Background: Lactose intolerance (LI) is an inability or reduced ability to produce the enzyme lactase. This disorder causes discomfort like stomach pain and/or diarrhea. The most common cause of LI for adults is a mutation in the MCM6 gene, a single nucleotide polymorphism at C/T-13910. Haemochromatosis (HH) is a hereditary disease that causes more iron uptake from food than normal, which leads to more iron circulating in the blood and causing more iron to be stored in the liver. About 90% of HH patients are homozygous for C282Y and there is a connection between compound heterozygous for C282Y and H63D and potential development of HH. In this study, two different methods were used and compared to analyze the gene for LI (LCT) and the gene for HH (HFE). An extraction-free method by LaCAR and a DNA extraction In-house method done at Sundsvalls hospital Sweden. Method: Genotyping for LCT and HFE was done by polymerase chain reaction (PCR) technique on DNA extracted from whole blood (EDTA samples). Results: All the blood samples gave the same genotype on both methods. One LI sample had an extra curve at T/G-13915, which was detected by the LaCAR method but not the in-house method. One HH sample had a different C curve at 187, which was detected by both methods. Conclusion: The LaCAR method was identified as a good alternative for the analysis of LCT and HFE genotypes. It was able to detect additional mutations in the MCM6 gene for LCT analysis and has a shorter run time.

The antiseizure medication levetiracetam is used to treat epilepsy with significant success, the medication concentration in serum may be affected by other co-administered medication. Levetiracetam is excreted renally, and the halftime is depending on the renal function which is often correlated to age. The clinical chemistry laboratory at Sundsvall hospital did previously send samples for levetiracetam analysis to Karolinska University Hospital in Stockholm. There was a wish to start analysing the medication locally to reduce analysis time and thereby increase patient safety. ARK Diagnostic markets a kit for quantitative analysis of levetiracetam on automated chemistry analysers. The sample may be taken in a few different test tubes with slightly different characteristics. The aim of this study was to verify the ARK kit on Cobas Pro c503 at Sundsvall hospital and investigate the effects of different test tubes. To assess the accuracy and precision of the method, serum was spiked with levetiracetam from a liquid solution to construct dilution series for testing linearity and sample materials, and by running internal controls to assess the repeatability and reproducibility of the method. The results show that the method does in fact give accurate measurements of the levetiracetam concentration and the results does not vary more than acceptable between measurements nor over time and that different sample type does not show a clinically important difference in result.

Population numbers of bumblebees has been on a steady decline leading to the endangerment of several species that are critical to the growth and pollination of crops and plants. Varying factors have contributed to this situation, one key factor being pathogens wiping out colonies. Studying pathogens that diminish bumblebee populations is vital to understanding how to combat the decline of these important species effectively. The aim of this study was to analyse five of the most common pathogens that exist in the abdomen of five different species of bumblebees. This was achieved by extracting DNA, from 176 previously acquired frozen samples, through homogenisation and varying washing steps before an analysis of the resulting nucleic acids. The analysis was done with quantitative polymerase chain reactions to quantify the amounts of each pathogen, as well as a passive reference nucleic acid, to then be able to calculate the absolute total -amount of pathogen carried by each individual bumblebee. The results showed that almost half of the bumblebees were infected by Nosema, one sixth were infected by Crithidia and Apicystis while only a small number of individuals had been infected by Bombus Densovirus and the Apis Mellifera Filamentous Virus. Bombus Lapponicus was the most infected species. The conclusion based on the results is that future research could put a focus on the more commonly found parasites Nosema, Crithidia and Apicystis. Additional research should also be undertaken into the factors contributing to the less common pathogens Bombus Densovirus and the Apis Mellifera Filamentous Virus.

Amyloidosis is a group of diseases that originate from misfolded proteins. These proteins form fibrils which is stored in the tissue and organs. Approximately three to twelve per million people are diagnosed with amyloid light chain immunoglobulin amyloidosis (AL) each year and the median age is 65. The prognosis for AL amyloidosis is poor with a median survival rate of six months to three years. The patients may survive an additional 15-20 years with treatment such as stem cell transplantation.   The aim of this rapport was to optimize a monoclonal antibody named pwlam to diagnose AL λ amyloid with immunohistochemistry (IHC) when the amount of sample is small. In this rapport three different methods were used, pressure preparation, IHC and western blot. For both pressure preparation and IHC, colour development occurred in case of positive results. Western blot was used as a quality control for the tissue since they were all originally diagnosed using that method.    A total of 26 samples were tested, 16 positive AL λ samples, five AL κ samples and five negative samples. Pwlam is developed against AL λ so the AL κ samples function as a quality control for the antibody since it doesn’t bind to the κ light chains. A total of seven AL λ samples had a good binding with pwlam. More research is needed before pwlam can be used in IHC on pressure preparations as a supplementary method to diagnose samples when the amount of AL λ amyloid is small.

Laktatdehydrogenas (LD) är en vanlig biomarkör som används för att uppskatta allmän vävnadsskada och celldöd i kroppen. Det mäts i blodserum och analyseras, liksom de flesta andra blodprover, så snart som möjligt efter korrekt förberedelse. Det kan dock uppstå förseningar i analysen som leder till att analyser måste senareläggas vilket i sin tur kan leda till nedbrytning av vissa molekyler och kemiska strukturer i prover. Studien syftade till att undersöka om, och i så fall i vilken omfattning, nedbrytning på grund av förvaring i rumstemperatur (22 °C) respektive kyla (4 °C) som påverkar de kvantifierbara resultaten av LD inom 30 timmar efter att proverna tagits. För att uppnå detta togs blodprover (n = 30) från inneliggande patienter där serum alikvoterades till två rör där ena röret förvarades i rumstemperatur och andra i kylskåp varpå upprepade mätningar av LD utfördes under totalt 26 timmar. Resultaten visade att förvaringstemperatur och tid inte hade någon statistiskt signifikant påverkan på värdet av LD (p > 0,05) och att LD därmed kan mätas oavsett förvaring i kyla eller rumstemperatur inom 29 timmar efter provtagningen.

Background Multiple myeloma is an incurable cancer disease that emerges from the bone marrow. Bone morphogenetic proteins (BMPs) are ligands that activates intracellular signaling pathways causing activation of transcription factors. Previous studies show that BMP treatment of myeloma cells induce apoptosis, a mechanism dependent on downregulation of c-MYC. BMPs uses different receptors on myeloma cells, but it is still unclear how the intracellular signaling pathway leading to apoptosis works. A recent whole genome CRISPR/Cas9 knockout screening suggested FOXO1 as a gene involved in the mechanism of apoptosis during BMP treatment. We therefore aimed to investigate further on how FOXO1 has an impact on BMP induced apoptosis. Methods Our hypothesis was that knockout of FOXO1 would protect the cells from apoptosis. To begin to address this issue we tested INA-6 FOXO1 knock-out cell clones that was generated before the start of the project and treated them with BMP-9 to look for effects on cell viability and protein expression. We measured cell viability using CellTiter-Glo® 2.0 Cell viability assay and expression of c-MYC and FOXO1 protein using Western blot. Results and conclusions Treatment with BMP-9 for 72 hours showed a decrease in viability of the cells, up to 98%. Protein expression of c-MYC was inhibited by BMP-9 treatment while a constant expression of FOXO1 was seen in all cells clones regardless of BMP treatment. Expression of FOXO1 in the FOXO1 knock-out cells indicates that the knock-out has not worked. More experiments are needed to clarify the role of FOXO1 in BMP-induced apoptosis.

Background: Resistance against antibiotics is a global problem that can lead to serious complications. Many bacteria have developed resistance against several antibiotics which causes infections that are hard to treat. Resistant bacteria such as Methicillin-resistant Staphylococcus aureus (MRSA) and Vancomycin-resistant Enterococcus (VRE) are mainly spread within healthcare. To combat these infections, it is important to have reliable detection methods. Purpose: To qualitatively compare the Copan Transsystem 114c with theESwab-system for the detection of MRSA and VRE. Methods: For the study, Polymerase chain reaction (PCR) methods, Matrix-assisted laser desorption/ionization Time of Flight (Maldi-Tof), and microbiological cultivation methods was used on simulated and clinical samples. Results: For the VRE-samples no difference in results were identified. The MRSA samples showed significant differences in PCR-results where the Transsystem 114c detected 16/60 positive samples whereas the ESwab-system detected 12/60. But the cultivation results showed opposite results where the Transsystem 114c detected three positive samples and the ESwab-system detected four. Conclusion: The results show that both the Transsystem 114c and ESwab show similar results on simulated samples but differ on clinical samples. Further comparative tests need to be carried out to validate the methods before they can be used in routine operation thus no real conclusion can be drawn from this study.

No description available.

No description available.

Abstract Addressing the formidable challenges presented by diffuse midline glioma (DMG), a notably aggressive pediatric brain tumor with limited therapeutic options, this study investigates the oncogenic roles of MYC and PDGFB and evaluates SOX9's contribution to therapeutic resistance. Utilizing advanced transgenic mouse models and the RCAS virus system, our research effectively simulates the progression and treatment responses characteristic of DMG.   In our approach, we established a DMG tumor model by manipulating Tumor Virus A receptor (TVA)-expressing embryonic neural stem cells sourced from the E12.5 hindbrain in vitro. Validation of this model and its genetic perturbations was achieved through detailed Western blot analyses. Results indicate that DMG cells overexpressing MYC tended to be significantly more sensitive to chemotherapy compared to PDGFB cells that do not overexpress MYC, positioning MYC as a crucial therapeutic target. On the other hand, increased expression of SOX9 was linked to a slight increase in resistance to conventional treatment modalities, highlighting its role in promoting adaptive resistance mechanisms within the tumor microenvironment.   This research emphasizes the critical importance of merging molecular biology techniques with comprehensive in vivo modeling to elucidate DMG's pathophysiology and identify actionable therapeutic targets. Our findings offer significant insights into the molecular dynamics of DMG and suggest novel targets for therapeutic intervention that could substantially improve clinical outcomes in this challenging pediatric malignancy. Future research should aim to broaden the genetic profiling of DMG, and tailor therapeutic approaches based on specific molecular profiles to optimize treatment efficacy and precision.