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The aim of this work is to investigate the dynamics for the ovarian tissue engraftment of inbred August rats transplanted to outbred Wistar rats and vice versa on the background of the induction of donor-specific tolerance and to identify the features of proteasome pools in the surviving grafts and tissue replacing the rejected grafts in animals of both groups. By day 14 after transplantation, a slight difference in graft engraftment was revealed in studied recipients: 87% in Wistar rats and 80% in August rats. At the same time, ovarian tissue allografts with well-preserved structure and significant vascularization represented 76% in Wistar rats and only 29% in August rats on day 37. The difference revealed in the remote period is apparently connected with the special condition of the central nervous system of August rats caused by the increased content of monoamines and heat-shock protein 70. The total proteasome level on day 37 was the same both in intact donor tissue and in the surviving grafts, as well as in the tissue replacing the rejected grafts, and did not depend on the donor–recipient differences. However, the increased expression of immune proteasomes was found in the surviving tissue and tissue replacing the rejected grafts compared to intact donor tissue. The surviving grafts were enriched with proteasomes with immune subunit LMP2, and the tissue replacing the rejected grafts was enriched with proteasomes with immune subunit LMP7 equally in August and Wistar rats. In addition, the surviving grafts in both groups of animals were characterized by equally high expression of proteasome activator РА28αβ compared to the replacement and intact donor tissues. Thus, transplant proteasomes containing immune subunits LMP2, and probably associated with the activator RA28αβ, serve as one of the key factors for engraftment regardless of the genetically determined differences of the recipients. [ABSTRACT FROM AUTHOR]

The aim of the work was to standardize the conditions for obtaining viable and functionally complete cells in the composition of multicellular spheroids (MS). The latter were obtained in the conditions of 7 days 3D cultivation with fibroblasts seeding at concentrations of 1·105, 2·105 and 5·105 cells/ ml. It has been proved that the sowing concentration of fibroblasts affects the size of the formed MS as well as the viability, morphological parameters, proliferative and adhesive properties of cells they are composed of. The maximal concentration that have been used had a negative impact on spheroid formation, on the ability of cells to adhesion. This concentration reduced cell viability by 19%, coefficient of increasing the number of cells 4,25-fold and increased the number of cells with nuclear fragmentation and chromatin condensation by 20% in comparison with the sowing concentration of 2·105 cells/ml. At a concentration of 1·105 cells/ml, predominantly single and small (about 10-20 mkm) MS with a cell viability of about 89,5 ± 1,2% were formed. The fibroblasts had normal morphological characteristics, but the coefficient of increasing the number of cells was significantly lower than the similar indicator at the sowing concentration of 2·105 cells/ml. Thus, it has been established that the concentration of 2·105 cells/ml is optimal for the formation of MS with morphologically and functionally complete fibroblasts. [ABSTRACT FROM AUTHOR]

Induction of donor-specific tolerance in outbred Wistar rats (RT1u) and inbred August rats (RT1c) increased the expression of immune proteasome subunits in liver with a peak on day 7 after beginning of the induction. The increase in the level of immune subunits LMP2 and LMP7 was more pronounced in the liver of August rats in comparison with Wistar rats (by 2 and 6 times, respectively), which was associated with higher concentrations of monoamines in the CNS of August rats. After induction of donor-specific tolerance in August and Wistar rats, the immune subunits were in cells of sinusoidal lining and in cells located in sinusoid lumens. It can be suggested that immune proteasomes in these cells producing antigenic peptides for presentation to immunocompetent participate in the suppression of their activity and form the molecular basis for the development of donor-specific tolerance at very early stages of this process. [ABSTRACT FROM AUTHOR]

Basic research of the structure and functions of multiple forms of proteasomes is overviewed. The proteasomes of liver, acting as a primary lymphoid organ in embryogenesis, displaying nonspecific local immunity, and providing development of tolerance to food and other foreign antigens during the lifespan, are described. The role of individual forms of liver proteasomes in transplantology is considered as well as their utility as markers for endocrine transplant survival. [ABSTRACT FROM AUTHOR]

Background: The Ukrainian American Cohort Study was established to evaluate the risk of thyroid disorders in a group exposed as children and adolescents to 131I by the Chernobyl accident (arithmetic mean thyroid dose, 0.79 grays). Individuals are screened by palpation and ultrasound and are referred to surgery according to fine-needle aspiration biopsy (FNA). However, the accuracy of FNA cytology for detecting histopathologically confirmed malignancy after this level of internal exposure to radioiodines is unknown.Methods: During the first screening cycle (1998-2000), 13,243 individuals were examined, 356 individuals with thyroid nodules were referred for FNA, 288 individuals completed the procedure, 85 individuals were referred to surgery, 82 individuals underwent surgery, and preoperative cytology was available for review in 78 individuals. Cytologic interpretation for the nodule that resulted in surgical referral was correlated with final pathomorphology; discrepancies were reviewed retrospectively; and the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of FNA cytology were calculated.Results: All 24 cytologic interpretations that were definite for papillary thyroid cancer (PTC) were confirmed histopathologically (PPV, 100%); and, of 11 cytologic interpretations that were suspicious for PTC, 10 were confirmed (PPV, 90.9%). Ten of 41 FNAs that were interpreted as either definite or suspect for follicular neoplasm were confirmed as malignant (PPV, 24.4%), including 2 follicular thyroid cancers and 8 PTCs (all but 1 of the follicular or mixed subtypes). Depending on whether a cytologic interpretation of follicular neoplasm was considered "positive" or "negative," the sensitivity was 100% and 77.3%, respectively; similarly, the respective specificity was 17.6% and 97.1%, the respective PPV was 61.1% and 97.1%, and the respective NPV was 100% and 76.7%.Conclusions: Among children and adolescents who were exposed to 131I after the Chernobyl accident and were evaluated 12 to 14 years later, thyroid cytology had a sensitivity and a predictive value similar to those reported in unexposed populations. [ABSTRACT FROM AUTHOR]

Nine novel 6-fluorothymine nucleoside analogues of both N(1)-α/β and N(3)-β-ribo series were prepared by the Vorbrüggen method starting from persilylated 6-fluorothymine and 1-O-acetyl-2,3,5-tri-O-benzoyl-β- d -ribofuranose, 1-O-acetyl-2,5-di-O-benzoyl-3-deoxy-3-fluoro-α,β- d -ribofuranose or 1,2,3,5-tetra-O-benzoyl-β- d -ribofuranose and its α-anomer. Protected N(3)-β- d -ribofuranosides were prepared as sole products in high yields at room temperature. A mixture of benzoylated N(1)-β- and α-anomeric ribonucleosides was obtained at lower temperatures. Yields of β-anomers and stereoselectivities (β:α=2.2/4.5:1) of the condensation reactions depended on reaction conditions and the structure of the glycosylating agent. Debenzoylation of 6-fluorothymine N(1)- or N(3)-β- d -ribofuranosides and their 3′-fluorodeoxy analogues by LiOH monohydrate in MeCN/H 2 O resulted in the corresponding fluorinated nucleosides in good yields, whereas the deprotection of N(1)-α-ribofuranosides under the same conditions unexpectedly yielded 6,2′-O-α- d -anhydronucleosides. 6-Substituted (OMe, NH 2 ) thymine β-ribonucleosides were prepared by the treatment of protected N(1)-β- d -ribosides with nucleophilic agents. [ABSTRACT FROM AUTHOR]

Stem/progenitor cells are thought to have the potential in the treatment of severe neurodegenerative diseases. Recently, sympathoadrenal progenitors expressing specific markers of neural crest derivatives and capable to differentiate into neurons were discovered in adult bovine and human adrenal glands, but there was no reported data on cryopreservation of sympathoadrenal progenitors. The aim of the present study was to examine the neural differentiation potential of sympathoadrenal progenitors derived from fresh and cryopreserved neonatal porcine adrenal glands. Considering impact of various initial state of frozen biomaterial on cell recovery, we carried out a comparative estimation of cryopreservation outcome both for adrenal tissue fragments and isolated primary cells. The estimation consisted of determining cell yield, viability, ability to adhere, proliferate and differentiate in vitro. Cells isolated from the fresh adrenal glands were cultured until confluence. A formation of sympathoadrenal progenitors-embedded spherical cell colonies, whose cells are differentiated then into βIII-tubulin-positive cells with neuron-like morphology, was observed on the monolayer. The colonies were well preserved after cryopreservation of cell culture with a cooling rate of 1 °C/min in the cryoprotectant media containing 5–15% of dimethylsulfoxide. Adrenal tissue fragments were cryopreserved in the presence of 10% dimethylsulfoxide at the cooling rates of 0.3; 1: 5; 40 and > 100 °C/min. Sympathoadrenal progenitors were recovered after cryopreservation with 0.3 °C/min cooling rate but not higher. [ABSTRACT FROM AUTHOR]

Purpose: Forwarding service is a key part of the system of goods movement and as such it provides realization of numerous services. Providing of new additional and unconventional services to cargo owners by haulers will extend the range of their clients, increase profits from sales, facilitate and accelerate the introduction of new transport technologies, increase stability and strength of relations in the transport market. Methods: We formulated requirements to be met by modern forwarding service. Carriage management should be based on logistics concept and its methods; comprehensive resource optimization of haulers and owners of cargo and other supply chain participants (insurance, financial ones, etc.). Optimization of the space-time chain, even involving freight forwarders (transport intermediaries), is a complex scientific and practical task. Results: We offer directions for development of forwarding additional and non-conventional service for forwarding system of cargo haulage service, which would also provide an opportunity to reduce transport time and increase service level. Discussion: A general scheme of products delivery to consumers contains a significant number of units-particular entities. It is important to analyze not only the position of economic entities in the market, but the features and capabilities of their interaction in major systemic tasks of forwarding service, quality and effective promotion of freight flows. [ABSTRACT FROM AUTHOR]

New azacytidine nucleoside analogs with modified carbohydrate moieties were synthesized. Screening identified a highly active 2′-fluoro-containing azacytidine analog that could potentially be of interest as an agent for treating acute myelogenous leukemia and myelodysplastic syndrome. [ABSTRACT FROM AUTHOR]

Abstract: Cryopreservation is thought to have the potential to preserve tissue for transplantation. In addition, it can also be used for decreasing tissue immunogenicity, which might be important for prolonging allograft survival. In the present study we examined the impact of cryopreservation at various cooling rates on the outcome of allotransplantation of murine adrenal tissue fragments (ATFr). ATFr were cryopreserved with a cooling rate at 1; 10; 40 and more than 100°C/min. After thawing it was found that the number of the cells expressing markers of dendritic cells (CD11c) and macrophages (CD11b) in the suspension obtained from ATFr decreased with increasing cooling rate. After allotransplantation the survival rates of adrenalectomized mice and the blood serum levels of corticosterone were higher in recipients of cryopreserved ATFr. By immunohistochemistry, cryopreserved allografts displayed a decreased infiltration by CD4+ and CD8+ T-lymphocytes as compared to fresh grafts. These findings suggest that cryopreserved allografts cause a less severe rejection by decreasing graft immunogenicity. [Copyright &y& Elsevier]

Induction of donor-specific tolerance in a recipient is one of the methods for enhancing acceptance of the grafts of endocrine glands in the absence of immunodepressants, which interfere with hormone production. This paper describes changes in the proteasome pool in the rat liver, spleen, and graft during the development of donor-specific tolerance after intraportally infusing the recipient with donor splenocytes with subsequent allografting of ovarian tissue into the renal capsule. It has been demonstrated that the shift in the balance in the liver and graft proteasome pools towards the variants with the LMP2 subunit determines the development of immunological tolerance and graft retention. On the contrary, an increase in the forms with the LMP7 subunit induces the immune response and graft rejection. [ABSTRACT FROM AUTHOR]

We obtain a formula for the determination of a defect under a continuous imbedding of subspaces in the scale of Hilbert spaces. [ABSTRACT FROM AUTHOR]

We discuss a new approach in singular perturbation theory which is based on the method of rigged Hilbert spaces. Let A be a self-adjoint unbounded operator in a state space and ▪ be the rigged Hilbert space associated with A in the sense that in the graph-norm. We propose to define the perturbed operator à as the self-adjoint operator uniquely associated with a new rigged Hilbert space ▪ constructed using a given perturbation of A. We show that the well-known form-sum and self-adjoint extensions methods are included in the above construction. Moreover, we show that the super singular perturbations may also be described in our framework. [Copyright &y& Elsevier]

Let A be an unbounded self-adjoint operator in a Hilbert separable space $$H_0$$ with rigging $$H_ - \sqsupset H_0 \sqsupset H_ +$$ such that $$D(A) = H_ +$$ in the graph norm (here, $$D(A)$$ is the domain of definition of A). Assume that $$H_ +$$ is decomposed into the orthogonal sum $$H_ + = M \oplus N_ +$$ so that the subspace $$M_ +$$ is dense in $$H_0$$ . We construct and study a singularly perturbed operator A associated with a new rigging $$H_ - \sqsupset H_0 \sqsupset \overset{\lower0.5em\hbox{$\smash{\scriptscriptstyle\smile}$}}{H} _ +$$ , where $$\overset{\lower0.5em\hbox{$\smash{\scriptscriptstyle\smile}$}}{H} _ + = M_ + = D(\overset{\lower0.5em\hbox{$\smash{\scriptscriptstyle\smile}$}}{A} )$$ , and establish the relationship between the operators A and A. [ABSTRACT FROM AUTHOR]

Abstract Carp, Cyprinus carpio L., spermatozoa swelling in NaCl hypotonic solutions (18.8, 37.5, 75, 100 mM) was studied by the methods of photometry and resistance impulses spectroscopy (RIS). The possibility of application of the value of relative optical density of cell suspension, as a qualitative characteristics of the extent of spermatozoa swelling in the period of their movement, was demonstrated. It has been noted, that the moment when swelling starts coincides with the moment of the start of sperm motility. Movement duration, extent and time of swelling depend on the osmotic pressure of the activation solution and moreover the maximum motile period is observed during the activation by 75 mM NaCl solution. [ABSTRACT FROM AUTHOR]

• Symmetry is an important aspect of nuclear shape not receiving the attention it merits. • A software, nPAsym, implemented as ImageJ plugin is presented allowing to measure nuclear shape asymmetry in digital images captured from cytologic and histologic preparations. • nPAsym has proved itself effective in distinguishing between some of the nuclear shape phenotypes. • nPAsym has a potential to become useful for both experimental research and diagnostics. : The nucleus is a complex and dynamic organelle enclosing the major part of the cell's genome. A growing body of evidence suggests that changes in the shape of this organelle can influence cell activities. The other way around, altered nuclear shape may be indicative of impaired cell function. Symmetry is an important aspect of nuclear shape not receiving the attention it merits. We address this problem by presenting a software tool allowing to quantify nuclear shape asymmetry in light microscopy images. : The software named nPAsym is written in Scala and implemented as a plugin to ImageJ making possible to use it in combination with other ImageJ tools. The plugin works with 8-bit images segmented into black nuclear masks and white background. It performs a number of operations allowing to analyze multiple objects within a single image, removing some segmentation artefacts, filtering out objects incomplete and below a specified size. The feature of interest is quantified using the notion of point asymmetry. The performance of nPAsym was tested in a small-scale study comparing nuclear shapes for cells of nodular goiter, follicular thyroid adenoma and papillary thyroid carcinoma. : We present nPAsym, the ImageJ plugin, that measures nuclear shape asymmetry. It works with digital microscopic images segmented using either a raster graphics editor or built-in ImageJ functions. nPAsym is packaged in a single.jar file and does not require installation as well as configuration. It has proved effective in distinguishing between some of the nuclear shape phenotypes. : nPAsym is the user-friendly, platform-independent and open-source software tool allowing to quantify nuclear shape asymmetry in digital images captured from cytologic and histologic preparations. It has a potential to become useful for both experimental research and diagnostics. [ABSTRACT FROM AUTHOR]

Lensless inline digital holographic microscopy (LI-DHM) and Fourier ptychographic microscopy (FPM) are two widespread quantitative phase imaging (QPI) techniques. They have been employed in various fields, especially for biological slice imaging because of their simplicity in use, stability in structure, and also large field of view. Spherical phase response (for example from HeLa cells) is commonly observed in biological imagery. As a consequence, for calibration and validation purposes, small (several to tenth of microns in diameter) transparent microbeads have been used as standards. Phase imaging of their large counterparts (hundreds of microns in diameter) using either LI-DHM or FPM has not been reported so far. We are aiming to analyze the phase response of a 146-μm soda-lime microsphere. It has been immersed in Canada balsam to reduce phase difference and to avoid overexposed diffraction rings. The phase estimation issue has been tackled using approaches that involve either Gerchberg–Saxton type algorithms or an inverse problem-based procedure. Confronting the results confirms the QPI capability for both imaging techniques to assess phase responses from such a large transparent object. [ABSTRACT FROM AUTHOR]

Background. The dermal papilla (DP) formation is associated with a complex process of neural crest cells migration. DP contains pluripotent cells that support renewal of a hair follicle over a long period. The possibility of culturing DP cells of several species (mouse, rat, human, and sheep) was shown. The aim of our study was to demonstrate the possibility of culturing DP cells of rabbit vibrissae. Methods. Cell cultures were isolated from explants by the method of Sieber-Blum, 2004. The isolated DP were placed in 6-well culture plates (PAA, Austria) and passaged every 7 days. It was investigated the possibility of obtaining different types of culture, such as monolayer and 3D culture of multicellular spheroids (MSs). The monolayer was cultured in the medium DMEM/F12 with 10 % PBS (Biovest, France). This type of culture has a high proliferative potential; the proliferation rate on day 3 is 9-12. However, it is believed that the 3D cell culture of multicellular spheroids is the form in which cells better retain their uncommitted state (Santana et al., 2012). To obtain MSs, we used Petri dishes with low-adhesion surface (PAA, Austria). The DMEM/F12 culture medium with addition of 2 % [of] supplements (B27 or Neuromax (Pan Eco, RF) was used in this case. Results. The formation of MS was observed when using both growth supplements. The formation of spheroids occurred by aggregation of single floating cells. The clusters formation started in several hours after passaging, a day later they contained 10–15 cells and acquired a spherical shape. During culture, the MSs were compacted and reached [the] sizes of 100-150 nm in 14 days. It was found that the cells in MS preserved the ability to grow in a monolayer. MS was transferred to [the] culture medium supplemented with 10 % FBS on days 10-14 of cultivation, after which the adhesion cells started to migrate onto the substrate. Conclusions. Thus, the possibility of culturing DP cells of the vibrissa rabbit in vitro was shown. It is possible to obtain both monolayer and MS culture using different growth factors. The possibility to use Neuromax for this type of culture as an alternative to the frequently used nutritional supplement B27 was also shown. [ABSTRACT FROM AUTHOR]

The development of inexpensive equipment adapted for the study of a specific biological object is very important for cryobiology. In the presented work, we have proposed a simple system for microscopy utilising open‐source platform Arduino. Testing this system showed that it had sufficient sensitivity to determine the physical processes occurring in a cryopreserved sample such as intra‐ and extracellular water crystallisation and salt eutectic. Utilising this system, we investigated the mechanisms of cryoprotection and cryodamage of testis interstitial cells (ICs) in cryoprotective media, which included cryoprotective agents such as dimethyl sulphoxide (Me2SO), as well as foetal bovine serum or polymers (dextran, hydroxyethyl starch and polyethylene glycol). It was shown that a serum‐/xeno‐free medium that included 0.7 M Me2SO and 100 mg/mL dextran was able to reduce intracellular water crystallisation in cells, change the structure of extracellular ice, and reduce salt eutectic and recrystallisation. All these effects correlated with better IC survival after cryopreservation in the medium. This medium is potentially less toxic as it has lower concentrations of Me2SO compared to serum‐containing media developed for cryopreservation of testicular cells. This would pave a way for the creation of nontoxic serum‐free compositions that does not require removal before use of cryopreserved living cells for laboratory practice or in clinics. LAY DESCRIPTION: Research in the field of cryobiology involves many technics linked with the assessment of cell recovery after cooling/warming as well as quantification, identification and description of physical processes occurring in and out of the cells. These processes can affect the overall outcome of cryopreservation. Thus, the investigation of the mechanisms of cryoprotection and cryodamage is of high importance. In this work, we have developed a system for cryomicroscopy utilising open‐source platform for prototyping electronic devices and systems Arduino. The platform includes hardware and software designed to develop various projects. It also supports a programming language. The language was used to create a code to control cooling rate of the sample undergoing microscopy. The system was able to visualise the structure of the forming ice crystals, recrystallisation, salt eutectic crystallisation/melting and intracellular phase transitions. It can be utilised in combination with other approaches and techniques. The system helped us to solve specific tasks of cryobiological experimentation connected with the establishment of mechanisms of cryoprotection and cryodamage of testis interstitial cells (ICs). The microscopy data allowed us to compare different media developed for cryopreservation of testicular cells. The serum‐containing cryoprotective medium (1.4 M Me2SO and 10% foetal bowine serum) was able to suppress darkening of cell, known as 'flashing' associated with intracellular phase transitions by moderately dehydrating cells, and salt eutectic transitions during cooling/warming. However, the media contains relatively high Me2SO concentration and may be toxic. Additionally, the use of serum in the media can result in transferring infection when cryopreserved cells are used in medicine or veterinary, or lead to instability of the composition of serum‐containing media from batch to batch. In the course of the research, we have shown that using the serum‐/xeno‐free medium supplemented with 0.7 M Me2SO and 100 mg/mL Dex (0.7 Me2SO+Dex) can also reduce cell 'flashing'. The medium influenced the size and shape of extracellular ice crystals in a way that can potentially reduce the risk of mechanical cell damage. This medium promoted amorphous solidification in the space between the extracellular ice crystals, thus, lowering the salt eutectic transitions and recrystallisation that could otherwise cause additional cell damage. Therefore, application of the developed system for cryomicroscopy, which allowed real‐time visualisation of cryobiological processes, made it possible to draw a conclusion about cryoprotecitive properties of medium for ICs. 0.7 Me2SO+Dex can be the basis for the creation of commercial serum‐free media with a defined composition that does not require removal before use. [ABSTRACT FROM AUTHOR]

Objective. To investigate the mechanisms of cryoprotection with compositions that were supplemented with polymers (dextran [Dex], hydroxyethyl starch [HES], polyethylene glycol [PEG]) or fetal bovine serum [FBS]). Methods. Differential scanning calorimetry (DSC) and thermomechanical analysis (TMA) of cryoprotective media containing serum, as well as media that do not contain serum or its components but include polymers. Results. The combinations of dimethyl sulfoxide with FBS or polymers in cryoprotective media were able to promote the formation of amorphous phase and suppress salt eutectic crystallization/melting and recrystallization. The medium supplemented with 0.7 M Me2SO and 100 mg/ml Dex (0.7Me2SO+Dex) was somewhat superior compared to other compositions. It also had an optimal glass transition temperature (-26...-21 °C). The feature correlated with an adequate cell recovery when the medium was utilized to preserve testicular cells. Conclusions. The mechanism of cryoprotection depends not only on the concentrations of cryoprotective agents such as Me2SO, polymers or FBS, but also on the composition of the cryoprotective media. The 0.7Me2SO+Dex medium can be the basis for the creation of commercial serum-free cryoprotective media with a defined stable composition that does not require removal of Me2SO before using the cryopreserved cells or tissues for cultivation, transplantation and transfusion. [ABSTRACT FROM AUTHOR]

An abstract of the article "A Cohort Study of Thyroid Cancer and Other Thyroid Diseases After the Chernobyl Accident. Cyto-Histopathologic Correlation and Accuracy of Fine Needle Aspiration Biopsy in Nodules Detected During the First Screening Cycle in Ukraine (1998-2000)," by Ellen Greenebaum, Yuriy M. Bozhok, Tetyana I. Bogdanova, Anna Zelinskaya, Lyudmyla Zurnadzhy, Victor Shpak, Ovsiy V. Epstein, Robert J. Mcconnell, Alina Brenner, Jacob Robbins, Lydia Zablotska, Mykola D. Tronko and Maureen Hatch is presented.

The development of serum-/xeno-free media may help avoid the drawbacks of using serum and its components, such as probable contamination, instability of composition, or difficulty in sterilization. The objectives of this research were to investigate the use of combinations of a permeating cryoprotective agent (Me 2 SO) and non-permeating polymers (polyvinyl alcohol, polyvinylpyrrolidone, polyethylene glycol, hydroxyethyl starch, dextran) for cryopreservation of interstitial cells (ICs) of rat testis, and to propose the mechanism of cryoprotection of such compositions. In the course of this study, the best combination was 100 mg/ml dextran (M.m. 40 kDa) (Dex40) with 0.7 M Me 2 SO in Ham's F12. The ICs were additionally cooled and warmed to different end temperatures (−30, −50, −50 and −196 °C) to determine which temperature intervals contributed most to the IC loss. Then, the cryoprotective action of this serum-/xeno-free medium was investigated in comparison with serum or albumin-containing media by differential scanning calorimetry (DSC) and thermomechanical analysis (TMA). The results showed that the medium based on Dex40 did not decrease the amount of ice formed. However, it could undergo other phase separation and phase transformation to form glassy states. Potential cell-damaging physical processes such as eutectic crystallization/melting, recrystallization of NaCl and/or Me 2 SO derivatives, found in serum-containing media and taking place in specific temperature intervals, were not observed in the Dex40 based media. This was in good correlation with indicators of cell survival. Additionally, the application of Dex40 allowed using Me 2 SO in lower concentrations (0.7 M) than required for serum-containing media (1.4 M), which may decrease the toxicity of serum-/xeno-free media. • This study examined the effect of combinations of Me 2 SO and polymers on testis interstitial cell cryopreservation. • The best combination was 100 mg/ml dextran 40 with 0.7 M Me 2 SO in Ham's F12. • Physical processes in the medium were investigated by differential scanning calorimetry and thermomechanical analysis. • The mechanisms of cryoprotection of this serum-/xeno-free medium based on dextran 40 were proposed. [ABSTRACT FROM AUTHOR]

Proteasomes in the liver of August rats (RT1) were investigated 30 days after allotransplantation of Wistar rat (RT1) thyroid tissue under renal capsule with/without induction of donor-specific tolerance by donor splenocyte intraportal administration. The levels of total proteasome pool, immune proteasomes containing subunits LMP2 and/or LMP7, and proteasome regulators 19S and 11S were defined. Intact and sham-operated August rats were used as control groups. The level of all immune proteasome forms and 11S regulator increased while the level of the total proteasome pool and 19S regulator decreased in the liver of experimental animals compared to the control groups, which indicated changes of liver functional state after transplantation. The 19S/11S ratio increased in the liver of nontolerant rats compared to tolerant animals. In the liver of tolerant rats with accepted grafts, the number of mononuclear cells expressing the immune subunit LMP2 greatly increased in comparison with control and nontolerant animals. Study of accepted grafts showed an increase in the ratio of LMP2/LMP7 immune subunits and 19S/11S regulators in them, compared to the tissue replacing the rejected grafts. Immune proteasomes were almost completely absent from the control intact thyroid tissue, while 19S/11S ratio was maximal in it. Thus, the development of the immune reaction or its suppression are accompanied by a change in the balance between different proteasome forms. Immune subunit LMP7 and 11S regulator are associated with the response against donor tissue. On the contrary, immune subunit LMP2 and 19S regulator are likely to be important for the development of immune tolerance and surviving tissue functioning. Immunofluorescence assay revealed a low content of the immune proteasomes in the follicle cells. Probably, formation of antigens for the major histocompatibility complex class I molecules was impaired by the low content of immune proteasomes, which led to immunological tolerance of hormone-producing follicle cells. [ABSTRACT FROM AUTHOR]

A novel class of potential protein kinase inhibitors 7–16 was synthesized in high yields using various substituted purines. The most promising compounds, 7 and 10, exhibited inhibitory activity against seven cancer cell lines. The IC50 values for compounds 7 and 10 were 2.27 and 2.53 μM for K562 cells, 1.42 and 1.52 μM for HL-60 cells, and 4.56 and 24.77 μM for OKP-GS cells, respectively. In addition, compounds 7 and 10 dose-dependently induced the apoptosis and cell cycle arrest at G2/M phase, preventing the cell division of OKP-GS cells. Compounds 7, 9, and 10 showed 36–45% inhibitory activity against PDGFRα and PDGFRβ at the concentration of 1 μM. Molecular modeling experiments showed that obtained compounds could bind to PDGFRα as either type 1 (compound 7, ATP-competitive) or type 2 (compound 10, allosteric) inhibitors, depending on the substituent in the amide part of the molecule. [ABSTRACT FROM AUTHOR]

Treatment of ethyl 7-amino-3- tert-butyl-4-oxo-4,6-dihydropyrazolo[5,1- c][1,2,4]triazine-8-carboxylate with PS in pyridine gave ethyl 7-amino-3- tert-butyl-4-thioxo-4,6-dihydropyrazolo[5,1- c][1,2,4]triazine-8-carboxylate which was subjected to acylation, decarboxylation, and hydrazinolysis. [ABSTRACT FROM AUTHOR]

Implantation of ovarian tissue allografts in outbred Wistar rats and inbred August rats against the background of induction of donor-specific tolerance was accompanied by an increase in liver content of mononuclear cells expressing LMP2 immune of proteasome subunit by day 37 after transplantation in comparison with day 0. Graft rejection, on the contrary, was associated with a decrease in the number of LMP2+ cells in the liver of rats of both lines during this period. The difference in the content of these cells and the graft take rate were higher in Wistar rats. The number of mononuclear cells expressing LMP7 immune proteasome subunit in the liver did not change in rats of both lines by day 37 in comparison with day 0. Thus, the level of immune proteasomes with LMP2 subunit in mononuclear cells of the liver is related to fine mechanisms of regulation of immune responses and their shift toward graft take or rejection. [ABSTRACT FROM AUTHOR]

Предложена классификация узкоколейных трансферов и туристических путешествий в зависимости от цели поездки, вида подвижного состава, времени путешествия и регионов обслуживания, которая позволит заранее планировать необходимое количество подвижного состава. Определены принципы и ключевые факторы, которые влияют на развитие туристических перевозок по узкоколейным железным дорогам. Рассчитаны прогнозные показатели туристического потока в Закарпатской области и ожидаемый годовой доход от туристических перевозок, которые будут организованы по узкоколейной железной дороге. [ABSTRACT FROM AUTHOR]

Summary: Background: In our randomised, controlled, phase 3 trial NeOAdjuvant Herceptin (NOAH) trial in women with HER2-positive locally advanced or inflammatory breast cancer, neoadjuvant trastuzumab significantly improved pathological complete response rate and event-free survival. We report updated results from our primary analysis to establish the long-term benefit of trastuzumab-containing neoadjuvant therapy. Methods: We did this multicentre, open-label, randomised trial in women with HER2-positive locally advanced or inflammatory breast cancer. Participants were randomly assigned (1:1), by computer program with a minimisation technique, to receive neoadjuvant chemotherapy alone or with 1 year of trastuzumab (concurrently with neoadjuvant chemotherapy and continued after surgery). A parallel group with HER2-negative disease was included and received neoadjuvant chemotherapy alone. Our primary endpoint was event-free survival. Analysis was by intention to treat. This study is registered at www.controlled-trials.com, ISRCTN86043495. Findings: Between June 20, 2002, and Dec 12, 2005, we enrolled 235 patients with HER2-positive disease, of whom 118 received chemotherapy alone and 117 received chemotherapy plus trastuzumab. 99 additional patients with HER2-negative disease were included in the parallel cohort. After a median follow-up of 5·4 years (IQR 3·1–6·8) the event-free-survival benefit from the addition of trastuzumab to chemotherapy was maintained in patients with HER2-positive disease. 5 year event-free survival was 58% (95% CI 48–66) in patients in the trastuzumab group and 43% (34–52) in those in the chemotherapy group; the unadjusted hazard ratio (HR) for event-free survival between the two randomised HER2-positive treatment groups was 0·64 (95% CI 0·44–0·93; two-sided log-rank p=0·016). Event-free survival was strongly associated with pathological complete remission in patients given trastuzumab. Of the 68 patients with a pathological complete response (45 with trastuzumab and 23 with chemotherapy alone), the HR for event-free survival between those with and without trastuzumab was 0·29 (95% CI 0·11–0·78). During follow-up only four cardiovascular adverse events were regarded by the investigator to be drug-related (grade 2 lymphostasis and grade 2 lymphoedema, each in one patient in the trastuzumab group, and grade 2 thrombosis and grade 2 deep vein thrombosis, each in one patient in the chemotherapy-alone group). Interpretation: These results show a sustained benefit in event-free survival from trastuzumab-containing neoadjuvant therapy followed by adjuvant trastuzumab in patients with locally advanced or inflammatory breast cancer, and provide new insight into the association between pathological complete remission and long-term outcomes in HER2-positive disease. Funding: F Hoffmann-La Roche. [ABSTRACT FROM AUTHOR]

Abstract: A long course of anticancer therapy may lead to testicular steroidogenesis destruction. Cryopreservation of testicular interstitial cells (TIC) would be a strategy to protect hormonal and fertile potential of pre-pubertal boys treated with chemo – or radiotherapy. The aim of this research was to optimize protocols for freezing of TIC. Essential physical processes associated with the presence of dimethyl sulphoxide (Me2SO) in the cryoprotectant solution take place at the temperatures below −60°С. These processes are the eutectic crystallization at the stage of freezing and the recrystallization before the melting of the eutectic mixture at the stage of heating. Both of the processes affect the viability of the cells subjected to cryopreservation. Temperature intervals when these processes take place were determined by the method of thermoplastic deformation for 10% Me2SO selected for cryopreservation of TIC. Rat TIC were cryopreserved using five different protocols which varied in cooling rates within the chosen temperature intervals. Post-thaw cell viability and metabolic activity were evaluated by Trypan Blue and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) staining assays. Leydig cell recovery after cryopreservation was measured by 3-beta-hydroxysteroid dehydrogenase reaction. Based on the obtained results, the authors developed a cryopreservation protocol for TIC which makes it possible to achieve great cell viability due to using controlled cooling rates within the temperature intervals below −60°С. [Copyright &y& Elsevier]

The article discusses a study which assessed event-free survival in patients with HER2-positive locally advanced or inflammatory breast cancer taking neoadjuvant chemotherapy with or without 1 year of trastuzumab. The study involved women treated with trastuzumab and a neoadjuvant chemotherapy regimen comprised of several drugs such as doxorubicin and paclitaxel. The researchers found that there had been an improvement in event-free survival in patients with trastuzumab versus without. They also suggest that the addition of neoadjuvant to chemotherapy be considered.

Background: Few studies have compared primary neoadjuvant endocrine therapy with neoadjuvant chemotherapy in breast cancer patients. The need for preoperative chemotherapy with doxorubicin or taxanes may be reduced in postmenopausal patients with estrogen receptor (ER)-positive and/or progesterone receptor (PgR)-positive tumors. This randomized, controlled, phase 2 study evaluated the efficacy of neoadjuvant chemotherapy compared with endocrine treatment with aromatase inhibitors in postmenopausal women with ER-positive and/or PgR-positive breast cancer.Methods: Eligible patients were randomly assigned to receive neoadjuvant anastrozole 1 mg/day (n = 61) or exemestane 25 mg/day (n = 60) for 3 months or doxorubicin 60 mg/m(2) with paclitaxel 200 mg/m(2) (four 3-week cycles). Study end points included overall objective response determined by palpation, mammography, and ultrasound, and the number of patients who qualified for breast-conserving surgery and radiotherapy.Results: Clinical objective response was 64% in the endocrine therapy and chemotherapy treatment groups. Median time to clinical response was 57 and 51 days with aromatase inhibitors and chemotherapy, respectively (P > .05). Rates of pathological complete response (3% vs 6%) and disease progression (9% vs 9%) did not differ significantly in the endocrine therapy or chemotherapy group, respectively (P > .05). Rates of breast-conserving surgery were slightly higher in the endocrine group (33% vs 24%; P = .058). The most frequent toxicities from chemotherapy were alopecia (79%), grade 3/4 neutropenia (33%), and grade 2 neuropathy (30%). Endocrine treatment was well tolerated. No deaths occurred during the preoperative treatment.Conclusions: Preoperative neoadjuvant endocrine therapy with aromatase inhibitors was well tolerated and resulted in rates similar to chemotherapy in overall objective response and breast-conserving surgery in postmenopausal women with ER-positive and/or PgR-positive tumors. [ABSTRACT FROM AUTHOR]

Few studies have compared primary neoadjuvant endocrine therapy with neoadjuvant chemotherapy in breast cancer patients. The need for preoperative chemotherapy with doxorubicin or taxanes may be reduced in postmenopausal patients with estrogen receptor (ER)‐positive and/or progesterone receptor (PgR)‐positive tumors. This randomized, controlled, phase 2 study evaluated the efficacy of neoadjuvant chemotherapy compared with endocrine treatment with aromatase inhibitors in postmenopausal women with ER‐positive and/or PgR‐positive breast cancer.Eligible patients were randomly assigned to receive neoadjuvant anastrozole 1 mg/day (n = 61) or exemestane 25 mg/day (n = 60) for 3 months or doxorubicin 60 mg/m2 with paclitaxel 200 mg/m2 (four 3‐week cycles). Study end points included overall objective response determined by palpation, mammography, and ultrasound, and the number of patients who qualified for breast‐conserving surgery and radiotherapy.Clinical objective response was 64% in the endocrine therapy and chemotherapy treatment groups. Median time to clinical response was 57 and 51 days with aromatase inhibitors and chemotherapy, respectively (P > .05). Rates of pathological complete response (3% vs 6%) and disease progression (9% vs 9%) did not differ significantly in the endocrine therapy or chemotherapy group, respectively (P > .05). Rates of breast‐conserving surgery were slightly higher in the endocrine group (33% vs 24%; P = .058). The most frequent toxicities from chemotherapy were alopecia (79%), grade 3/4 neutropenia (33%), and grade 2 neuropathy (30%). Endocrine treatment was well tolerated. No deaths occurred during the preoperative treatment.Preoperative neoadjuvant endocrine therapy with aromatase inhibitors was well tolerated and resulted in rates similar to chemotherapy in overall objective response and breast‐conserving surgery in postmenopausal women with ER‐positive and/or PgR‐positive tumors. Cancer 2007. © 2007 American Cancer Society. [ABSTRACT FROM AUTHOR]