Photoprotection in Patients after Kidney Transplantation Author: Zuzana Hajšelová Tutor: doc. PharmDr. Josef Malý, PhD. Consultant: Mgr. Barbora Vaňková Department of Social and Clinical Pharmacy, Faculty of Pharmacy in Hradci Králové, Charles University Introduction and aims: Patients after kidney transplantation (KT) are in a higher risk of developing skin cancer as a result of lifelong immunosuppressive (IS) therapy. Thorough photoprotection is therefore recommended. The aim of this study was to determine patients' awareness and level of photoprotection and to analyse selected risk factors for skin cancer development in patients after KT. Methods: The cross-sectional study was undertaken from 15th April to 31st December 2019 at the Haemodialysis Centre in the Teaching Hospital Hradec Králové. Patients included in this study were above 18 years old, who visited Haemodialysis Centre in the defined period. The data was collected from the patients' medical documentation (e.g. occurrence of skin cancer and skin cancer type, current IS) and from the written version of questionnaires (e.g. phototype, sun exposure, photoprotection). The data was analysed using descriptive statistics. Results: There were 410 KT patients in regular care of the Haemodialysis Centre, 361 fulfilled the questionnaire (88.0...

Stem cell transplantation is a known treatment for various cancers. Currently most cells transplanted are collected via apheresis. An injection of growth factor is given to the patient to start the proliferation and mobilization of stem cells. Apheresis can be initiated when the patient has a stem cell count of 15 to 20 stem cells/µL of peripheral blood. The standard method with which stem cells are analysed is immune flow cytometry where CD34+ and CD45+ are identified with targeted fluorescent antibodies. This analysis takes more than 45 minutes to perform.     Sysmex XN-9000 analyses samples with flow cytometry by lysing erythrocytes and platelets and staining the leukocytes with fluorescent dye. Analysis of the hematopoietic progenitor cells (HPC) takes less than 4 minutes. The purpose of this study was to investigate ifit is possible to predict the start of the apheresis using XN-9000.     For this study, 43 samples were analysed using both methods. Using the sign test, a p-value was calculated to <0.05, which indicates a significant difference between the results received by the two methods. Spearman’s rank correlation gave an observed ρ-value > the critical ρ-value which revealed a correlation between the methods, although not linear according to Pearson’s correlation coefficient. PPV and NPV were calculated with cut-off at 20, 30 and 40 HPC/µL blood where 20 HPC/µL gave an NPV at 100 %. According to the test made, there is correlation between the two methods, but further samples must be analysed to investigate how the results should be compared.

Orientador: Sigmar de Mello Rode / Coorientador: Lucio Murilo dos Santos / Banca: Antonio Olavo Cardoso Jorge / Banca: Fernando Callera / Resumo: Micro-organismos como fungos do gênero Candida, são habitantes comensais da cavidade bucal; em condições normais, co-existem com a microbiota normal sem provocar doenças. Entretanto, alterações locais ou sistêmicas como imunossupressão, desequilíbrio da microbiota oral, hipossalivação e mucosite, secundárias ao tratamento quimioterápico, predispõem pacientes com câncer a um alto risco de infecções fúngicas orais e sistêmicas. O objetivo desse trabalho foi avaliar a presença de leveduras do gênero Candida na cavidade bucal de pacientes onco-hematológicos submetidos a transplante autólogo de células-tronco hematopoiéticas (TACTH). Foram avaliados 27 pacientes, nos períodos pré, e pós-transplante. As amostras da cavidade bucal foram obtidas pela técnica de enxágüe bucal e semeadas em Chromagar Candida para triagem das cepas isoladas. Após o crescimento, foi extraído o DNA e submetido a identificação molecular (PCR) utilizando iniciadores para os genes ribossomais dessa levedura. Após a amplificação do fragmento esperado, as cepas foram sequenciadas utilizando sequenciador automático.Para análise descritiva e estatística dos resultados obtidos, os dados foram submetidos ao teste de Shapiro-Wilk para avaliação da normalidade. Em seguida, o teste de variância Wilcoxon foi utilizado. A significância adotada foi de 5%. Candida spp foi encontrada em 40,74% (11 pacientes), sendo que 2 (18,18%) possuíam mais de uma espécie, dos 9 pacientes colonizados por apenas uma espécie, 7 eram port... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Microorganisms such as fungi of the genus Candida, are commensal inhabitants of the oral cavity; under normal conditions co-exist with normal microflora without causing disease. However, local or systemic changes such as immunosuppression, imbalance in the oral microbiota, hyposalivation and mucositis, secondary to chemotherapy, predispose cancer patients at high risk of oral and systemic fungal infections. The aim of this project is to evaluate the presence of Candida species in the oral cavity of hematological malignances patients undergoing autologous transplantation of hematopoietic stem cells (HSCT). We evaluated 27 patients in preand post-transplant. Samples of the oral cavity were obtained by oral rinse technique and plated on Chromagar Candida for screening of the strains. After growth, the DNA was extracted and subjected to molecular identification (PCR) using primers for the ribosomal genes in this yeast. After amplification of the expected fragment, the strains were sequenced using the sequencer automático. For descriptive and statistical analysis of the results, the data will be submitted to the Shapiro-Wilk test for evaluation of normality. Then the variance Wilcoxon test was used. The significance of 5% was adopted. Candida spp was found in 40.74% (11pacientes), and 2 (18.18%) had more than one species, of the 9 patients colonized by only one species, 7 patients had Candida albicans (77.77%) and 1 Candida dubliniensis (11.11%) and 1 C. krusei (11.11%). Mixed settlements, in the other two patients were composed of C. albicans + C. glabrata; C. albicans + C. dubliniensis. In the second collection (C2), 9 (81.81%) of Candida carriers remained colonized, including by non albicans species. The homology identification of species standardized strains was 85 99%. Within the conditions of this study it was possible to determine with ...(Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Lung disease is the third leading cause of death worldwide. The only curative intervention for end-stage lung disease is lung transplantation, which remains limited by the shortage of viable donor organs. Strategies to improve outcomes for patients with end-stage lung disease include: (i) ex vivo recovery of initially unusable donor lungs to a level suitable for transplantation, and (ii) repair of damaged lungs in situ to avoid the need for transplantation. Recovery of damaged lungs both ex vivo and in situ necessitates precise regulation of the lung extracellular environment, which includes biochemical, physical, and mechanical stimuli across scales. This thesis describes the development of bioengineering tools, including bioreactors and biomaterials, that leverage the lung extracellular environment across cellular, tissue, and organ scales to: (i) recover whole injured donor lungs ex vivo, (ii) assess and repair regional lung tissue injury in situ, and (iii) study the pathological cellular microenvironment in cystic fibrosis. In Chapter 1, regulation of the organ macroenvironment (ventilation, perfusion, systemic metabolism) with a homeostatic cross-circulation bioreactor enabled up to 100 hours of ex vivo lung support and recovery of injured human donor lungs. In Chapter 2, quantitative analysis of localized lung tissue properties, including lung sounds, enabled detection and assessment of pulmonary air leak, and recapitulation of lung microenvironmental features (structure, mechanics, composition) in a therapeutic biomaterial sealant enabled rapid treatment of air leaks. In Chapter 3, the first quantitative characterization of the cystic fibrosis matrisome (matrix proteome) identified pathological alterations to the microenvironment, and investigated implications for inflammation and immunity in cystic fibrosis. Collectively, these studies demonstrate that macro- and microenvironmental signals, including ventilation and perfusion mechanics, homeostatic metabolic regulation, and extracellular matrix structure and composition, can be leveraged to reveal previously unknown drivers of disease and promote recovery and repair of damaged lungs.

South Africa has a low organ donation and transplantation rate despite the availability of medical professionals with the expertise to perform such transplants. This can be attributed to various factors, such as knowledge and attitudes towards organ donation and transplantation. Despite the efforts of the Organ Donor Foundation in South Africa by conducting awareness and education campaigns organ donation rates remains low. There is a wide discrepancy in the rate of organ donation among the different ethnic groups in the country, perhaps due to a lack of knowledge or for cultural or religious reasons. Nurses, as health-care providers, have an important role to play in enabling patients and families to deal with the topic of organ donation. This cross-sectional study investigated the knowledge and attitudes of 268 pre-registration nursing students towards organ donation, at a nursing college in Mthatha, using an anonymous, self- administered questionnaire for data collection. A stratified convenient sampling method was used. The data was captured and analysed using the SPSS statistical package, Version 21; thereafter, descriptive and cross-tabulation analyses were performed on the data. Results: The majority of respondents (62.8%) were aware of organ donation with a small number (1.6%) registered as organ donors. Ethnicity and religion did not influence an individual's decision to donate his/her organs, which suggested that the decision was a personal one. There was no association between age group and willingness to donate a kidney to a relative, although younger respondents were willing to donate kidneys as living donors. There was also no clear relationship between gender and willingness to donate an organ (p-values of 0.03). Knowledge about organ donation was seen as a strong predictor of the attitudes towards organ donation. The majority of respondents were willing to donate organs for transplantation to save the lives of others. It is highly recommended from the results of the study that awareness campaigns to promote organ donation using various strategies and emphasising altruistic motives can increase the organs for donation.

Objective: Judicious selection of potential liver transplant candidates and close monitoring of progress are essential to successful outcomes. Pretransplant psychosocial evaluations are the norm, but the relationship between psychosocial (and neurocognitive status) and longer term medical outcomes is understudied. This exploratory study sought to examine the relationship between objective measures of pretransplant psychosocial and neurocognitive status and service utilization, transplant status, and all-cause mortality. Methods: This retrospective chart review examined outcomes among 108 psychiatric, high-risk liver transplant candidates up to four years following initial evaluation. Predictor variables of outcomes included demographic, medical, neurocognitive, psychological, and mental health treatment variables. Results: Transplant status and neurocognitive functioning were independently associated with all-cause mortality. None of the other variables were associated with outcomes. Conclusions: Better neurocognitive functioning in high-risk liver transplant candidates may allow for greater involvement in medical care and/or compliance with treatment recommendations. More aggressive assessment and management of neurocognitive dysfunction may improve outcomes. Objective measures identified significant psychopathology typical of liver transplant candidates but were not associated with outcomes; engagement in specialized mental health care may have attenuated this relationship. Further study is needed to better understand the relationship between psychosocial functioning and outcomes.

Background: Fanconi anaemia (FA) is a rare genetic disorder associated with bone marrow failure (BMF), congenital anomalies and cancer susceptibility. Stem cell transplantation (SCT) offers a potential cure for BMF or leukaemia, but incurs substantial risks. Little is known about factors influencing SCT decision making. Objective: The study objective was to explore factors influencing patients' with FA and family members' decision making about SCT. Design: Using a mixed-methods exploratory design, we surveyed US and Canadian patients with FA and family members who were offered SCT. Main variables studied: Closed-ended survey items measured respondents' beliefs about the necessity, risks and concerns regarding SCT; multivariable logistic regression was used to examine the association between these factors and the decision to undergo SCT. Open-ended survey items measured respondents' perceptions of factors important to the SCT decision; qualitative analysis was used to identify emergent themes. Results: The decision to undergo SCT was significantly associated with greater perceived necessity (OR = 2.81, P = 0.004) and lower concern about harms of SCT (OR = 0.31, P = 0.03). Qualitative analysis revealed a perceived lack of choice among respondents regarding the use of SCT, which was related to physician influence and respondent concerns about patients' quality of life. Conclusions: Overall, study results emphasize the importance of the delicate interplay between provider recommendation of a medical procedure and patient/parental perceptions and decision making. Findings can help providers understand the need to acknowledge family members' perceptions of SCT decision making and offer a comprehensive discussion of the necessity, risks, benefits and potential outcomes.

Human herpesvirus 6 (HHV-6) infections occur in > 95% of humans. Primary infection, which occurs in early childhood as an asymptomatic illness or manifested clinically as roseola infantum, leads to a state of subclinical viral persistence and latency. Reactivation of latent HHV-6 is common after liver transplantation, possibly induced and facilitated by allograft rejection and immunosuppressive therapy. Since the vast majority of humans harbor the virus in a latent state, HHV-6 infections after liver transplantation are believed to be mostly due to endogenous reactivation or superinfection (reactivation in the transplanted organ). In a minority of cases, however, primary HHV-6 infection may occur when an HHV-6 negative individual receives a liver allograft from an HHV-6 positive donor. The vast majority of documented HHV-6 infections after liver transplantation are asymptomatic. In a minority of cases, HHV-6 has been implicated as a cause of febrile illness with rash and myelosuppression, hepatitis, pneumonitis, and encephalitis after liver transplantation. In addition, HHV-6 has been associated with a variety of indirect effects such as allograft rejection, and increased predisposition and severity of other infections including cytomegalovirus (CMV), hepatitis C virus, and opportunistic fungi. Because of the uncommon nature of the clinical illnesses directly attributed to HHV-6, there is currently no recommended HHV-6-specific approach to prevention. However, ganciclovir and valganciclovir, which are primarily intended for the prevention of CMV disease, are also active against HHV-6 and may prevent its reactivation after transplantation. The treatment of established HHV-6 disease is usually with intravenous ganciclovir, cidofovir, or foscarnet, complemented by reduction in the degree of immunosuppression. This article reviews the current advances in the pathogenesis, clinical diagnosis, and therapeutic modalities against HHV6 in the setting of liver transplantation.

BACKGROUND. Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a risk factor for coronary artery disease (CAD) in nontransplant patients. We evaluated the association between Lp-PLA2, cardiac allograft vasculopathy (CAV) assessed by 3D intravascular ultrasound, and incidence of cardiac adverse events in heart transplant recipients. MATERIALS AND METHODS. Fasting blood samples were obtained and stored from a cross-section of 112 cardiac transplant recipients attending the Mayo cardiac transplant clinic in 2000 to 2001, mean of 4.7 years after transplant. Lp-PLA2 was measured in plasma aliquots using an enzyme-linked immunoassay. Fifty-six of these patients subsequently underwent two 3D intravascular ultrasound studies in 2004 to 2006 12 months apart. Cardiovascular (CV) events included percutaneous coronary intervention, coronary artery bypass grafting (CABG), reduction in left ventricular ejection fraction (LVEF) ≤45% secondary to CAV and CV death. RESULTS. High Lp-PLA2 level was associated with increase in plaque volume (r=0.43, P=0.0026) and percent plaque volume (r=0.45, P=0.0004). The association remained significant after adjusting for clinical and lipid variables. During follow-up of 5.1±1.6 years, 24 CV adverse events occurred in 15 of 112 (13%) heart transplant patients. Lp-PLA2 level>236 ng/mL (higher tertile) identified a subgroup of patients having a 2.4-fold increase of relative risk for combined endpoint of CV events (percutaneous coronary intervention, CABG, LVEF<45%, and CV death; 95% CI 1.16-5.19, P=0.012) compared with patients with Lp-PLA2≤236 ng/mL. CONCLUSIONS. Lp-PLA2 is independently associated with progression of CAV and predicts a higher incidence of CV events and CV death in transplant patients. This finding supports the concept that systemic inflammation is an important mediator of CAV. Lp-PLA2 may be a useful marker for risk of CAV and a therapeutic target in posttransplant patients.

The major obstacle in using pancreatic islet transplantation to cure type I and some type II diabetes is the shortage of the donors. One of ways to overcome such obstacle is to isolate and clone pancreatic stem cells as "seed cells" and induce their differentiation into functional islets as an abundant transplantation source. In this study, a monoclonal human pancreatic stem cell (mhPSC) line was obtained from abortive fetal pancreatic tissues. Pancreatic tissues were taken from abortive fetus by sterile procedures, and digested into single cells and cell clusters with 0.1% type IV collagenase. Cultured in modified glucose-low DMEM with 10% fetal bovine serum (FBS), these single cells and cell clusters adhered to culture dishes, and then primary epidermal-like pancreatic stem cells started to clone. After digesting with 0.25% trypsin and 0.04% EDTA, fibroblasts and other cells were gradually eliminated and epithelioid pancreatic stem cells were gradually purified during generations. Using clone-ring selection, the mhPSCs were obtained. After addition of 10 ng/mL epidermal growth factor (EGF) in cell culture medium, the mhPSCs quickly grew and formed a gravelstone-like monolayer. Continuously proliferated, a mhPSC line, which was derived from a male abortive fetus of 4 months old, has been passed through 50 generations. More than 1×10 9 mhPSCs were cryo-preserved in liquid nitrogen. Karyotype analysis showed that the chromosome set of the mhPSC line was normal diploid. Immunocytochemistry results demonstrated that the mhPSC line was positive for the pdx1, glucagon, nestin and CK19, and negative for the insulin, CD34, CD44 and CD45 protein expression. RT-PCR revealed further that the mhPSCs expressed transcription factors of the pdx1, glucagon, nestin and CK19. Also, in vitro induced with β-mercaptoethanol, the mhPSCs differentiated into nerve cells that expressed the NF protein. Induced with nicotinamide, the mhPSCs differentiated into functional islet-like clusters, as identified by dithizone staining, which expressed the transcription factor of the insulin and secreted the insulin and C-peptide. Furthermore, the transplantation of mhPSCs-induced pancreatic islets into the subcapsular region of the kidney in streptozotocin-induced diabetic rats could reduce blood glucose levels and prolong the life time.