Purpose: The purpose of this study was to determine the usefulness of SBAR-cued web-based communication skills training and address study participants' perceptions of the training. Specific Aims: Evaluate the usefulness and accuracy of participants to answer prompts of SBAR-cued communication responses. Describe individuals' perspectives of the acceptability of using SBAR patient-HCP communication simulation to better prepare for ED visits during a SCC. Framework: This study was guided by The Theory of Self-Care Management for Sickle Cell Disease (SCMSCD). Design: A secondary analysis was conducted using a qualitative descriptive approach. Inter-rater reliability (IRR) of qualitative data was used to evaluate the usefulness and accuracy of participants to answer prompts of SBAR-cued communication responses. Content analysis was also utilized to describe individuals' perspectives of the acceptability of using SBAR patient-HCP communication simulation to better prepare for ED visits during a SCC. Results: IRR between raters ranged from 64%-94% with predominant themes of (1) Patient-Provider Communication and Interaction, (2) Patients want to be Heard and Believed, (3) Accuracy of the ED Experience and Incorporating the Uniqueness of each Patient and (4) Overall Usefulness of the Video Trainer emerging. Conclusions: This secondary analysis supported how SBAR can be effectively used to assist patients in a SCC to communicate with their HCP. Participants' responses indicated the training module facilitated communication between patients and HCPs.

Endemic Burkitt lymphoma (eBL) is the most common pediatric cancer in malaria-endemic equatorial Africa and nearly always contains Epstein-Barr virus (EBV), unlike sporadic Burkitt Lymphoma (sBL) that occurs with a lower incidence in developed countries. Despite this increased burden the study of eBL has lagged. Additionally, while EBV was isolated from an African Burkitt lymphoma tumor 50 years ago, however, the impact of viral variation in oncogenesis is just beginning to be fully explored. In my thesis research, I focused on investigating molecular genetics of the endemic form of this lymphoma with a particular emphasis on the role of the virus and its variation in pathogenesis using novel sequencing and bioinformatic strategies. First, we sought to understand pathogenesis by investigating transcriptomes using RNA sequencing (RNAseq) from 30 primary eBL tumors and compared to sBL tumors. BL tumor samples were prospectively obtained from 2009 until 2012 in Kenya. Within eBL tumors, minimal expression differences were found based on anatomical presentation site, in-hospital survival rates, and EBV genome type; suggesting that eBL tumors are homogeneous without marked subtypes. The outstanding difference detected using surrogate variable analysis was the significantly decreased expression of key genes in the immunoproteasome complex in eBL tumors carrying type 2 EBV compared to type 1 EBV. Secondly, in comparison to previously published pediatric sBL specimens, the majority of the expression and pathway differences were related to the PTEN/PI3K/mTOR signaling pathway and was correlated most strongly with EBV status rather than the geographic designation. Moreover, the common mutations were observed significantly less frequently in eBL tumors harboring EBV type 1, with mutation frequencies similar between tumors with EBV type 2 and without EBV. In addition to the previously reported genes, we identified a set of new genes mutated in BL. Overall, these suggested that EBV, particularly EBV type 1, supports BL oncogenesis alleviating the need for particular driver mutations in the human genome. Second, we sought to comprehensively define sequence variations of EBV across the viral genome in eBL tumor cells and normal infections, and correlate variations with clinical phenotypes and disease risk. We investigated the whole genome sequence of EBV from primary tumors (N=41) and plasma from eBL patients (N=21) as well as EBV in the blood of healthy children (N=29) within the same malaria endemic region. We conducted a genome wide association analysis study with viral genomes of healthy kids and BL kids. Furthermore, we found that the frequencies of EBV types among healthy kids were at equal levels while they were skewed in favor of type 1 (70%) among eBL kids. To pinpoint the fundamental divergence between viral genome subtypes, type 1 and type 2, we constructed phylogenetic trees comparing to all public EBV genomes. The pattern of variation defined the substructures correlated with the subtypes. This investigation not only deciphers the puzzling pathogenic differences between subtypes but also helps to understand how these two EBV types persist in the population at the same time. Overall, this research provides insight into the molecular underpinning of eBL and the role of EBV. It further provides the groundwork and means to unravel the complexity of EBV population structure and provide insight into the viral variation that may influence oncogenesis and outcomes in eBL and other EBV-associated diseases. In addition, genomic and mutational analyses of Burkitt lymphoma tumors identify key differences based on viral content and clinical outcomes suggesting new avenues for the development of prognostic molecular biomarkers and therapeutic interventions.

The CBFbeta-SMMHC fusion protein is expressed in acute myeloid leukemia (AML) samples with the chromosome inversion inv(16)(p13;q22). This fusion protein binds the transcription factor RUNX with higher affinity than its physiological partner CBFbeta and disrupts the core binding factor (CBF) activity in hematopoietic stem and progenitor cells. Studies in the Castilla laboratory have shown that CBFbeta-SMMHC expression blocks differentiation of hematopoietic progenitors, creating a pre-leukemic progenitor that progresses to AML in cooperation with other mutations. However, the combined function of cumulative cooperating mutations in the pre-leukemic progenitor cells that enhance their expansion to induce leukemia is not known. The standard treatment for inv(16) AML is based on the use of non-selective cytotoxic chemotherapy, resulting in a good initial response, but with limited long-term survival. Therefore, there is a need for developing targeted therapies with improved efficacy in leukemic cells and minimal toxicity for normal cells. Here, we used conditional Nras+/LSL-G12D; Cbfb+/56M; Mx1Cre knock-in mice to show that allelic expression of oncogenic N-RasG12D expanded the multi-potential progenitor (MPP) compartment by 8 fold. Allelic expression of Cbfbeta-SMMHC increased the MPPs and short-term hematopoietic stem cells (ST-HSCs) by 2 to 4 fold both alone and in combination with N-RasG12D expression. In addition, allelic expression of oncogenic N-RasG12D and Cbfbeta-SMMHC increases survival of pre-leukemic stem and progenitor cells. Differential analysis of bone marrow cells determined that Cbfb+/MYH11 and Nras+/G12D; vii Cbfb+/MYH11 cells included increased number of blasts, myeloblasts and promyelocytes and a reduction in immature granulocytes, suggesting that expression of N-RasG12D cannot bypass Cbfbeta-SMMHC driven differentiation block. N-RasG12D and Cbfbeta-SMMHC synergized in leukemia, in which Nras+/G12D; Cbfb+/MYH11 mice have a shorter median latency than Cbfb+/MYH11 mice. In addition, the synergy in leukemogenesis was cell autonomous. Notably, leukemic cells expressing N-RasG12D and Cbfbeta-SMMHC showed higher (over 100 fold) leukemia-initiating cell activity in vivo than leukemic cells expressing Cbfbeta-SMMHC (L-IC activity of 1/4,000 and 1/528,334, respectively). Short term culture and biochemical assays revealed that pre-leukemic and leukemic cells expressing N-RasG12D and Cbfbeta-SMMHC have reduced levels of pro-apoptotic protein Bim compared to control. The Nras+/G12D; CbfbMYH11 pre-leukemic and leukemic cells were sensitive to pharmacologic inhibition of MEK/ERK signaling pathway with increasing apoptosis and Bim protein levels but not sensitive to PI3K inhibitors. In addition, knock-down of Bcl2l11 (Bim) expression in Cbfbeta-SMMHC pre-leukemic progenitors decreased their apoptosis levels. In collaboration with Dr. John Bushweller’s and other research laboratories, we recently developed a CBFbeta-SMMHC inhibitor named AI-10-49, which specifically binds to CBFbeta-SMMHC, prevents its binding to RUNX proteins and restores CBF function. Biochemical analysis in human leukemic cells showed that AI-10-49 has significant specificity in reducing the viability of leukemic cells expressing CBFbeta-SMMHC (IC50= 0.83μM), and negligible toxicity in normal cells. Likewise, mouse Nras+/G12D; viii Cbfb+/MYH11 leukemic cells were sensitive to AI-10-49 (IC50= 0.93μM). By using the NrasLSL-G12D; Cbfb56M mouse model, we also show that AI-10-49 significantly prolongs the survival of mice bearing the leukemic cells. Preliminary mechanistic analysis of AI-10-49 activity has shown that AI-10-49 increased BCL2L11 transcript levels in a dose and time dependent manner in murine and human leukemic cells, suggesting that the viability through BIM-mediated apoptosis may be targeted by both oncogenic signals. My thesis study demonstrates that Cbfbeta-SMMHC and N-RasG12D promote the survival of pre-leukemic myeloid progenitors primed for leukemia by activation of the MEK/ERK/Bim axis, and define NrasLSL-G12D; Cbfb56M mice as a valuable genetic model for the study of inv(16) AML targeted therapies. For instance, the novel CBFbeta-SMMHC inhibitor AI-10-49 shows a significant efficacy in this mouse model. This small molecule will serve as a promising first generation drug for targeted therapy of inv(16) leukemia and also a very useful tool to understand mechanisms of leukemogenesis driving by CBFbeta-SMMHC.

No description available.

Hematopoietic stem cells (HSCs) have historically been difficult to maintain ex vivo with many attempts to culture them in vitro by mimicking their natural biological environment. Providing a hypoxic environment is one way to achieve this goal and can be performed by using hypoxia stimulating compounds that inhibits the degradation of HIF1a which plays an important role in regulating hypoxia. For each sample 50 murine HSCs were isolated with fluorescence-activated cell sorting (FACS) and cultured with different concentrations of the hypoxia inducible compound Adaptaquin for 13 days followed by analysing with flow cytometry. The results showed an increase in proliferation of treated cells with the highest average total viable cell count for cells treated with 100 nM Adaptaquin of 4,70 ± 1,12 x 105 cells compared to the control which had 2,39 ± 0,76 x 105 cells. The HSC frequency was highest in the control samples with an average of 1,91 ± 0,42 % compared to the 5 mM treated samples with the highest average HSC frequency which was 1,52 ± 0,82 %. The biggest noticeable difference between the control and treated samples was seen when observing the total cell count. The difference in proliferation was on the other hand too small to see significant difference between the samples. The conclusion is that Adaptaquin did not have any significant impact on keeping the cells undifferentiated but could have a potential to be used as a compliment to other factors to maintain HSCs in vitro and to mimic its hypoxic biological environment. / Hematopoetiska stamceller (HSCs) har historiskt sett varit svåra att odla ex vivo och många försök har genomförts in vitro genom att efterlikna deras naturliga biologiska miljö. Att tillhandahålla en hypoxisk miljö är en metod för att uppnå detta och kan göras med användning hypoxi-stimulerande substanser som hämmar nedbrytningen av HIF1a som spelar en viktig roll i regleringen av hypoxi. För varje prov isolerades 50 murina HSCs med fluorescence-activated cell sorting (FACS) och odlades med olika koncentrationer av det hypoxi-inducerande ämnet Adaptaquin under 13 dagar följt av analys med flödescytometri. Resultaten visade en ökning i avseende på proliferationen hos behandlade celler där det högsta genomsnittliga totala antalet levande celler behandlade med 100 nM Adaptaquin som var 4,70 ± 1,12 x 105 celler jämfört med kontrollen som hade 2,39 ± 0,76 x 105 celler. HSC-frekvensen var högst i kontrollproverna med ett genomsnitt på 1,91 ± 0,42 % jämfört med proverna behandlade med 5 mM Adaptaquin som hade den högsta genomsnittliga HSC-frekvensen som låg på 1,52 ± 0,82 %. Den största synliga skillnaden mellan kontroll- och behandlingsprover var synlig när det observerade totala antalet celler jämfördes mellan behandlade prover som i genomsnitt hade fler totala celler. Skillnaden i proliferation var å andra sidan för liten för att se en signifikant skillnad mellan proverna. Slutsatsen är att Adaptaquin inte hade någon signifikant påverkan på att hålla HSCs odifferentierade men kan ha potential att användas som ett komplement till andra faktorer för att odla HSCs in vitro och efterlikna dess hypoxiska biologiska miljö.

Allt mer forskning kring digitala bildanalysplattformar sker då de har visat lika bra eller bättre prestation än existerande manuella och automatiserade metoder samt effektiviserat handläggningstiden för patientprov. Nyligen har företaget CellaVision utvecklat ett prototypprogram som via bildanalys kan räkna och klassificera retikulocyter. Syftet med studien var att jämföra noggrannheten och precisionen av CellaVisions prototypprogram för retikulocytnumrering med manuell mikroskopering och en automatiserad cellräknare. Totalt 40 blodutstryk tillverkades av 20 blodprov med abnormala värden retikulocyter (>2,5%) och 20 med normala värden retikulocyter (0,5-2,5%). Blodutstryken analyserades med prototypprogrammet, manuell mikroskopering och den automatiserade cellräknaren varpå retikulocytkoncentrationen för respektive resultat jämfördes med Pearsons korrelationstest och Bland-Altmananalys. Blodutstryk tillverkades av tre nivåer retikulocytkontroller och analyserades flertal gånger med prototypprogrammet vars retikulocytkoncentration jämfördes med andra metoders retikulocytkoncentrationer. Samma blodutstryk analyserades med olika antal inskannade celler för att utvärdera hur variationen påverkades. Resultaten från komparabilitetsstudien visade att prototypprogrammet erhöll en utmärkt korrelation och en god överensstämmelse med de andra metoderna. Precisionsstudien visade att prototypprogrammet erhöll konsekventa retikulocytkoncentrationer vid repeterade analyser och en låg variationskoefficient (0,25-10%) vid högre antal inskannade celler vid tre olika nivåer av retikulocytkontroller. Resultaten visar CellaVisions prototypprogram designat för retikulocyträkning erhåller en god noggrannhet och en hög precision vid jämförelse med manuell mikroskopering och en automatiserad cellräknare, dock behöver större studier utföras då antalet patientprov var begränsade. / More and more research on digital image analysis platforms is taking place as they have shown to have as good or better performance than existing manual and automated methods and have also streamlined the processing time for patient samples. Recently, the company CellaVision has developed a prototype program that can enumerate and classify reticulocytes via image analysis. The purpose of the study was to compare the accuracy and precision of CellaVision's prototype program for reticulocyte numbering with manual microscopy and an automated cell counter. A total of 40 blood smears were made from 20 blood samples with abnormal reticulocyte values (>2,5%) and 20 with normal reticulocyte values (0,5-2,5%). The blood smears were analysed with the prototype program, manual microscopy, and the automated cell counter, after which the reticulocyte concentration for each result was compared with Pearson's correlation test and Bland-Altman analysis. Blood smears were made by three levels of reticulocyte controls and analysed several times with the prototype program whose reticulocyte concentration was compared with the other methods' reticulocyte concentrations. The same blood smears were analysed with different numbers of scanned cells to evaluate how the variation was affected. The results of the comparability study showed that the prototype program obtained an excellent correlation and a good agreement with the other methods. The precision study showed that the prototype program obtained consistent reticulocyte concentrations in repeated assays and a low coefficient of variation (0,25-10%) in higher numbers of scanned cells at three different levels of reticulocyte controls. The results showed the CellaVision's prototype program designed for reticulocyte counting obtained good accuracy and high precision when compared with manual microscopy and an automated cell counter, however, larger studies need to be performed as the number of patient samples was limited.

Discussion of the hematologic complications of vaccination for severe acute respiratory syndrome coronavirus-2 (COVID-19) has primarily focused on the development of vaccine-associated immune thrombosis with thrombocytopenia (VITT). Other hematologic complications are uncommon. We report the case of a patient who developed immunoglobulin G (IgG)-mediated autoimmune hemolytic anemia (AIHA) after the Moderna COVID-19 messenger ribonucleic acid (mRNA) vaccine.

No description available.

Hematology analyzers can be used for screening patients for blood abnor-malities. The techniques used in a hematology analyzer include impedanceanalysis, flow cytometry and spectroscopy, which allow for measuring of forexample absolute count, sizes and concentration of different cells in a patient’sblood sample. Hyperlipidemia, which refers to elevated blood lipid levels, isthe primary cause of heart-related illness and fatalities in today’s developedor developing countries. Currently, blood lipid levels are not measured as aparameter with hematology analyzers. Since hematology analyzers allow for arapid general screening of blood parameters, an area of interest is therefore tobe able to measure blood lipids with a hematology analyzer. Thus, this studyaims to investigate the possibility of detecting and measuring blood lipids witha hematology analyzer, using flow cytometry and/or spectrophotometry. In order to investigate this possibility, two simulating methods were conductedwhere in the first method Intralipid 20% was mixed with saline into sampleswith different lipid concentrations. In the second method, diluent wasused instead of saline. Lastly a Correlation study was performed whereIntralipid 20% was mixed with donor blood to prepare samples with differentlipid concentrations. All samples were then analyzed in a hematologyanalyser and scatter plots from flow cytometry and light absorption datafrom spectrophotometry measurements were obtained. The methods showedthat there is a strong correlation between number of detected pulse countsfrom the scatter plots and lipid concentration. Same applies to lightabsorption compared to the lipid concentration of the samples, measured withspectrophotometry. The results from this study show that it is in fact possible to detect andmeasure blood lipid levels with a hematology analyser using flow cytometryand spectrophotometry. Further development within this area could thereforeenable simple screening of this additional parameter and early detection ofindications of hyperlipidemia. / Hematologianalysatorer möjliggör screening av eventuella avvikelser ipatienters blood. De tekniker som används i en hematologianalysatorinkluderar impedansanalys, flödescytometri och spektroskopi, vilka möj-liggör mätning av till exempel absolutantal, storlekar och koncentrationav olika celler i ett patientblodprov. Hyperlipidemi, vilket hänvisar tillförhöjda blodlipidnivåer, är den främsta orsaken till hjärtrelaterade sjukdomaroch dödsfall i dagens utvecklade eller utvecklingsländer. För närvarandemäts inte blodlipidnivåer som en parameter med hematologianalysatorer.Eftersom hematologianalysatorer möjliggör en snabb allmän screening avblodparametrar, är ett intresseområde därför att kunna mäta blodlipider meden hematologianalysator. Syftet med denna studie är därför att undersökamöjligheten att detektera och mäta blodlipider med en hematologianalysator,med hjälp av flödescytometri och/eller spektrofotometri. Två simuleringsmetoder genomfördes, där den första metoden innefattadeblandning av Intralipid 20% med saltlösning till prover med varierandelipidkoncentration. I den andra metoden användes spädningsvätska iställetför saltlösning. Slutligen genomfördes en korrelationsstudie där Intralipid20% blandades med donatorblod och prover med olika lipidkoncentrationerförbereddes. Alla prover analyserades sedan i en hematologianalysator ochspridningsdiagram och ljusabsorptionsdata från spektrofotometrimätningarerhölls. Resultaten visade att det finns en stark korrelation mellan antalet de-tekterade pulsräkningar från spridningsdiagrammen och lipidkoncentrationen.Samma gäller för ljusabsorption jämfört med lipidkoncentrationen i proverna,mätt med spektrofotometri. Resultaten från denna studie visar att det faktiskt är möjligt att detektera ochmäta blodlipidnivåer med en hematologianalysator med hjälp av flödescyto-metri och spektrofotometri. Vidare utveckling inom detta område skulle därförkunna möjliggöra enkel screening av patienters blod lipidkoncentration samtunderlätta en tidig upptäckt av indikationer på hyperlipidemi.

A Ararajuba (Guaruba guarouba) é um psitacídeo de médio porte, endêmico do Bioma Amazônico, que se encontra em perigo de extinção devido, principalmente, à perda de habitat e ao tráfico de animais. Poucos trabalhos científicos foram desenvolvidos para avaliar as condições sanitárias dessas aves mantidas em cativeiro, visando à reintrodução da espécie na natureza. Este estudo teve por objetivo avaliar o estado sanitário das aves mantidas em Zoológicos e Criadouros do Estado de São Paulo, por meio de avaliações hematológica, microbiológica e parasitológica. Foram colhidas amostras de 87 indivíduos mantidos em seis Parques Zoológicos, três Criadouros Comerciais e um Criadouro Conservacionista, obtendo-se como resultados da avaliação hematológica: valor médio do volume globular (Ht%): 46,12; valor médio de proteína total sérica (g/dL): 3,55; valor médio de hemoglobina (g/dL): 12,68; valor médio do número de hemácias (x 106 mm3): 3,52; valor médio do número dos leucócitos totais (x103/mm3): 13,41; valor médio do número de trombócitos (/mm3): 26.029,4; valor médio do VCM (fL): 133,3; valor médio do HCM (pg): 36,58; valor médio do CHCM (g/L) : 27,65; valor médio de heterófilos (%):56.62; valor médio de linfócitos (%): 42,22; valor médio de monócitos (%): 0,9; valor médio de eosinófilos (%):0,04; valor médio de basófilos (%): 0,20. Quanto à análise microbiológica, não houve isolamento de Salmonella spp. na microbiota intestinal, porém, houve crescimento de Escherichia coli em 50% dos animais. No exame parasitológico, todos os animais foram negativos tanto para a pesquisa de endoparasitas intestinais como para hemoparasitas. O soro obtido da centrifugação do sangue e a papa de hemácias foram armazenados para futuras pesquisas. / The Ararajuba or Golden conure (Guaruba guarouba) is a medium-sized parrot, endemic to the Amazon biome and threatened of extinction primarily due to habitat loss and poaching. Few scientific studies have examined the health conditions of these birds kept in captivity, which will be essential to future reintroduction efforts. This study aimed to evaluate the health status of Ararajubas maintained in zoos and private breeders in São Paulo state, Brazil, through haematological, microbiological and parasitological evaluations. Samples were obtained from 87 individuals held at six Parks Zoos, three Commercial Breeders and one Conservation Breeder. Average haematological results: packed cell volume 46.12%, total serum protein 3.55 g/dL, haemoglobin 12.68 g/dL, erythrocytes 3.52 x 106 cells/mm3, Leukocytes 13.41 x 103 cells/mm3, thrombocytes 26.03 x 103 cells/mm3, Mean Corpuscular Volume 133.3 fL, Mean Corpuscular Haemoglobin 36.58 pg, Mean Corpuscular Haemoglobin Concentration 27.65 g/dL, heterophils 56.62%, lymphocytes 42.22%, monocytes 0.9%, eosinophils 0.04%, basophils 0.20%. Microbiological cultures: Escherichia coli could be isolated from 50% of the cloacal swabs and Salmonella spp. was not isolated in none. Parasitology: all animals were negative for both intestinal parasites and haemoparasites. Plasma and erythrocytes obtained by centrifugation were stored for future research.