In vitro culture of primary human myoblasts by using the dextran microcarriers Cytodex3®

Introduction. Primary cells in vitro culture scale-up is a crucial issue in cell-based tissue and organ regeneration therapy. Reducing costs and space occupied by the cells cultured in vitro has been an important target. Cells cultured in vitro with the use of bioreactor with dextran microcarriers (Cytodex®) have potentially a chance to meet many of the cell therapy requirements. Material and methods. We used collagen-coated carriers (Cytodex3®) and a spinner flask bioreactor to develop environment suitable for human myoblast proliferation. In parallel, standard adherent in vitro culture conditions for myoblasts propagation (T-flask) were conducted. Cell cycle characterization, senescence, myogenic gene ex­pression and cell apoptosis were evaluated in order to find differences between two culture systems under study. Results. The number of cells obtained in bioreactor per 106 of starting cells population was approximately ten times lower in comparison with T-flask culture system. The microcarriers cultured adult myoblasts in compari­son with the regular T-flask culture showed faster and more advanced replicative aging and lower proliferative potential. Moreover, the percentage of the cells that entailed an irreversible cell arrest (G0 phase) was also significantly (p < 0.0001) increased. Conclusions. Our results suggest that population of primary human myoblasts obtained from adult individuals and propagated on dextran microcarriers did not meet the requirements of the regenerative medicine regarding quantity and quality of the cells obtained. Nonetheless, further optimization of the cell scaling up process including both microcarriers and/or bioreactor program is still an important option

The potential impact of hereditary hemochromatosis on the heart considering the disease stage and patient age—the role of echocardiography

BackgroundHereditary hemochromatosis (HH) is a genetic disease that leads to increased iron accumulation in several organs. Cardiomyocytes are highly susceptible to this damage owing to their high iron uptake, and cardiovascular complications account for 1/3 of the deaths in the natural course of HH. Additionally, excess iron intake and associated oxidative stress may accelerate the aging of the cardiovascular system, regardless of the age of patients with HH. We aimed to investigate the role of standard and speckle-tracking echocardiography (STE) in revealing heart differences in patients with HH considering the disease stage and the patient age.MethodologyConsecutive patients with HH (n = 58) without heart pathologies (except hypertension) and 29 age- and sex-matched healthy individuals underwent echocardiography. Patients were compared according to the time since HH diagnosis (the recently diagnosed HH group [31 patients] with diagnosed HH for less than 6 months and had no more than one venesection; the medium group [11 patients] with diagnosed HH between 6 and 24 months; and the long-lasting group [16 patients] with diagnosed HH for more than 2 years) and the quartile contribution of their age.ResultsStandard echocardiography revealed differences in diastolic parameters between patients with HH and controls, which were the most prominent between healthy and long-lasting HH patients. Regarding systolic function, left ventricular ejection fraction was lower in HH patients, with the most evident differences between the healthy and recently diagnosed HH patients. STE revealed additional differences in systolic parameters, with LV rotation the worst in recently diagnosed patients and its increase in patients with medium and long-lasting HH. Significantly worse peak systolic longitudinal strain values were observed in all patients with HH. Analyses of the results according to the age quartiles of patients with HH revealed that some changes ocurred earlier than expected according to age.ConclusionsEchocardiography can reveal possible heart damage in HH patients at different stages of the disease and highlight potential features of accelerated myocardial aging in these patients

Nadekspresja miR-652-5p u chorych z nowo rozpoznaną cukrzycą typu 1

Wstęp. MicroRNA (miRNA) to krótkie, niekodujące RNA, które regulują ekspresję genów na poziomie potransktypcyjnym. Cząsteczki miRNA odgrywają istotną rolę w regulacji centralnej i obwodowej tolerancji immunologicznej, dlatego analiza niekodujących RNA w cukrzycy typu 1 (T1D) jest istotnym problemem badawczym. Celem badania była analiza ekspresji miR-652-5p w nowo zdiagnozowanych przypadkach cukrzycy typu 1 oraz ocena miejsc wiązania miR-652-5p w potencjalnych genach docelowych ADAR i MARCH5. Metody. Analiza ekspresji miR-652-5p została przeprowadzona w komórkach jednojądrzastych krwi obwodowej dzieci z nowo rozpoznaną cukrzycą typu 1 (n = 28) oraz w dobranej pod względem wieku grupie kontrolnej (n = 28) na podstawie metody PCR w czasie rzeczywistym. Miejsca wiązania miR-652-5p w genach docelowych zostały ocenione z wykorzystaniem układu reporterowego lucyferazy. Wyniki. Analiza ekspresji wykazała podwyższony poziom miR-652-5p u pacjentów z T1D w porównaniu z grupą zdrowych dawców (p < 0,05). Ocena miejsc wiązania w układzie reporterowym lucyferazy nie wskazała genu ADAR i MARCH5 jako potencjalnych genów docelowych miR-652-5p. Wnioski. Przeprowadzone badania wskazały miR-652-5p jako potencjalny biomarker cukrzycy typu 1

Does social media presence of luxury brands affect consumers´ perceived luxury value?

Social media has fundamentally transformed the business environment. In fact, in most industries social media adoption has been relatively rapid, however, luxury brands’ relationship with social media is slightly more complicated. The appeal of luxury products stems from their prestige and exclusivity. This is not the case for social media platforms. They pose both a challenge and an opportunity for luxury sector. Hence objective of this study is to investigate whether social media presence of luxury brands affects consumers’ perceived luxury value. Through an experimental research design and quantitative data collection, results show that the former has little to no effect on the latter

Does the age of patients with hereditary hemochromatosis at the moment of their first diagnosis have an additional effect on the standard echocardiographic parameters?

Background: Hereditary haemochromatosis (HH) is an inherited disease in which gene mutation leads to excessive iron absorption and accumulation in different organs, including the heart, which causes damage. Whether the age of patients with HH at the moment of their first diagnosis has an additional effect on the standard echocardiographic parameters was the aim of the study. Material and methods: We prospectively enrolled 20 HH patients, and 20 healthy age- and sex-matched volunteers. Analysis of standard echocardiographic parameters was performed and compared in subgroups of ≥50 and <50 years old (yo). Results: Comparing HH patients with healthy volunteers in ≥50 yo subgroup, significant differences were found in parameters regarding diastolic function (IVS thickness, LVM index, Em, E/Em, PV S/D, LAA index and LAV index). In the <50 yo subgroup we did not find the abovementioned differences, however LVEF appeared to be lower in the HH patients. Conclusions: Despite the lack of clinical symptoms of cardiovascular disease and the lack of deviations in the standard echocardiographic examination, there were a number of differences regarding LV diastolic function parameters in HH patients ≥50 yo, whereas differences regarding LV systolic function were more prominent in HH patients <50 yo when compared with healthy subjects

Autonomic Predictors of Hospitalization Due to Heart Failure Decompensation in Patients with Left Ventricular Systolic Dysfunction.

INTRODUCTION:Autonomic nervous system balance can be significantly deteriorated during heart failure exacerbation. However, it is still unknown whether these changes are only the consequence of heart failure decompensation or can also predict development thereof. Objectives were to verify if simple, non-invasive autonomic parameters, such as baroreflex sensitivity and short-term heart rate variability can provide independent of other well-known clinical parameters information on the risk of heart failure decompensation in patients with left ventricular systolic dysfunction. METHODS:In 142 stable patients with left ventricular ejection fraction ≤ 40%, baroreflex sensitivity and short-term heart rate variability, as well as other well-known clinical parameters, were analyzed. During 23 ± 9 months of follow-up 19 patients were hospitalized due to the heart failure decompensation (EVENT). RESULTS:Pre-specified cut-off values of baroreflex sensitivity (≤2.4 ms/mmHg) and low frequency power index of heart rate variability (≤19 ms2) were significantly associated with the EVENTs (hazard ratio 4.43, 95% confidence interval [CI] 1.35-14.54 and 5.41, 95% CI 1.87-15.65 respectively). EVENTs were also associated with other parameters, such as left ventricular ejection fraction, NYHA class, diuretic use, renal function, brain natriuretic peptide and hemoglobin level, left atrial size, left and right ventricular heart failure signs. After adjusting baroreflex sensitivity and low frequency power index for each of the abovementioned parameters, autonomic parameters were still significant predictors of hospitalization due to the heart failure decompensation. CONCLUSION:Simple, noninvasive autonomic indices can be helpful in identifying individuals with increased risk of hospitalization due to the heart failure decompensation among clinically stable patients with left ventricular systolic dysfunction, even when adjusted for other well-known clinical parameters

Quality of life and satisfaction with life of malaria patients in context of acceptance of the disease: quantitative studies

Abstract Background Health status is one of the basic factors of a high quality of life and the problem of the acceptance of illness is important for adaptation to the limitations imposed by it. The purpose of the study was the evaluation of the quality of life, satisfaction with life and the acceptance of illness by malaria patients, as well as the discovery of a relationship between studied parameters. Methods The study was undertaken in August 2010, on 120 Nigerian patients with confirmed malaria. A method of diagnostic survey, based on standardized scales - Acceptance of Illness Scale, The Satisfaction With Life Scale and a standardized survey questionnaire World Health Organization Quality of Life/BREF - was used in this study. Descriptive statistics, variability range, 95% confidence interval, correlation analysis, Spearman’s non-parametric correlation coefficient, Mann–Whitney test and Kruskal-Wallis test were applied and the, so called, test statistics was calculated, followed by the calculation of the test probability p. Results of analyses were presented in a box graph, and a graph of dispersion. Results A dominating share in the adjective scale of the AIS scale was the category of “no acceptance”, given by 71.7% of respondents. The average level of a “somatic domain” was 41.7, and of a “social domain” was 62.8. The mean satisfaction of life evaluation in the SWLS scale was 18 points. The correlation between acceptance of the disease and quality of life for the psychological domain was 0.39***, and between acceptance of the disease and satisfaction with life was 0.40***. The correlation between satisfaction with life and quality of life for the psychological domain was 0.65***, and between satisfaction with life and quality of life for the environment domain was 0.60***. The mean level of AIS for the studied population of men was 16.5, and test probability: p = 0.0014**, and for the environment domain the level was 50, and the test probability: p = 0.0073**. For quality of life in the social sphere the test probability: p = 0.0013** in relatively older individuals. Conclusion The majority of people do not accept their condition. Evaluation of the quality of life was the highest in the social domain, and the lowest in the somatic domain. There is a statistically significant correlation between the level of acceptance of illness and the quality of life and satisfaction with life. The strongest correlation is found between satisfaction with life and the evaluation of the quality of life in psychological and environmental domains. Men evaluate their quality of life in the environmental domain higher and demonstrate a higher acceptance of their disease. There is a correlation regarding a significantly higher quality of life in the social sphere in relatively older people.</p

pNiPAM-Nanoparticle-Based Antiapoptotic Approach for Pro-Regenerative Capacity of Skeletal Myogenic Cells

The biocompatibility of pNiPAM (Poly N-isopropylacrylamide) copolymers has been examined and they did not exert any cytotoxic effects. Their properties and vulnerable temperature characteristics make them candidates for use in medical applications. We synthesized a well-characterized nanoparticles-based cargo system that would effectively deliver a biological agent to human skeletal myogenic cells (SkMCs); among other aspects, a downregulating apoptotic pathway potentially responsible for poor regeneration of myocardium. We confirmed the size of the pNiPAM based spheres at around 100 nm and the nanomeric shape of nanoparticles (NP) obtained. We confirmed that 33 °C is the adequate temperature for phase transition. We performed the dynamics of cargo release. A small amount of examined protein was detected at 10 min after reaching LCTS (lower critical solution temperature). The presented results of the test with BSA (bovine serum albumin) and doxorubicin loaded into nanoparticles showed a similar release profile for both substances. SkMCs incubated with NP loaded with antiapoptotic agent, BCB (Bax channel blocker), significantly diminished cell apoptosis (p &lt; 0.01). Moreover, the lowest apoptotic level was detected in SkMCs treated with camptothecin and simultaneously incubated with pNiPAMs loaded with BCB. Application of nanoparticles loaded with BCB or subjected to BCB alone did not, however, diminish the amount of apparently necrotic cells

Quality of life and satisfaction with life of malaria patients in context of acceptance of the disease: quantitative studies

BACKGROUND: Health status is one of the basic factors of a high quality of life and the problem of the acceptance of illness is important for adaptation to the limitations imposed by it. The purpose of the study was the evaluation of the quality of life, satisfaction with life and the acceptance of illness by malaria patients, as well as the discovery of a relationship between studied parameters. METHODS: The study was undertaken in August 2010, on 120 Nigerian patients with confirmed malaria. A method of diagnostic survey, based on standardized scales - Acceptance of Illness Scale, The Satisfaction With Life Scale and a standardized survey questionnaire World Health Organization Quality of Life/BREF - was used in this study. Descriptive statistics, variability range, 95% confidence interval, correlation analysis, Spearman’s non-parametric correlation coefficient, Mann–Whitney test and Kruskal-Wallis test were applied and the, so called, test statistics was calculated, followed by the calculation of the test probability p. Results of analyses were presented in a box graph, and a graph of dispersion. RESULTS: A dominating share in the adjective scale of the AIS scale was the category of “no acceptance”, given by 71.7% of respondents. The average level of a “somatic domain” was 41.7, and of a “social domain” was 62.8. The mean satisfaction of life evaluation in the SWLS scale was 18 points. The correlation between acceptance of the disease and quality of life for the psychological domain was 0.39***, and between acceptance of the disease and satisfaction with life was 0.40***. The correlation between satisfaction with life and quality of life for the psychological domain was 0.65***, and between satisfaction with life and quality of life for the environment domain was 0.60***. The mean level of AIS for the studied population of men was 16.5, and test probability: p = 0.0014**, and for the environment domain the level was 50, and the test probability: p = 0.0073**. For quality of life in the social sphere the test probability: p = 0.0013** in relatively older individuals. CONCLUSION: The majority of people do not accept their condition. Evaluation of the quality of life was the highest in the social domain, and the lowest in the somatic domain. There is a statistically significant correlation between the level of acceptance of illness and the quality of life and satisfaction with life. The strongest correlation is found between satisfaction with life and the evaluation of the quality of life in psychological and environmental domains. Men evaluate their quality of life in the environmental domain higher and demonstrate a higher acceptance of their disease. There is a correlation regarding a significantly higher quality of life in the social sphere in relatively older people

Hypoxia-Induced FAM13A Regulates the Proliferation and Metastasis of Non-Small Cell Lung Cancer Cells

Hypoxia in non-small cell lung cancer (NSCLC) affects cancer progression, metastasis and metabolism. We previously showed that FAM13A was induced by hypoxia in NSCLC but the biological function of this gene has not been fully elucidated. This study aimed to investigate the role of hypoxia-induced FAM13A in NSCLC progression and metastasis. Lentiviral shRNAs were used for FAM13A gene silencing in NSCLC cell lines (A549, CORL-105). MTS assay, cell tracking VPD540 dye, wound healing assay, invasion assay, BrdU assay and APC Annexin V staining assays were performed to examine cell proliferation ability, migration, invasion and apoptosis rate in NSCLC cells. The results of VPD540 dye and MTS assays showed a significant reduction in cell proliferation after FAM13A knockdown in A549 cells cultured under normal and hypoxia (1% O2) conditions (p &lt; 0.05), while the effect of FAM13A downregulation on CORL-105 cells was observed after 96 h exposition to hypoxia. Moreover, FAM13A inhibition induced S phase cell cycle arrest in A549 cells under hypoxia conditions. Silencing of FAM13A significantly suppressed migration of A549 and CORL-105 cells in both oxygen conditions, especially after 72 and 96 h (p &lt; 0.001 in normoxia, p &lt; 0.01 after hypoxia). It was showed that FAM13A reduction resulted in disruption of the F-actin cytoskeleton altering A549 cell migration. Cell invasion rates were significantly decreased in A549 FAM13A depleted cells compared to controls (p &lt; 0.05), mostly under hypoxia. FAM13A silencing had no effect on apoptosis induction in NSCLC cells. In the present study, we found that FAM13A silencing has a negative effect on proliferation, migration and invasion activity in NSCLC cells in normal and hypoxic conditions. Our data demonstrated that FAM13A depleted post-hypoxic cells have a decreased cell proliferation ability and metastatic potential, which indicates FAM13A as a potential therapeutic target in lung cancer