Search results for: Mikolaj Wrobel

Authors: Katarzyna Drela, Miroslaw Wielgos, Mikolaj Wrobel, Barbara Lukomska

Abstract:

Mesenchymal stem cells (MSCs) have a great potential in regenerative medicine. Since the initial number of isolated MSCs is limited, in vitro propagation is often required to reach sufficient numbers of cells for therapeutic applications. During long-term culture MSCs may undergo genetic or epigenetic alterations that subsequently increase the probability of spontaneous malignant transformation. Thus, factors that influence genomic stability of MSCs following long-term expansions need to be clarified before cultured MSCs are employed for clinical application. The aim of our study was to investigate the potential for spontaneous transformation of human neonatal cord blood (HUCB-MSCs) and adult bone marrow (BM-MSCs) derived MSCs. Materials and Methods: HUCB-MSCs and BM-MSCs were isolated by standard Ficoll gradient centrifugations method. Isolated cells were initially plated in high density 106 cells per cm2. After 48 h medium were changed and non-adherent cells were removed. The malignant transformation of MSCs in vitro was evaluated by morphological changes, proliferation rate, ability to enter cell senescence, the telomerase expression and chromosomal abnormality. Proliferation of MSCs was analyzed with WST-1 reduction method and population doubling time (PDT) was calculated at different culture stages. Then the expression pattern of genes characteristic for mesenchymal or epithelial cells, as well as transcriptions factors were examined by RT-PCR. Concomitantly, immunocytochemical analysis of gene-related proteins was employed. Results: Our studies showed that MSCs from all bone marrow isolations ultimately entered senescence and did not undergo spontaneous malignant transformation. However, HUCB-MSCs from one of the 15 donors displayed an increased proliferation rate, failed to enter senescence, and exhibited an altered cell morphology. In this sample we observed two different cell phenotypes: one mesenchymal-like exhibited spindle shaped morphology and express specific mesenchymal surface markers (CD73, CD90, CD105, CD166) with low proliferation rate, and the second one with round, densely package epithelial-like cells with significantly increased proliferation rate. The PDT of epithelial-like populations was around 1day and 100% of cells were positive for proliferation marker Ki-67. Moreover, HUCB-MSCs showed a positive expression of human telomerase reverse transcriptase (hTERT), cMYC and exhibit increased number of CFU during the long-term culture in vitro. Furthermore, karyotype analysis revealed chromosomal abnormalities including duplications. Conclusions: Our studies demonstrate that HUCB-MSCs are susceptible to spontaneous malignant transformation during long-term culture. Spontaneous malignant transformation process following in vitro culture has enormous effect on the biosafety issues of future cell-based therapies and regenerative medicine regimens.

Keywords: mesenchymal stem cells, spontaneous, transformation, long-term culture

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Authors: Ana Beatriz C. G. Silva, Luiz Carlos Wrobel, Fernando Luiz B. Ribeiro

Abstract:

This paper presents a thermoregulation model using the finite element method to perform numerical analyses of brain cooling procedures as a contribution to the investigation on the use of therapeutic hypothermia after ischemia in adults. The use of computational methods can aid clinicians to observe body temperature using different cooling methods without the need of invasive techniques, and can thus be a valuable tool to assist clinical trials simulating different cooling options that can be used for treatment. In this work, we developed a FEM package applied to the solution of the continuum bioheat Pennes equation. Blood temperature changes were considered using a blood pool approach and a lumped analysis for intravascular catheter method of blood cooling. Some analyses are performed using a three-dimensional mesh based on a complex geometry obtained from computed tomography medical images, considering a cooling blanket and a intravascular catheter. A comparison is made between the results obtained and the effects of each case in brain temperature reduction in a required time, maintenance of body temperature at moderate hypothermia levels and gradual rewarming.

Keywords: brain cooling, finite element method, hypothermia treatment, thermoregulation

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Authors: Luiz Carlos Wrobel, Matjaz Hribersek, Jure Marn, Jurij Iljaz

Abstract:

Dynamic thermography has been clinically proven to be a valuable diagnostic technique for skin tumor detection as well as for other medical applications such as breast cancer diagnostics, diagnostics of vascular diseases, fever screening, dermatological and other applications. Thermography for medical screening can be done in two different ways, observing the temperature response under steady-state conditions (passive or static thermography), and by inducing thermal stresses by cooling or heating the observed tissue and measuring the thermal response during the recovery phase (active or dynamic thermography). The numerical modelling of heat transfer phenomena in biological tissue during dynamic thermography can aid the technique by improving process parameters or by estimating unknown tissue parameters based on measured data. This paper presents a nonlinear numerical model of multilayer skin tissue containing a skin tumor, together with the thermoregulation response of the tissue during the cooling-rewarming processes of dynamic thermography. The model is based on the Pennes bioheat equation and solved numerically by using a subdomain boundary element method which treats the problem as axisymmetric. The paper includes computational tests and numerical results for Clark II and Clark IV tumors, comparing the models using constant and temperature-dependent thermophysical properties, which showed noticeable differences and highlighted the importance of using a local thermoregulation model.

Keywords: boundary element method, dynamic thermography, static thermography, skin tumor diagnostic

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Authors: Sylwia Orzechowska, Andrzej Wróbel, Eugeniusz Rokita

Abstract:

The mineralization is a curious problem of connective tissues. Factors which may play a decisive role in the regulation of the yellow ligaments (YL) mineralization are still open questions. The aim of the studies was a detailed description of the chemical composition and morphology of mineral deposits in the human yellow ligaments. Investigations of the structural features of deposits were used to explain the impact of various factors on mineralization process. The studies were carried out on 24 YL samples, surgically removed from patients suffer from spinal canal stenosis and the patients who sustained a trauma. The micro-computed tomography was used to describe the morphology of mineral deposits. The X-ray fluorescence method and Fourier transform infrared spectroscopy were applied to determine the chemical composition of the samples. In order to eliminate the effect of blur in microtomographic images, the correction method of partial volume effect was used. The mineral deposits appear in 60% of YL samples, both in patients with a stenosis and following injury. The mineral deposits have a heterogeneous structure and they are a mixture of the tissue and mineral grains. The volume of mineral grains amounts to (1.9 ± 3.4)*10-3 mm3 while the density distribution of grains occurs in two distinct ranges (1.75 - 2.15 and 2.15-2.5) g/cm3. Application of the partial volume effect correction allows accurate calculations by eliminating the averaging effect of gray levels in tomographic images. The B-type carbonate-containing hydroxyapatite constitutes the mineral phase of majority YLs. The main phase of two samples was calcium pyrophosphate dihydrate (CPPD). The elemental composition of minerals in all samples is almost identical. This pathology may be independent on the spine diseases and it does not evoke canal stenosis. The two ranges of grains density indicate two stages of grains growth and the degree of maturity. The presence of CPPD crystals may coexist with other pathologies.

Keywords: FTIR, micro-tomography, mineralization, spinal ligaments

Procedia PDF Downloads 357

Authors: Sylwia Laba, Robert Laba, Krystian Szczepanski, Mikolaj Niedek, Anna Kaminska-Dworznicka

Abstract:

Meat and its products are considered food products, having the most unfavorable effect on the environment that requires rational management of these products and waste, originating throughout the whole chain of manufacture, processing, transport, and trade of meat. From the economic and environmental viewpoints, it is important to limit the losses and food wastage and the food waste in the whole meat sector. The link to basic production includes obtaining raw meat, i.e., animal breeding, management, and transport of animals to the slaughterhouse. Food is any substance or product, intended to be consumed by humans. It was determined (for the needs of the present studies) when the raw material is considered as a food. It is the moment when the animals are prepared to loading with the aim to be transported to a slaughterhouse and utilized for food purposes. The aim of the studies was to determine the reasons for loss generation in the basic production of the meat sector in Poland during the years 2017 – 2018. The studies on food losses and waste in the meat sector in basic production were carried out in two areas: red meat i.e., pork and beef and poultry meat. The studies of basic production were conducted in the period of March-May 2019 at the territory of the whole country on a representative trial of 278 farms, including 102 pork production, 55–beef production, and 121 poultry meat production. The surveys were carried out with the utilization of questionnaires by the PAPI (Paper & Pen Personal Interview) method; the pollsters conducted direct questionnaire interviews. Research results indicate that it is followed that any losses were not recorded during the preparation, loading, and transport of the animals to the slaughterhouse in 33% of the visited farms. In the farms where the losses were indicated, the crushing and suffocations, occurring during the production of pigs, beef cattle and poultry, were the main reasons for these losses. They constituted ca. 40% of the reported reasons. The stress generated by loading and transport caused 16 – 17% (depending on the season of the year) of the loss reasons. In the case of poultry production, in 2017, additionally, 10.7% of losses were caused by inappropriate conditions of loading and transportation, while in 2018 – 11.8%. The diseases were one of the reasons for the losses in pork and beef production (7% of the losses). The losses and waste, generated during livestock production and in meat processing and trade cannot be managed or recovered. They have to be disposed of. It is, therefore, important to prevent and minimize the losses throughout the whole production chain. It is possible to introduce the appropriate measures, connected mainly with the appropriate conditions and methods of animal loading and transport.

Keywords: food losses, food waste, livestock production, meat sector

Procedia PDF Downloads 107