Background: Cancer is a common disease, and the choice of treatment becomes more difficult over time due to chemotherapy resistant in cancer cells. To improve the in vitroassay and the individual cancer treatment, a luminescence-based endpoint assay, CellTiter Glo 2.0 was compared with the currently in use fluorescence endpoint assay, fluorometric microculture cytotoxic assay.  Aim: The aim of this study was to validate and compare the CellTiter Glo 2.0 assay with awell-established method (FMCA) and MTT [3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2Htetrazolium bromide] assay. Moreover, investigate whether the generated data can be used as a reference database for validation of patient samples in the future.   Materials and methods: The validation was performed on peripheral blood mononuclear cells from different healthy donors and two cell lines (HCT116-wt and HT-29) of colorectal cancer carcinoma were ordered frozen from American Type Culture Collection. Analysis was also done in solid samples (ovarian and kidney cancer cells). To get as correct evaluation as possible all materials were analyzed in parallel between the two methods.  Results and conclusion: A clear trend was observed when using CellTiter Glo 2.0 assay,post FMCA directly on tumor cells. This setup, makes it possible to collect reference data in the future. In addition, a high spread of the survival index data was noted between the two methods. The reason is still unknown but could be due to the low number of tested tumor cells, therefore more tumor cells need to be tested in future studies

I denna metodstudie jämfördes Thermo Fishers bevarande lösningar RNAlater och RNAlater-ICE med frystorkning som bevaring av skelettmuskulatur från människa. Studien gjordes med syftet att avgöra om RNAlater och/eller RNAlater-ICE kan ersätta frystorkning, i hopp om att underlätta dissekering av muskelvävnad. De molekylärbiologiska analyser som utfördes var proteinbestämning; Western Blot inriktad på proteinerna mTor, S6, S6K1, eEF2, myosin typ II och aktin; samt mätning av glykogen och citratsyntasaktivitet. Därtill påbörjades även en aminosyraanalys. Studien utfördes hos William Apró på Åstrandlaboratoriet på Gymnastik- och idrottshögskolan i Stockholm. Resultaten visade att både RNAlater och RNAlater-ICE är olämpliga att använda i studier som innefattar samtliga av de genomförda analyserna. Detta då analysresultaten från de alternativa bevaringsmetoderna och de från frystorkningen inte var likvärdiga. Därmed drogs slutsatsen att varken RNAlater eller RNAlater-ICE kan ersätta frystorkning som bevaringsmetod i Aprós vidare studier. / In this method study Thermo Fisher’s preserving solutions RNAlater and RNAlater-ICE were compared to lyophilization for preservation of human skeletal muscle. The study was conducted with the aim of determining whether RNAlater and / or RNAlater-ICE can replace lyophilization, in the hope of facilitating dissection of muscle tissue. The molecular biological analyzes performed were protein assay; Western Blot focused on the proteins mTor, S6, S6K1, eEF2, myosin type II and actin; alongside of measurments on glycogen and citrate synthase activity. In addition, an amino acid analysis was initiated. The study was executed with William Apró at the Åstrand Laboratory at The Swedish School of Sport and Health Science in Stockholm. The results showed that both RNAlater and RNAlater-ICE are unsuitable for use in studies that include all of the analyzes performed. This is because the analysis results from the alternative preservation methods and those from lyophilization were not equivalent. Thus, it was concluded that neither RNAlater nor RNAlater-ICE can replace lyophilization as a preservation method in Apró’s further studies.

<p>  </p> <p>In order to comprehend complex and abstract phenomena, students must partake in the process of learning by integrating complex and invisible components without ever physically encountering or manipulating such components. Prior knowledge and experiences will influence the way students assimilate and model new experiences and knowledge. If prior knowledge possesses a degree of non-normative conceptions, students' understanding of abstract phenomena may diverge dramatically from accepted scientific explanations. Embodied cognition proposes that learning about natural phenomena can develop from information gained via interactions between the body and the physical environment. Multimodal experiences can shape students' conceptual understanding of abstract phenomena.</p> <p>Incorporating technology tools to explore science concepts is a trend utilized to give high-quality education. The use of physical and virtual manipulation tools in science instruction has favored the improvement of modeling science phenomena in general. Visuohaptic simulations are also learning manipulatives that blend physical and virtual manipulation affordances as a unison experience. </p> <p>The current dissertation proposed the implementation of a learning experience where students engage in experimentation with a visuo-haptic simulation to explore and model reaction forces on truss structures. The study examined undergraduate students’ conceptual understanding, graphical representations, and the modeling refinement process of reaction forces on truss structures before, during, and after engaging with visuo-haptic simulation on truss structures using different modalities. A design-based research methodology was implemented to design, explore, and refine a learning experience with a visuo-haptic simulation of truss structures through two research phases. The learning experience occurred as a laboratory activity in a statics course at a Midwest university.</p> <p>The first phase of this dissertation investigated students' conceptual understanding and graphical representations of reactive forces on a complex truss structure by interacting with a visuo-haptic simulation of truss structures. Students participated in two treatment groups: visuo-haptic exposure and visual-only exposure. The results of the first phase suggested that students that engaged with the visuo-haptic simulation using different modalities improved their conceptual understanding of truss structures significantly. Moreover, students exposed to haptic feedback significantly improved their graphical representations on tasks where the haptic feedback was involved. </p> <p>The second phase of the current dissertation examined students’ developing models of reactive forces on a truss structure before, during, and after engaging with a visuo-haptic simulation of truss structures. Students participated in two sequential treatment groups: visual to visuo-haptic and haptic to visuo-haptic. The quantitative results suggest that both treatment groups performed significantly better in their model representations after being exposed to the learning experience but show no difference across treatment groups. The qualitative results suggest that the visual to visuo-haptic group interpreted their experiences much more coherently, leading to a more sophisticated version of their model of reaction forces on truss structures. </p>

Ultraljudsundersökning av arteria carotis interna (ICA) och arteria carotis externa (ECA) utförs på fysiologiska kliniker för att bland annat bedöma blodflödeshastigheten i kärlen. Det kan i sin tur användas vid diagnostik av plack och stenoser. Behandling av stenos är viktigt eftersom det kan leda till stroke och transitorisk ischemisk attack (TIA). Syftet med den här studien är att jämföra den maximala blodflödeshastigheten i ECA och ICA, samt om den skiljer sig mellan höger och vänster sida. I studien inkluderades 36 deltagare i åldern 20–27 år. Resultatet analyserades med ett parat t-test samt med en korrelationsanalys. En statistisk signifikant skillnad identifierades vid jämförelse av ICA och ECA på samma sida. Ingen signifikant skillnad observerades vid sidojämförelse av vänster och höger ECA respektive ICA. En korrelation påvisades mellan vänster och höger ECA samt mellan vänster och höger ICA. Ytterligare en signifikant korrelation identifierades mellan ECA bilateralt och ICA dx. Slutsatsen med studien är att det finns en statistisk skillnad mellan flödeshastigheten i ICA och ECA och därav går det inte att använda samma referensvärden till båda kärlen. / Ultrasound examination of the internal carotid artery (ICA) and the external carotid artery (ECA) is performed in physiological clinics to assess the blood flow velocity in the vessels. Furthermore, it can be used in the diagnosis of plaque and stenoses. Treatment of stenosis is important as it can lead to stroke and transient ischemic attack (TIA). The purpose of this study is to compare the maximum blood flow velocity in ECA and ICA and allocate a possible difference between the right and left side. The study included 36 participants between the ages of 20–27. The result was analyzed using a paired t-test and a correlation analysis. A statistical significance was identified when comparing ICA and ECA on the same side. No significant difference was found when comparing left and right ECA and ICA. One correlation was detected between left and right ECA and between left and right ICA. Another correlation was identified between ECA bilaterally and ICA dexter. The conclusion of this study is that there is a significant difference between the flow velocity in ICA and ECA and therefore it is not possible to use the same reference values ​​for both vessels.

Akut hjärtinfarkt är globalt sett en vanlig dödsorsak. Hjärtinfarkter behöver inte ge tydliga symptom och dödsorsaken kan därmed vara okänd fram till en klinisk obduktion och att histopatolgiska studier av hjärtmuskelvävnad genomförs. Fosforwolframsyra-hematoxylin (PTAH) är en färgningsmetod som kan användas vid visualisering av hjärtinfarkt genom att färga myokardceller, fibrin, kontraktionsband och cellkärnor blå medan kollagen färgas rosa till rödbrunt. Syftet med examensarbetet var att optimera PTAH-färgningsprotokoll för att underlätta diagnosticering av hjärtinfarkt. Tre olika färgningsprotokoll, två med PTAH med Mallory Bleach (#1 och #2) och ett med PTAH med refixering i Bouins lösning (#3), jämfördes vid inkubering över natt i rumstemperatur samt 3-4 h i värmeskåp vid 56 ºC. Vävnader som färgades in var från paraffininbäddade klossar vilka tillhörde utsvarade fall med konstaterad hjärtinfarkt. Dessa snittades och värmdes fast på oladdade samt laddade objektglas. Efter färgning graderades infärgning efter kategorierna vävnadsdifferentiering och cellkomponenter med en poängskala från 0, ej bedömbar, till 3, optimal. Det protokoll med högst poäng optimerades för att hitta bästa inkuberingstid i PTAH-lösning, därefter implementerades protokollet genom att bekräfta korrekt infärgning vid upprepade infärgningar av hjärtmuskelvävnad. Protokoll #3 visade på högst poäng efter infärgning vid jämförelsen med protokoll #1 och #2 och under optimering framgick inkubering under 4 h i värmeskåp vid 56 ºC som mest gynnsam, då denna inkuberingstid även visade på mindre känslighet vid dehydrering. Slutsatsen blev att PTAH-färgning efter protokoll #3 med refixering i Bouins lösning med 4 h i värmeskåp gav bäst resultat, men färgnyansen varierar med tid mellan dödsfallet och obduktionen. / Acute myocardial infarction (MI) is a common cause of death globally. MI without symptoms or with atypical symptoms is usually first detected at clinical autopsy with histological analyses of the myocardium. Phosphtungstic acid haematoxylin (PTAH) is a method to visualize MI by staining myocytes, fibrin, contraction bands and nuclei blue while collagen stains reddish brown. The aim of this degree project is to optimize PTAH staining protocol for MI diagnosis. Three different staining protocols, two protocols with PTAH staining and Mallory Bleach solution and one protocol with PTAH staining with refixation in Bouin’s solution (#1-3), where compared along with incubation overnight in room temperature and at different hours in heat (56 ºC). Paraffin embedded tissues from myocardium from different autopsy cases diagnosed with MI were cut before mounting and heating on non-charged as well as charged slides. After staining, results were evaluated using scores 0-3 for two parameters; differences in tissues and cell components. Highest evaluated protocol where optimized to find the ultimate incubation in PTAH staining solution. For implementations, additional myocardial tissues from other cases were stained to confirm repeated results. Protocol #3 was evaluated higher in comparison with #1-2 and optimized to 4 h incubation in PTAH-solution in heating with staining results less sensitive to dehydration. In conclusion, staining protocol #3 with 4 h heating was optimal, however, vulnerable to prolonged morgue storage time losing the insensitivity of colour.

<p> Universities have increased the number of fully or partially online courses they offer to meet students' family, work-life, and academic needs. As a result of this shift, the HyFlex learning paradigm, which provides hybrid and flexible learning options, has gained traction in academia in recent years. The COVID-19 pandemic has altered the standards for academic levels of success in higher education for the foreseeable future. In order to offer a sense of belonging and sustain an equal or greater degree of engagement, instructors have been compelled to try and replicate the rich interaction of face-to-face situations in a virtual context. During the pandemic, the results of studies and surveys revealed a story of student dissatisfaction due to a lack of involvement, participation, sense of community, lack of faculty readiness, and technology concerns. Evidence from the past has supported the fact that synchronous online modality not only facilitates the tasks of a traditional face-to-face learning environment, but also has some distinct advantages over conventional approaches. Students' commitment to stay connected in the learning experience is linked to their sense of belonging to a learning cohort. It is vital to create a learning environment in which students feel a part of a learning community and actively participate in the learning process in order to foster knowledge generation. An Interactive Synchronous HyFlex model, intended to help students feel committed and engaged in their learning community through these uncertain times and beyond, is being explored as part of this research. The approach is studied using a Community of Inquiry (CoI) framework, as communities are a crucial part of effective student-centered learning settings. A convergent parallel mixed methods case study design was used to gain a deeper knowledge of the usefulness of the new HyFlex model under study. Students enrolled in the Interactive Synchronous HyFlex design thinking course during the Fall 2020 semester were the study's participants. The quantitative phase of the study looked into a) the students’ perception of overall teaching, social and cognitive presences in the HyFlex design thinking classroom; and b) if there was any change in students’ perception of community of inquiry based on their mode of participation (face-to-face/remote). The qualitative part of the study looked in-depth at the lived experiences of students in the HyFlex design thinking classrooms throughout the semester. The integration and interpretation of the two phases provides a positive student perspective of the Interactive Synchronous HyFlex model, and it helps to observe how the community of inquiry has played out in HyFlex design thinking classrooms. </p>

Heart failure with preserved ejection fraction has its main pathophysiology in the diastolic dysfunction which is characterized by increased filling pressures. This can be discovered during a routinely performed echocardiographic examination. In 2016 the American Society of Echocardiography published a recommendation to simplify the assessment of diastolic dysfunction. Whether or not these recommendations have simplified the gradings is questionable. Left atrial strain has been shown to decrease in the absent of diastolic dysfunction and 3D echocardiographic volume measurements of the left atrium has been showing more accurate volumes of the heart compared to 2D-measurements. A quantitative study was therefore made to compare the results from the recommended algorithm with the two methods - left atrial strain and 3D-measurements of the left atrium volume. A total of 138 women and men were included with the data collected both prospective and retrospective. The aim with this quantitative study was to examine if LA-strain and 3D volume measurements of the left atrium can be useful parameters to use in detecting diastolic dysfunction in future recommendations. The result showed significant differences (p&lt;0.05) in LA-strain between normal diastolic function and grade 2 and -3. Also, between grade 1 and grade 2- and 3. Between undetermined diastolic function there was a statistical difference with grade 2- and 3. LA-strain showed a significant negative correlation between E/é and 2D. LA-strain and 3D-measurements of the left atrium can be useful tools in the next recommendations of grading diastolic dysfunction.

Neuroborrelios (NB) kan förekomma när Borrelia garinii, som är en del av artkomplexet Borrelia burgdorferi sensu lato (s.l), infekterar nervsystemet. När immuncellerna utsätts för borreliaantigen bildas antikroppar i plasma och intratekalt. NB ger även upphov till mononukleär pleocytos i cerebrospinalvätska (CSV). Neurologiska symptom, pleocytos och antikroppsindex (AI), som indikerar intratekal syntes av borreliaantikroppar, vägleder nuvarande NB-diagnostik på Klinisk Mikrobiologi i Kalmar. Diagnostiken försvåras vid tidig NB när AI är negativ och pleocytos positiv. För att komplettera tidig NB-diagnostik kan kemokinet C-X-C motif ligand 13 (CXCL13) mätas i CSV. I nuläget utförs CXCL13 enzymkopplad immunadsorberande analys (ELISA) på ett externt laboratorium. Mätning av CXCL13 kan även utföras med en snabb semi-kvantitativ ReaScan+ CXCL13 metod som bygger på lateral flödesimmunanalys (LFIA). Syftet var att verifiera ReaScan+ CXCL13, kontrollera linjäritet, CXCL13 stabilitet och bestämma diagnostisk specificitet och sensitivitet. CSV-proverna indelades i fyra grupper baserat på pleocytos och AI samt tre kategorier utifrån kriterier till NB-diagnos (n=31). Metodjämförelsen genomfördes mellan ReaScan+ LFIA och CXCL13 ELISA (n=25). Linjäritetsanalys utfördes på prov med hög CXCL13-koncentration i CSV och CXCL13-stabiliteten kontrollerades på två patientprover vid frys- och kylförvaring. Beräkning av sensitivitet och specificitet baserades på definitiv (n=20) och ej NB (n=5). Metodjämförelsen visade en fullständig överensstämmelse och Spearman´s korrelationskoefficienten var 0,949. Determinationskoefficienten för linjäritetsanalysen var 0,963. Diagnostisk sensitivitet var 80% och specificitet 100%. Hög prestanda, acceptabla stabilitets- och linjäritetsresultat samt ReaScan+ potentiella förmågan att särskilja mellan definitiv och ej NB, indikerar att metoden kan införas på Klinisk Mikrobiologi i Kalmar och komplettera nuvarande diagnostik av tidig NB. / Lyme neuroborreliosis (LNB) occurs when Borrelia garinii, part of the Borrelia burgdorferi sensu lato (s.l) species complex, infects the nervous system. Upon exposure, immune cells produce antibodies that can be measured in serum or cerebrospinal fluid (CSF). LNB also cause mononuclear pleocytosis in CSF. Neurological symptoms, pleocytosis and antibody index (AI), which indicate intrathecal synthesis of Borrelia antibodies, guide current diagnosis of LNB at the Clinical Microbiology in Kalmar. The diagnosis is complex in early LNB, especially when AI is negative and pleocytosis is positive. Expression of chemokine C-X-C motif ligand 13 (CXCL13) in CSF can be used as an additional marker to early LNB. Analysis is currently performed at an external laboratory using enzyme-linked immunosorbent assay (ELISA). The aim of this project was to evaluate the performance of the rapid semi-quantitative lateral flow immunoassay (LFIA) ReaScan+ CXCL13 test using 31 patient samples divided into four groups based on pleocytosis and AI. Linearity assay was performed on samples with high CXCL13 concentrations and stability was tested on two CSF samples after freezing and refrigeration. Estimation of sensitivity and specificity was based on definite (n=20) and non-LNB (n=5). Method comparison showed a complete agreement and Spearman's correlation coefficient was 0,949. The R2 for linearity was 0,963. Sensitivity was 80% and specificity 100%. Strong agreement, acceptable stability and linearity results as well as ReaScan+ potential ability to distinguish between definite and non-NB, indicate that the method can be introduced at Clinical Microbiology in Kalmar and used as a supplement in early LNB diagnostics.

In this study, the efficacy of MarrowMAX, a new IVD-certified culture medium for bone marrow samples, was evaluated and compared to the in-house modified culture medium RPMI-1640, which is routinely used at the Academic Hospital in Uppsala for chromosomal analysis. The hypothesis was that the new culture medium MarrowMAX would lead to an increased number of metaphases, resulting in a higher mitotic index, better spreading, and improved metaphase morphology. Twenty patient samples were cultured in parallel using both media and subjected to identical methods for sample preparation and staining for chromosomal analysis. The results demonstrated significant differences between MarrowMAX and RPMI-1640 for one-day, two-day, and three-day cultures. However, no significant differences were observed for directly harvested cultures and overnight colcemid cultures. These findings suggest that MarrowMAX is more effective than RPMI-1640, resulting in a higher frequency of metaphases, which is crucial for detailed chromosomal analysis. This was further confirmed by flow cytometry analysis, showing a higher number of proliferated cells in the MarrowMAX culture compared to the RPMI-1640 culture, in one representative sample. MarrowMAX's enhanced capacity to promote cell growth and structure in a laboratory setting indicates its potential as a promising alternative to RPMI-1640 for future diagnostic applications, potentially leading to faster diagnoses and improved treatment efficacy for patients with hematological conditions.

Efter upptäckten av penicillinet observerades en ökad resistens hos S. aureus vilket ledde till att alla stammar ansågs vara resistenta. Dock har många stammar enligt senare studier visat sig vara känsliga vilket har öppnat upp möjlighet att åter använda benzylpenicillin som ett behandlingsalternativ eftersom det kan ges i lägre koncentration jämfört med andra alternativ. Metoden för att upptäcka penicillinresistens hos S. aureus enligt EUCAST har bedömts svår att tillämpa på grund av subjektivitet vid bedömning av zonkanten. Studiens syfte var att utvärdera metoden för senare implementering på Klinisk Mikrobiologi Unilabs i Skövde. Metodens användbarhet utvärderades genom resistensbestämning av en referensstam samt tio stammar från ett referenslaboratorium för EUCAST. Bedömningen av de tio stammarna från referenslaboratoriet stämde vid jämförelse överens med deras resultat förutom för en stam som bedömdes som resistent i stället för känslig. Repeterbarheten var hög då ingen skillnad (p=0,156) uppvisades för referensstammen.  Resultatet tyder på att det kan vara svårt att göra en korrekt tolkning av zonkantens utseende då det är en subjektiv bedömning. För att uppnå en högre säkerhet kan det vara fördelaktigt att mer än en person gör en bedömning. Slutsatsen var att metoden kan implementeras på laboratoriet. / Following the discovery of penicillin, resistance rates increased until all strains of S. aureus were considered resistant. Recent studies have shown that many strains are susceptible, making it possible to again use penicillin as an alternative for treatment since it can be used in lower concentrations compared to alternatives. The EUCAST method for discovering penicillin resistance in S. aureus has been judged difficult due to the subjectiveness of assessing the zone edge. The aim of this study was to evaluate the method to implement it in the Section for Clinical Microbiology Unilabs Skövde. The useability of the method was evaluated by resistance testing of a reference strain and ten strains from a EUCAST reference laboratory. The assessment of the strains from the reference laboratory equaled their results except for one strain that was assessed resistant rather than sensitive. The repeatability was high since no difference (p=0,156) was found for the reference strain. The results show how difficult it is to correctly assess the zone edge due to its subjectiveness. It could be beneficial with more than one person assessing the zone edge to achieve a higher security. In conclusion the method is possible to implement in the laboratory.