocities Vimp = 0.5, 1.0 and 1.5 m·s-1. The measurements of local root curvatures in three chosen impact areas and the deformation (dmax) allowed modelling of the volume of contact (CV) by means of the ellipsoid cap. These investigations enabled the determination of the relations between the maximal impact force, Fmax, the impact energy, Eimp, and the absorbed energy, Eabs, as well as the contact volume and impact velocity, taking into account the root storage time, St. It was found that the maximal impact force increased with increasing impact velocity and decreased with the storage time for each group of roots. With increasing velocity, there were also increases in the following: impact energy, absorbed energy, contact volume and maximal deformation, as well as absorbed energy, referred to as the mass Eabs-v from Vimp. The mean values of the stresses (σmax), being the quotients of the impact force (Fmax) and the surface area of the ellipsoid cap base (ABE), were 0.81-1.17 MPa, 1.064-1.59 MPa and 1.45-1.77 MPa for the velocities of 0.5, 1.0 and 1.5 m·s-1, respectively. It was confirmed that the statistical significance of the mentioned parameters changes depending on the impact velocity.

Combined small cell and squamous cell carcinoma of the larynx.

A case of combined small cell and squamous cell carcinoma of the larynx in a male patient in the sixth decade of life is reported. The etiopathogenesis of this combined tumor remains unclear; however, a number of hypotheses were proposed in the past including the pivotal role of Kulchitsky, squamous cells and the glandular cells. The gene mutations may also play an important role in laryngeal carcinogenesis. This unusual type of laryngeal combined carcinoma has previously been reported worldwide in only 17 cases. This is an extremely rare tumor the histological nature of which makes the diagnosis more complicated than in other types of laryngeal cancers. The diagnosis of this carcinoma is based on light microscopy and should be supported by immunohistochemical studies. In our case, the tumor was growing in the left pyriform sinus. Metastatic neck lymph nodes were found on the left side, but no distant metastases were observed. Microscopic sections revealed a combined tumor composed of small cell carcinoma neuroendocrine type and non-keratinizing squamous cell carcinoma. Positive reaction to p16, bcl-2, thyroid transcription factor 1, synaptophysin and chromogranin A in the small cell neuroendocrine type carcinoma component was observed. The cells from squamous cell carcinoma component showed positive reaction to p63, high-molecular-weight cytokeratin and cytokeratin 5/6.

Mechanical Characteristics of the Flebogrif System-The New System of Mechano-Chemical Endovenous Ablation.

Non-thermal endovenous ablations, due to the lowest probability of complications, are the new method of treating chronic venous insufficiency-one of the most common diseases globally. The Flebogrif system (Balton Sp. z o.o., Warsaw, Poland) is a new mechano-chemical ablation system causing the mechanical damage of endothelium that allows for better sclerosant penetration into its wall. The purpose of the article is to provide mechanical characteristics in the form of force-displacement dependence for a single cutting element, and a bundle of cutting elements of Flebogrif as a whole for different levels of protrusion of the bundle of cutting elements. A TA.HD plus (Stable Micro Systems, Godalming, UK) analyzer equipped with special handles, was used for characteristics testing. The head movement speed used was 5 mm·s-1. The Flebogrif system was tested for three cutting element protrusion levels: L = Lmax, L = 0.9·Lmax, and L = 0.8·Lmax. Before testing, geometric measurement of the spacing of the cutting elements for three proposed protrusions was performed. It was established that decreasing the working length of the cutting elements will increase their rigidity, and, as a result, increase the force exerted on the internal surface of the vein wall. The obtained characteristics will allow for specifying contact force variability ranges and the corresponding diameter ranges of operated veins.

Effect of Nesfatin-1 on Rat Humerus Mechanical Properties under Quasi-Static and Impact Loading Conditions.

The investigations on the response of bone tissue under different loading conditions are important from clinical and engineering points of view. In this paper, the influence of nesfatin-1 administration on rat humerus mechanical properties was analyzed. The classical three-point bending and impact tests were carried out for three rat bone groups: control (SHO), the humerus of animals under the conditions of established osteopenia (OVX), and bones of rats receiving nesfatin-1 after ovariectomy (NES). The experiments proved that the bone strength parameters measured under various mechanical loading conditions increased after the nesfatin-1 administration. The OVX bones were most susceptible to deformation and had the smallest fracture toughness. The SEM images of humerus fracture surface in this group showed that ovariectomized rats had a much looser bone structure compared to the SHO and NES females. Loosening of the bone structure was also confirmed by the densitometric and qualitative EDS analysis, showing a decrease in the OVX bones' mineral content. The samples of the NES group were characterized by the largest values of maximum force obtained under both quasi-static and impact conditions. The energies absorbed during the impact and the critical energy for fracture (from the three-point bending test) were similar for the SHO and NES groups. Statistically significant differences were observed between the mean Fi max values of all analyzed sample groups. The obtained results suggest that the impact test was more sensitive than the classical quasi-static three-point bending one. Hence, Fi max could be used as a parameter to predict bone fracture toughness.

Large-Scale Proteomic Analysis of Follicular Lymphoma Reveals Extensive Remodeling of Cell Adhesion Pathway and Identifies Hub Proteins Related to the Lymphomagenesis.

Follicular lymphoma (FL) represents the major subtype of indolent B-cell non-Hodgkin lymphomas (B-NHLs) and results from the malignant transformation of mature B-cells in lymphoid organs. Although gene expression and genomic studies have identified multiple disease driving gene aberrations, only a few proteomic studies focused on the protein level. The present work aimed to examine the proteomic profiles of follicular lymphoma vs. normal B-cells obtained by fine-needle aspiration biopsy (FNAB) to gain deep insight into the most perturbed pathway of FL. The cells of interest were purified by magnetic-activated cell sorting (MACS). High-throughput proteomic profiling was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and allowed to identify of 6724 proteins in at least 75% of each group of samples. The 'Total Protein Approach' (TPA) was applied to the absolute quantification of proteins in this study. We identified 1186 differentially abundant proteins (DAPs) between FL and control samples, causing an extensive remodeling of several molecular pathways, including the B-cell receptor signaling pathway, cellular adhesion molecules, and PPAR pathway. Additionally, the construction of protein-protein interactions networks (PPINs) and identification of hub proteins allowed us to indicate the key player proteins for FL pathology. Finally, ICAM1, CD9, and CD79B protein expression was validated in an independent cohort by flow cytometry (FCM), and the results were consistent with the mass spectrometry (MS) data.

Differentiation of single lymphoma primary cells and normal B-cells based on their adhesion to mesenchymal stromal cells in optical tweezers.

We have adapted a non-invasive method based on optical tweezers technology to differentiate between the normal B-cells and the B-cell non-Hodgkin lymphoma (B-NHL) cells derived from clinical samples. Our approach bases on the nascent adhesion between an individual B-cell and a mesenchymal stromal cell. In this study, a single B-cell was trapped and optically seeded on a mesenchymal stromal cell and kept in a direct contact with it until a stable connection between the cells was formed in time scale. This approach allowed us to avoid the introduction of any exogenous beads or chemicals into the experimental setup which would have affected the cell-to-cell adhesion. Here, we have provided new evidence that aberrant adhesive properties found in transformed B-cells are related to malignant neoplasia. We have demonstrated that the mean time required for establishing adhesive interactions between an individual normal B-cell and a mesenchymal stromal cell was 26.7 ± 16.6 s, while for lymphoma cell it was 208.8 ± 102.3 s, p < 0.001. The contact time for adhesion to occur ranged from 5 to 90 s and from 60 to 480 s for normal B-cells and lymphoma cells, respectively. This method for optically controlled cell-to-cell adhesion in time scale is beneficial to the successful differentiation of pathological cells from normal B-cells within the fine needle aspiration biopsy of a clinical sample. Additionally, variations in time-dependent adhesion among subtypes of B-NHL, established here by the optical trapping, confirm earlier results pertaining to cell heterogeneity.

Pattern of Melanotransferrin Expression in Human Colorectal Tissues: An Immunohistochemical Study on Potential Clinical Application.

BACKGROUND: Our previous liquid chromatography-tandem mass spectrometry (LC-MS/MS) study on colorectal cancer proteome resulted in identification of 10,000 differentially expressed proteins. We observed a significantly changed expression of 25% of all identified proteins between patient and matched adjacent normal mucosa, carcinoma and colorectal adenoma, including melanotransferrin. Herein, we consider this protein as a potential biomarker of colorectal cancer. MATERIALS AND METHODS: Immunohistochemical detection of melanotransferrin was carried-out to localize its expression pattern within the colorectal tissues by tissue microarray. The diagnostic utility of melanotransferrin was evaluated in patient serum by enzyme-linked immunosorbent assay (ELISA). RESULTS: Strong melanotransferrin expression was found to be related to clinicopathological characteristics, lymph node involvement (p=0.008), tumor localization in colon (p=0.001), presence of mucin (p<0.013) and increasing tumor grade (p<0.001). Melanotransferrin level in serum from patients with colorectal cancer was significantly higher than that in healthy controls (p<0.001). CONCLUSION: We provide novel evidence that melanotransferrin may be involved in transformation from benign tumor to malignancy and is a marker of an invasive tumor phenotype.

Immunohistochemical study of nuclear ubiquitous casein and cyclin-dependent kinase substrate 1 in invasive breast carcinoma of no special type.

The aim of the present study was to investigate the immunohistochemical expression of nuclear ubiquitous casein and cyclin-dependent kinases substrate 1 (NUCKS1) in invasive breast carcinoma of no special type, in association with clinicopathological characteristics, including the tumor grade, frequency of lymph node involvement and distant metastasis. In addition, associations between NUCKS1 and other tumor subtype markers, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), Ki-67 and cytokeratin 5/6 (CK 5/6), were investigated. NUCKS1 expression was shown to be associated with the formation of distant metastases and lymph node involvement. Furthermore, an association between the presence of NUCKS1 and histological grading was observed. The results confirmed that the expression of NUCKS1 in low grade invasive breast carcinoma of no special type was significantly less common compared with cases of high grade carcinoma. With regard to the additional tumor subtype markers, NUCKS1 expression was demonstrated to be significantly associated with Ki-67 and CK 5/6; however, no association was identified with ER, PR and HER2. Therefore, NUCKS1 may be a novel prognostic marker in the histopathological evaluation of invasive breast carcinoma of no special type.