EXPERIMENTAL MODELING OF APOPTOSIS UNDER CONDITIONS OF STABLE STRONTIUM EXPOSURE

Apoptosis is defined as a highly regulated form of programmed cell death with typical morphological and biochemical features. A variety of factors, including heavy metals, may influence the intensity of programmed cell death. The aim of the work was to simulate apoptosis in an in vitrosystem under the conditions of stable strontium exposure. The children’s population consuming drinking water with high strontium (Sr2+) content (n = 49) was observed. The level of lymphocyte apoptosis was determined with flow cytometry technique, by means of labeled annexin V-FITC conjugate (AnnV-FITC) and propidium iodide (PI) staining. AnnV-FITC+PI- cells were regarded as early apoptotic forms, whereas late apoptotic and/or necrotic cells were AnnV-FITC+PI+. The isolated leukocytes were incubated with Sr2+ at a concentration of 7.0 mg/l, the maximal permitted concentration (MPC) for water of aqueous objects, for 4 hours at 37 ºC. Expression of CD95 and p53 apoptosis markers was performed by flow cytometry using labeled monoclonal antibodies.In vitroexposure to strontium was associated with significantly decreased expression of apoptosisregulating factors, i.e., membrane marker CD95 and intracellular transcription protein p53, 1.56- and 1.68-fold, respectively. Meanwhile, we revealed a significantly (4.68-fold) decreased amounts of AnnV-FITC+PI--cells, as well as a statistically significant (1.35-fold) increase of the AnnV-FITC+PI+-cells. Moreover, the amounts of AnnV-FITC+ PI--lymphocytes in all samples were below the physiological ranges and control values. The number of samples with higher contents of AnnV-FITC+PI+-lymphocyte exceeding the established standards and control values, was 30.8%. Thus, it has been experimentally proven that strontium, at a concentration corresponding to MPC for water objects may significantly inhibit cell death along apoptotic pathways, with switching to necrotic cell death mechanisms, according to phosphatidylserine contents, as detected by annexin V binding test. The data have revealed an ability of strontium to have a significant effect upon the parameters of regulation and maintenance of cellular homeostasis, by influencing the apoptosis intensity, due to shifting a balance towards necrosis and reducing expression of apoptosis-regulating factors. The results of this study may be used in order to identify some marker indexes of immune disorders potentially induced by external influence of strontium upon human health under specific environmental factors

Биопсия сторожевого лимфатического узла при раке молочной железы с применением метода флуоресцентной визуализации красителя индоцианин зеленый

We presented the preliminary results of sentinel lymph node biopsy (SLN) using the method of fluorescent visualization with the indocyanine green. The study included 40 breast cancer patients with the prevalence of the cTis-2N0M0 process. 1 ml of Indocyanine Green (ICG) aqueous solution was administered periareolarly and subcutaneously from the tumor side with total drug dose of 5 mg. In the following few minutes the ICG trail along the lymphatic vessels was observed by its fluorescence in the infrared spectral range using a special camera with image transmitted to a computer screen. After the trail reached the axillary region and broke off, skin and subcutaneous tissue incision in axillary area was made, and the superficial fascia was dissected. The first contrasted lymph nodes were extracted. The incidence of SLN was 92.5%. Metastases were detected in 20% of cases. On average, it took 17.6 minutes to identify and remove the SLN. In 7 patients the use of the fluorescent SLN detection method was combined with radioisotope (Technefite 99m Tc colloid) – in all cases the same SLN was identified. The use of the ICG lymphotropic dye with the subsequent detection of SLN by the fluorescence method makes it possible to diagnose the status of SLN with a high degree of accuracy and can be comparable in effectiveness with the isotopic method. The specificities of lymphatic drainage were found out with the use of fluorescence lymphography: inverse dependence of the lymphatic drainage rate on the body mass index and breast size, and absence of dependence on degree of ptosis of breast.В работе представлены результаты исследования биопсии сторожевого лимфатического узла (СЛУ) с помощью метода флуоресцентной визуализации красителя индоцианин зеленый. В исследование были включены 40 больных раком молочной железы с распространенностью процесса сTis-2N0M0. 1 мл водного раствора красителя индоцианина зеленый вводили периареолярно внутрикожно и подкожно со стороны опухоли в дозе активного вещества 5 мг. В ближайшие минуты наблюдали движение красителя по лимфатическим путям в виде «дорожки» способом флуоресценции в инфракрасном спектре с помощью специальной камеры с передачей изображения на экран компьютера. После того, как дорожка достигала подмышечной области и обрывалась, производили разрез кожи и подкожной клетчатки в подмышечной области, рассекали поверхностную фасцию. Выделяли первые контрастиро ванные лимфатические узлы. Частота выявления СЛУ составила 92,5%, из них метастазы выявлены в 20% случаях. В среднем на этап идентификации и удаления СЛУ уходило 17,6 мин. У 7 больных комбинировали применение метода флуоресцентного (краситель ICG) определения СЛУ с радиоизотопным (коллоид Технефит 99m Tc) – во всех случаях идентифицировали одни и те же СЛУ. Применение лимфотропного красителя индоцианина зеленого с последующим выявлением СЛУ методом флуоресценции позволяет с высокой степенью точности диагностировать состояние СЛУ, а также сопоставимо по эффективности с изотопным методом. Выявлены осо бенности лимфооттока в молочной железе с помощью флуоресцентной лимфографии: обратная зависимость скорости лимфоотто ка от индекса массы тела и размера молочной железы, отсутствие зависимости от степени птоза железы

Directional Sensitivity of the NEWSdm Experiment to Cosmic Ray Boosted Dark Matter

We present a study of a directional search for Dark Matter boosted forward when scattered by cosmic-ray nuclei, using a module of the NEWSdm experiment. The boosted Dark Matter flux at the edge of the Earth's atmosphere is expected to be pointing to the Galactic Center, with a flux 15 to 20 times larger than in the transverse direction. The module of the NEWSdm experiment consists of a 10 kg stack of Nano Imaging Trackers, i.e.~newly developed nuclear emulsions with AgBr crystal sizes down to a few tens of nanometers. The module is installed on an equatorial telescope. The relatively long recoil tracks induced by boosted Dark Matter, combined with the nanometric granularity of the emulsion, result in an extremely low background. This makes an installation at the INFN Gran Sasso laboratory, both on the surface and underground, viable. A comparison between the two locations is made. The angular distribution of nuclear recoils induced by boosted Dark Matter in the emulsion films at the surface laboratory is expected to show an excess with a factor of 3.5 in the direction of the Galactic Center. This excess allows for a Dark Matter search with directional sensitivity. The surface laboratory configuration prevents the deterioration of the signal in the rock overburden and it emerges as the most powerful approach for a directional observation of boosted Dark Matter with high sensitivity. We show that, with this approach, a 10 kg module of the NEWSdm experiment exposed for one year at the Gran Sasso surface laboratory can probe Dark Matter masses between 1 keV/c$^2$ and 1 GeV/c$^2$ and cross-section values down to $10^{-30}$~cm$^2$ with a directional sensitive search.Comment: 15 pages, 14 figures, updated references, clarified discussion in intro section. Submitted to JCA

SARS Coronavirus 3b Accessory Protein Modulates Transcriptional Activity of RUNX1b

BACKGROUND: The causative agent of severe acute respiratory syndrome, SARS coronavirus (SARS-CoV) genome encodes several unique group specific accessory proteins with unknown functions. Among them, accessory protein 3b (also known as ORF4) was lately identified as one of the viral interferon antagonist. Recently our lab uncovered a new role for 3b in upregulation of AP-1 transcriptional activity and its downstream genes. Thus, we believe that 3b might play an important role in SARS-CoV pathogenesis and therefore is of considerable interest. The current study aims at identifying novel host cellular interactors of the 3b protein. METHODOLOGY/PRINCIPAL FINDINGS: In this study, using yeast two-hybrid and co-immunoprecipitation techniques, we have identified a host transcription factor RUNX1b (Runt related transcription factor, isoform b) as a novel interacting partner for SARS-CoV 3b protein. Chromatin immunoprecipitaion (ChIP) and reporter gene assays in 3b expressing jurkat cells showed recruitment of 3b on the RUNX1 binding element that led to an increase in RUNX1b transactivation potential on the IL2 promoter. Kinase assay and pharmacological inhibitor treatment implied that 3b also affect RUNX1b transcriptional activity by regulating its ERK dependent phosphorylation levels. Additionally, mRNA levels of MIP-1α, a RUNX1b target gene upregulated in SARS-CoV infected monocyte-derived dendritic cells, were found to be elevated in 3b expressing U937 monocyte cells. CONCLUSIONS/SIGNIFICANCE: These results unveil a novel interaction of SARS-CoV 3b with the host factor, RUNX1b, and speculate its physiological relevance in upregulating cytokines and chemokine levels in state of SARS virus infection

Sentinel lymph node biopsy for breast cancer using indocyanine green fluorescence visualization

We presented the preliminary results of sentinel lymph node biopsy (SLN) using the method of fluorescent visualization with the indocyanine green. The study included 40 breast cancer patients with the prevalence of the cTis-2N0M0 process. 1 ml of Indocyanine Green (ICG) aqueous solution was administered periareolarly and subcutaneously from the tumor side with total drug dose of 5 mg. In the following few minutes the ICG trail along the lymphatic vessels was observed by its fluorescence in the infrared spectral range using a special camera with image transmitted to a computer screen. After the trail reached the axillary region and broke off, skin and subcutaneous tissue incision in axillary area was made, and the superficial fascia was dissected. The first contrasted lymph nodes were extracted. The incidence of SLN was 92.5%. Metastases were detected in 20% of cases. On average, it took 17.6 minutes to identify and remove the SLN. In 7 patients the use of the fluorescent SLN detection method was combined with radioisotope (Technefite 99m Tc colloid) – in all cases the same SLN was identified. The use of the ICG lymphotropic dye with the subsequent detection of SLN by the fluorescence method makes it possible to diagnose the status of SLN with a high degree of accuracy and can be comparable in effectiveness with the isotopic method. The specificities of lymphatic drainage were found out with the use of fluorescence lymphography: inverse dependence of the lymphatic drainage rate on the body mass index and breast size, and absence of dependence on degree of ptosis of breast

Optimization of dossiers content in compositional materials by experimental and statistical modeling methods

Проведено дослідження композиційного матеріалу, до складу якого входять алюмосилікатні мікросфери, портландцемент, добавка метилцелюлози та комплексна хімічна добавка, що сприяє додатковому синтезу кристалогідратів з алюмінатними та силікатними фазами цементу. В роботі було поставлено задачу, а саме оптимізувати вміст складових композиту за допомогою математичного моделювання. Було отримано модель у вигляді поліному другого порядку. Перевірено значимість коефіцієнтів у рівнянні, адекватність моделі. Побудовано поверхні відгуку та їх проекції на площину. Внаслідок чого було отримано скорегований склад і раціональні області вмісту складових суміші із застосуванням математичного апарату.иционного материала, в состав которого входят алюмосиликатные микросферы, ы и комплексная химическая добавка, которая способствует дополнительно с алюминатными и силикатными фазами цемента. В работе была поставлена задача, а овать содержание составляющих композита с помощью математического моделирования. Была Проверена значимость коэффициентов в уравнении, адекватность на плоскость. В результате был получен скорректированный жания составляющих смеси с применением математического аппарата. ные и стеклянные микросферы, глубина проникновения, матрица планирования, натуральные переменные. SSIERS CONTENT IN COMPOSITIONAL MATERIALS TAL AND STATISTICAL MODELING METHODS K. Plakhotnikov, O. Starkova, T. Kostyuk, D. Bondarenko tent of methylcellulose and complex chemical additives in the composition of a is of Portland cement is considered in order to achieve maximum depth of penetration of a composite fluid into a structure of concrete or other mineral substrates. Dos-tracking of composite material consisting of aluminosilicate microspheres, portland cement, methylcellulose addit and complex chemical additive, which contributes to the additional s he microlevel is invest statistical methods of planning the experimen e using the tools of the package S ОПТИМИЗАЦИЯ СОДЕРЖАНИЯ ДО МЕТОДАМИ ЭКСПЕРИМЕНТАЛЬ К.В. П Проведено исследование композ етилцеллюлозпортландцемент, добавка м у синтезу кристаллогидратовм именно оптимизир получена модель в виде полинома второго порядка. модели. Построены поверхности отклика и их проекции ьные области содерсостав и рационалThe possibility of obtaining a rational content of methylcellulose and complex chemical additives in the composition of a heat-insulating composite on the basis of Portland cement is considered in order to achieve maximum depth of penetration of a composite fluid into a structure of concrete or other mineral substrates. Dos-tracking of composite material consisting of aluminosilicate microspheres, portland cement, methylcellulose additive and complex chemical additive, which contributes to the additional synthesis of crystalline hydrates with aluminate and silicate phases of cement. The structure of the composite at the microlevel is investigated by means of electron microscopy. Rational content of supplements was carried out using statistical methods of planning the experiment, obtaining a mathematical model and constructing a response surface using the tools of the package Statistica 6.0. The quantitative selection of a complex chemical additive can further synthesize on the surface of microspheres crystalline hydrates in the form of gel aggregates CSH, fibers of etringitis and laminar C4AH13. With excessive amounts of methylcellulose additives and the lack of a complex chemical additive, the integrity of the crystalline shell on the surface of the microspheres is violated and the penetration depth of the substrate applied to the porous (concrete, brick, etc.) composite material is reduced, which reduces its adhesion to the base. The most rational is the value when the content of methylcellulose and complex chemical additives is maximal or minimal, within the limits set by search experiments, but taking into account the economic component, it is advisable to choose the values in the minimal limits with the most rational one