Adcitmer® , a new CD56-targeting monomethyl auristatin E-conjugated antibody, is a potential therapeutic approach in Merkel cell carcinoma.

BACKGROUND: Merkel cell carcinoma (MCC) is an aggressive skin cancer, whose tumour cells often express CD56. While immune checkpoint inhibitors constitute a major advance for treating patients with MCC with advanced disease, new therapeutic options are still urgently required. OBJECTIVES: To produce and evaluate the therapeutic performance of a new antibody-drug conjugate (Adcitmer® ) targeting CD56 in preclinical models of MCC. METHODS: CD56 expression was evaluated in a MCC cohort (immunohistochemistry on a tissue microarray of 90 tumour samples) and MCC cell lines. Interaction of an unconjugated CD56-targeting antibody with CD56+ MCC cell lines was investigated by immunohistochemistry and imaging flow cytometry. Adcitmer® product was generated by the bioconjugation of CD56-targeting antibody to a cytotoxic drug (monomethyl auristatin E) using the McSAF Inside® bioconjugation process. The chemical properties and homogeneity of Adcitmer® were characterized by hydrophobic interaction chromatography. Adcitmer® cytotoxicity was evaluated in vitro and in an MCC xenograft mice model. RESULTS: Similar to previous reports, CD56 was expressed by 66% of MCC tumours in our cohort, confirming its relevance as a therapeutic target. Specific binding and internalization of the unconjugated CD56-targeting antibody was validated in MCC cell lines. The high homogeneity of the newly generated Adcitmer® was confirmed by hydrophobic interaction chromatography. The CD56-mediated cytotoxicity of Adcitmer® was demonstrated in vitro in MCC cell lines. Moreover, Adcitmer® significantly reduced tumour growth in a MCC mouse model. CONCLUSIONS: Our study suggests that Adcitmer® should be further assessed as a therapeutic option in patients with MCC, as an alternative therapy or combined with immune checkpoint inhibitors.

[Determining the level of occurrence of the lipid nucleus of atherosclerotic plaques in the abdominal aorta by means of infrared Fourier spectroscopy].

The article deals with the problem of diagnosis of atherosclerotic alterations in abdominal aorta wall in order to reduce atherosclerosis-related mortality. Targeted treatment requires determining the exact mechanism of development of atherosclerosis. Infrared Fourier spectroscopy is suggested as the most rapid and accurate method of diagnosis and analysis of the content of various organic compounds playing a particular role in the development of the atherosclerotic process. Presented herein are the results of studying various types of fluctuations, whose absorption bands correspond to such organic compounds as collagen, elastin and cholesterol esters. A peculiar feature of this technique consists in a new method of the preparation of the material, including preparation of samples to be studied, with no stage of fixation. Analysed herein is one of the pathogenetic mechanisms of the development of the atherosclerotic process, consisting in accumulation in the vascular intima of low and very low density lipoproteins which are detected at a frequency of 1745 cm(-1), corresponding to stretching vibrations of the C=O bond. These organic compounds saturate from the inside the vascular wall, thus leading to formation of a lipid nucleus of an atherosclerotic plaque, whose centre is located at a certain depth. The authors present the results of studying the level of the centre of the lipid nucleus of an atherosclerotic plaque in abdominal aorta in perished 70-74-year-old people. It was determined that for this age group the level of the lipid nucleus is concentrated at a depth of 200-240 µm. Besides, presented are the results obtained while studying the abdominal aorta in 23-26-year-old deceased people whose vessels were not affected by atherosclerosis. Obtained are comparative results on the shift in bandwidth of amid I and amid II in studying the intima of the healthy aorta and atherosclerosis-affected aorta. This peculiarity may later on become an identifier of the degree of the development of atherosclerosis in man. The results of such studies provide possibility to influence namely the centre of the lipid nucleus as one of the initial stages of the development of atherosclerosis in order to slow down the progression of the disease concerned.

Host preferences of ornithophilic biting midges of the genus Culicoides in the Eastern Balkans.

Many biting midges of the genus Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) are competent vectors of a diverse number of pathogens. The identification of their feeding behaviour and of vector-host associations is essential for understanding their transmission capacity. By applying two different nested polymerase chain reaction (PCR) assays, of which one targeted the avian cyt b gene and the other targeted the COI gene of a wide range of vertebrates, we identified the blood hosts of six biting midge species including Culicoides circumscriptus, Culicoides festivipennis, Culicoides punctatus, Culicoides pictipennis, Culicoides alazanicus and Culicoides cf. griseidorsum, the latter two of which are reported in Bulgaria for the first time. Bird DNA was found in 50.6% of 95 investigated bloodmeals, whereas mammalian DNA was identified in 13.7%. Two Culicoides species were found to feed on both birds and mammals. There was remarkable diversity in the range of avian hosts: 23 species from four orders were identified in the abdomens of four Culicoides species. The most common bird species identified was the magpie, Pica pica (n = 7), which was registered in all four ornithophilic biting midge species. Six bloodmeals from the great tit, Parus major, were recorded only in C. alazanicus. None of the studied species of Culicoides appeared to be restricted to a single avian host.

Dietary essential oils improve the hepatic antioxidative status of broiler chickens.

1. A total of 200 male Ross 308 chickens were used to evaluate the effects of a standardised combination of essential oils including 5% carvacrol, 3% cinnamaldehyde and 2% capsicum oleoresin (XT 6930; Pancosma S.A., Geneva, Switzerland) on their performance, hepatic antioxidant concentration and caecal tonsils morphometry. 2. Two diets were offered to broiler chickens from d old to 21 d of age. The control diet (C) was slightly lower in metabolisable energy (12.13 MJ/kg ME) and crude protein (215 g/kg CP) than breeders' recommendation. The second diet, made as XT 6930, was added on the top of the control diet at 100 mg/kg. Each diet was offered ad libitum to birds housed in one of 10 floor pens in a randomised complete block design. The birds were housed in 20 floor pens, 10 birds in each pen, and were allocated to 10 replicates of the two dietary treatments. 3. The concentration of antioxidants in the liver of the birds was determined using high-performance liquid chromatography (HPLC) at 21 d of age. Birds fed control diet only had lower weight and converted less efficiently feed to gain compared to birds fed essential oils-supplemented diet. Feed consumption was not affected by dietary treatments. The antioxidant data showed that supplemented essential oils improved the hepatic concentration of carotenoids and coenzyme Q10 when fed to broiler chickens. The morphometry of the caecal tonsils of the birds was not influenced by dietary treatments. 4. It can be concluded that that dietary combination of essential oils, including carvacrol, cinnamaldehyde and capsicum oleoresin, improved growth, feed efficiency and the hepatic concentration of carotenoids and coenzyme Q10 when fed to broiler chickens.

The sensitivity of innovative techniques for measuring low levels of radon in the environment using passive detectors.

In recent years, there has been a significant surge in interest in measuring low radon levels in the environment. These measurements are valuable, particularly for identifying radon priority areas as required by the European Council Directive 2013/59/EURATOM and for research related to climate change. Due to the limited sensitivity of existing radon detectors/monitors in addressing these challenges, substantial efforts have been devoted to developing new designs. This report compares the sensitivity of several innovative designs with that of existing passive radon monitors. These novel designs incorporate alpha track detectors, including large area low background detectors, with activated carbon fabric used as an efficient radon adsorber/radiator. Recent innovative solutions to mitigate the impact of temperature and humidity on detectors using adsorbers are also discussed. The background signal of detectors intended for use in these novel designs is examined, and their sensitivity is evaluated. The findings demonstrate that these novel designs have the potential to significantly enhance the sensitivity of long-term radon measurements, surpassing the detectors currently in widespread use by more than an order of magnitude.

Human immunodeficiency virus type 1 neutralizing antibodies accelerate clearance of cell-free virions from blood plasma.

The concentration of human immunodeficiency virus type 1 (HIV-1) particles in blood plasma is very predictive of the subsequent disease course in an infected individual; its measurement has become one of the most important parameters for monitoring clinical status. Steady-state virus levels in plasma reflect a balance between the rates of virions entering and leaving the peripheral blood. We analyzed the rate of virus clearance in the general circulation in rhesus macaques receiving a continuous infusion of cell-free particles in the presence and absence of virus-specific antibodies. Here we show, by measuring virion RNA, particle-associated p24 Gag protein and virus infectivity, that the clearance of physical and infectious particles from a primary, dual-tropic virus isolate, HIV-1DH12, is very rapid in naive animals, with half-lives ranging from 13 to 26 minutes. In the presence of high-titer HIV-1DH12-specific neutralizing antibodies, the half-life of virion RNA was considerably reduced (to 3.9-7.2 minutes), and infectious virus in the blood became undetectable. Although physical virus particles were eliminated extravascularly, the loss of virus infectivity in the blood reflected the combined effects of extravascular clearance and intravascular inactivation of HIV-1 infectivity due to antibody binding.

Identification of dynamically distinct subpopulations of T lymphocytes that are differentially affected by HIV.

We examined the effects of human immunodeficiency virus infection on the turnover of CD4 and CD8 T lymphocytes in 17 HIV-infected patients by 30 min in vivo pulse labeling with bromodeoxyuridine (BrdU). The percentage of labeled CD4 and CD8 T lymphocytes was initially higher in lymph nodes than in blood. Labeled cells equilibrated between the two compartments within 24 h. Based on mathematical modeling of the dynamics of BrdU-labeled cells in the blood, we identified rapidly and slowly proliferating subpopulations of CD4 and CD8 T lymphocytes. The percentage, but not the decay rate, of labeled CD4 or CD8 cells in the rapidly proliferating pool correlated significantly with plasma HIV RNA levels for both CD4 (r = 0.77, P < 0.001) and CD8 (r = 0.81, P < 0.001) T cells. In six patients there was a geometric mean decrease of greater than 2 logs in HIV levels within 2 to 6 mo after the initiation of highly active antiretroviral therapy; this was associated with a significant decrease in the percentage (but not the decay rate) of labeled cells in the rapidly proliferating pool for both CD4 (P = 0.03) and CD8 (P < 0.001) T lymphocytes. Neither plasma viral levels nor therapy had an effect on the decay rate constants or the percentage of labeled cells in the slowly proliferating pool. Monocyte production was inversely related to viral load (r = -0.56, P = 0.003) and increased with therapy (P = 0.01). These findings demonstrate that HIV does not impair CD4 T cell production but does increase CD4 and CD8 lymphocyte proliferation and death by inducing entry into a rapidly proliferating subpopulation of cells.

Thermal breakage and self-healing of a polymer chain under tensile stress.

We consider the thermal breakage of a tethered polymer chain of discrete segments coupled by Morse potentials under constant tensile stress. The chain dynamics at the onset of fracture is studied analytically by Kramers-Langer multidimensional theory and by extensive molecular dynamics simulations in one dimension (1D) and three dimension (3D) space. Comparison with simulation data in one and three dimensions demonstrates that the Kramers-Langer theory provides good qualitative description of the process of bond scission as caused by a collective unstable mode. We derive distributions of the probability for scission over the successive bonds along the chain which reveal the influence of chain ends on rupture in good agreement with theory. The breakage time distribution of an individual bond is found to follow an exponential law as predicted by theory. Special attention is focused on the recombination (self-healing) of broken bonds. Theoretically derived expressions for the recombination time and distance distributions comply with MD observations and indicate that the energy barrier position crossing is not a good criterion for true rupture. It is shown that the fraction of self-healing bonds increases with rising temperature and friction.

Evidence for cell-surface association between fusin and the CD4-gp120 complex in human cell lines.

Accessory cell-surface molecules involved in the entry of human immunodeficiency virus-type 1 into cells have recently been identified and shown to belong to the family of chemokine receptors. Treatment of human cell lines with soluble monomeric gp120 at 37 degrees C induced an association between the surface CD4-gp120 complex and a 45-kilodalton protein, which can be down-modulated by the phorbol ester phorbol 12-myristate 13-acetate. The three proteins were coprecipitated from the cell membranes with antibodies to CD4 or to gp120. The 45-kilodalton protein comigrated with fusin on sodium dodecyl sulfate gels and reacted with rabbit antisera to fusin in protein immunoblots. No 45-kilodalton protein could be coprecipitated from similarly treated nonhuman cells. However, infection of 3T3.CD4.401 cells with vaccinia-fusin recombinant virus (vCBYF1), followed by gp120 treatment, resulted in coprecipitation of fusin and CD4.401 molecules from their membranes. Together these data provide evidence for physical association between fusin and the CD4-gp120 complex on cell membranes.

Capillary filling in microchannels with wall corrugations: a comparative study of the Concus-Finn criterion by continuum, kinetic, and atomistic approaches.

We study the impact of wall corrugations in microchannels on the process of capillary filling by means of three broadly used methods: computational fluid dynamics (CFD), lattice Boltzmann equations (LBE), and molecular dynamics (MD). The numerical results of these approaches are compared and tested against the Concus-Finn (CF) criterion, which predicts pinning of the contact line at rectangular ridges perpendicular to flow for contact angles of theta > 45 degrees . Whereas for theta = 30, 40 (no flow), and 60 degrees (flow) all methods are found to produce data consistent with the CF criterion, at theta = 50 degrees the numerical experiments provide different results. Whereas the pinning of the liquid front is observed both in the LB and CFD simulations, MD simulations show that molecular fluctuations allow front propagation even above the critical value predicted by the deterministic CF criterion, thereby introducing a sensitivity to the obstacle height.

The escape transition of a polymer: a unique case of non-equivalence between statistical ensembles.

A flexible polymer chain under good solvent conditions, end-grafted on a flat repulsive substrate surface and compressed by a piston of circular cross-section with radius L may undergo the so-called "escape transition" when the height of the piston D above the substrate and the chain length N are in a suitable range. In this transition, the chain conformation changes from a quasi-two-dimensional self-avoiding walk of "blobs" of diameter D to an inhomogeneous "flower" state, consisting of a "stem" (stretched string of blobs extending from the grafting site to the piston border) and a "crown" outside of the confining piston. The theory of this transition is developed using a Landau free-energy approach, based on a suitably defined (global) order parameter and taking also effects due to the finite chain length N into account. The parameters of the theory are determined in terms of known properties of limiting cases (unconfined mushroom, chain confined between infinite parallel walls). Due to the non-existence of a local order parameter density, the transition has very unconventional properties (negative compressibility in equilibrium, non-equivalence between statistical ensembles in the thermodynamic limit, etc.). The reasons for this very unusual behavior are discussed in detail. Using Molecular Dynamics (MD) simulation for a simple bead-spring model, with N in the range 50

Identifying radon priority areas and dwellings with radon exceedances in Bulgaria using stored CD/DVDs.

The implementation of the 2013/59/EURATOM directive in the part related to radon exposure imposes challenges for radon measurement methodology and radon survey design. Among them is the need to have estimates (preferably direct) of the annual average radon concentrations, which can be directly compared to the recommended reference levels. On this basis, the surveys should make possible the identification of dwellings with indoor radon above the reference levels and "radon priority areas" where significant proportion of the dwellings falls in this category. The performance of the CD/DVD method for radon measurements as a tool to address these issues is presented. A recent large scale field study based on the CD/DVD method that was carried out in the suburb area of Sofia, Bulgaria is described. Part of the studied area was affected in the past by the uranium mining and milling industry. In total 462 disks (CDs and DVDs) taken from 335 private dwellings from 10 districts in the region were analyzed. The results revealed the large heterogeneity in radon distribution in the area, with the percentage of dwellings with a 222Rn level above 300 Bq m-3 ranking from about 7% to 74%. The district of Yana, for which this percent was 74, was identified as the area of highest radon priority in the region. The paper also discusses how prompt identification of dwellings with radon above the reference level by CD/DVDs can be incorporated within an integrated approach to the radon problem. Within this approach the radon hazard is identified shortly after the stakeholder's decision to test, which allows fast solution of the problem without waiting the long (and usually demotivating) one-year period needed for direct results by the commonly used prospective methods.

Optimization and diagnostic evaluation of monoclonal antibody-based blocking ELISA formats for detection of neutralizing antibodies to Hendra virus in mammalian sera.

Maintenance of Hendra virus (HeV) in pteropid bat populations has been associated with spillover events in horses, humans and dogs. Experimental studies have demonstrated infections for several other species including guinea pigs, cats and ferrets. The criteria of a sensitive and specific serological test that is effective for a range of species, but which does not require use of live virus, has not been satisfactorily addressed by currently available tests. We have evaluated the use of two HeV neutralizing monoclonal antibodies (mAbs) in a blocking format enzyme-linked immunosorbent assay (bELISA) to detect serum antibody against a recombinant expressed HeV G protein (sol G) in several animal species. The human mAb m102.4 neutralises both HeV and the closely related Nipah virus (NiV); the mouse mAb 1.2 neutralises only HeV. Given these functional differences, we have investigated both antibodies using a bELISA format. Diagnostic sensitivity (DSe) and diagnostic specificity (DSp) were optimized using individual thresholds for mAb 1.2 and m102.4. For mAb 1.2 the positive threshold of >33% inhibition yielded DSe and DSp values of 100% (95% CI 95.3-100.0) and 99.5 (95% CI 98.8-99.8) respectively; for mAb m102.4 a positive threshold of >49% inhibition gave DSe and DSp values of 100 (95% CI 95.3-100.0) and 99.8 (95% CI 99.2-100.0) respectively. At these thresholds the DSe was 100% for both tests relative to the virus neutralization test. Importantly, the occurrence of false positive reactions did not overlap across the assays. Therefore, by sequential and selective application of these assays, it is possible to identify false positive reactions and achieve a DSp that approximates 100% in the test population.

Universal properties of a single polymer chain in slit: Scaling versus molecular dynamics simulations.

We revisit the classical problem of a polymer confined in a slit in both of its static and dynamic aspects. We confirm a number of well known scaling predictions and analyze their range of validity by means of comprehensive molecular dynamics simulations using a coarse-grained bead-spring model of a flexible polymer chain. The normal and parallel components of the average end-to-end distance, mean radius of gyration and their distributions, the density profile, the force exerted on the slit walls, and the local bond orientation characteristics are obtained in slits of width D=4/10 (in units of the bead diameter) and for chain lengths N=50/300. We demonstrate that a wide range of static chain properties in normal direction can be described quantitatively by analytic model-independent expressions in perfect agreement with computer experiment. In particular, the observed profile of confinement-induced bond orientation is shown to closely match theory predictions. The anisotropy of confinement is found to be manifested most dramatically in the dynamic behavior of the polymer chain. We examine the relation between characteristic times for translational diffusion and lateral relaxation. It is demonstrated that the scaling predictions for lateral and normal relaxation times are in good agreement with our observations. A novel feature is the observed coupling of normal and lateral modes with two vastly different relaxation times. We show that the impact of grafting on lateral relaxation is equivalent to doubling the chain length.

Dynamics of a stretched nonlinear polymer chain.

We study the relaxation dynamics of a coarse-grained polymer chain at different degrees of stretching by both analytical means and numerical simulations. The macromolecule is modeled as a string of beads, connected by anharmonic springs, subject to a tensile force applied at the end monomer of the chain while the other end is fixed at the origin of coordinates. The impact of bond nonlinearity on the relaxation dynamics of the polymer at different degrees of stretching is treated analytically within the Gaussian self-consistent (GSC) approach and then compared to simulation results derived from two different methods: Monte Carlo (MC) and Molecular Dynamics (MD). At low and medium degrees of chain elongation we find good agreement between GSC predictions and the MC simulations. However, for strongly stretched chains, the MD method, which takes into account inertial effects, reveals two important aspects of the nonlinear interaction between monomers: (i) a coupling and energy transfer between the damped, oscillatory normal modes of the chain and (ii) the appearance of nonvanishing contributions of a continuum of frequencies around the characteristic modes in the power spectrum of the normal mode correlation functions.

Subdermal laterocranial mapping of sentinel lymph nodes in breast cancer as an alternative to the peritumoral mapping.

PURPOSE: Peritumoral injection of blue dye is standard method of sentinel lymph nodes (SLN) biopsy in breast cancer. Applying the marker to different locations will be of great benefit in cases in which the peritumoral mapping is difficult to implement. The aim of this study was to demonstrate that the subdermal laterocranial (SLC) mapping with a dye has similar diagnostic reliability to the peritumoral mapping and is applicable for clinical use. PATIENTS AND METHODS: 254 patients with operable breast cancer (cT

THE CD/DVD METHOD AS A TOOL FOR THE HEALTH PHYSICS SERVICE AND VENTILATION DIAGNOSTICS IN UNDERGROUND MINES.

The CD/DVDs used as radon (222Rn) and thoron (220Rn) detectors can provide a sufficiently sensitive and cost-efficient option for passive radiation monitoring in underground mines. This note presents results of measurements made under real environmental conditions by CD/DVDs. Comparison with conventional diffusion chambers was made and good correspondence was observed. Correlation between 222Rn and 220Rn was studied by CD/DVDs and no signs for any correlation were observed. Dedicated study in a mine gallery showed that CD/DVDs can be successfully used for the purposes of ventilation diagnostics by identifying sources of air contaminated with radon.

Capillary rise in nanopores: molecular dynamics evidence for the Lucas-Washburn equation.

When a capillary is inserted into a liquid, the liquid will rapidly flow into it. This phenomenon, well studied and understood on the macroscale, is investigated by molecular dynamics simulations for coarse-grained models of nanotubes. Both a simple Lennard-Jones fluid and a model for a polymer melt are considered. In both cases after a transient period (of a few nanoseconds) the meniscus rises according to a (time)1/2 law. For the polymer melt, however, we find that the capillary flow exhibits a slip length delta, comparable in size with the nanotube radius R. We show that a consistent description of the imbibition process in nanotubes is only possible upon modification of the Lucas-Washburn law which takes explicitly into account the slip length delta. We also demonstrate that the velocity field of the rising fluid close to the interface is not a simple diffusive spreading.

COMPARATIVE STUDY OF RADON AND THORON MEASUREMENTS IN FOUR ROMANIAN SHOW CAVES.

Measurements have been carried out using four types of passive detectors in four of the most popular show caves in Romania. Three types of detectors (RSKS, RadTrak and CD) were used for radon measurements and two (Raduet and CD) for thoron measurement. Activity concentrations in air were measured in the same locations for two seasons, autumn and winter. Measured values for the different caves varied between below detection limit (5 Bq m-3) and 4024 Bq m-3 for radon and from below 10 to 583 Bq m-3 for thoron. The results indicate a very good correlation between RSKS and RadTrak detectors (r = 0.96). The most significant difference between radon concentrations measured with different types of detectors (RSKS and CD) was higher than 150%. The study suggests that the activity concentration of radon in caves, measured using track detectors, could not be influenced by the type of detector used if the microclimate factor is acknowledged.