At the histopathological laboratory at Ryhov County Hospital in Jönköping the dehydration process is currently scheduled during nights. To maintain good diagnostic quality with improved sample flow and use of the instruments full capacity an implementation of a shorter dehydration program scheduled during daytime was needed. The purpose was thus to evaluate a shorter dehydration program for punch biopsies of human skin and portio. The evaluation was based on ratings of cellular morphology and quality of routine staining with Hematoxylin and Eosin as well as special staining with polychrome methylene blue according to Unna, Periodic acid-Schiff and Giemsa. A material containing 87 biopsies was dehydrated distributed between current routine program and the evaluated shorter program. The specimens were rated on the basis of ability to section, morphological and staining quality along with diagnostic ability. The results of the survey showed that smaller punch biopsies run in the shorter dehydration program were easy to section and received good results in general. In contrast, larger biopsies were more difficult to section and showed more dehydration artefacts when dehydrated in the shorter program. Altogether it was concluded that the short dehydration program could be implemented for smaller punch biopsies from skin and portio at the histopathological laboratory at Ryhov County Hospital in Jönköping.

The assessment of right ventricular systolic function is done by echocardiography, with techniques as twodimensional echocardiography, motion-mode (M-mode) and Tissue Doppler Imaging (TDI). Measurements ​​obtained from these techniques are Tricuspid Annular Plane Systolic Excursion (TAPSE) and s`. TAPSE describes the longitudinal movement of annulus tricuspidalis. s´ describes annulus tricuspidalis maximal systolic velocity. The purpose of this essay was to explore the relationship between TAPSE and s´. The study consists of 99 participants between the ages of 18 and 90. The population was consisted of individuals with or without cardiovascular disease, but not individuals with atrial fibrillation. Participants were investigated with transthorakal echocardiography (TTE) at Länssjukhuset Ryhov in Jönköping. At the examination both TAPSE and s´ was measured. McNemar's test was performed and a Kappa value was calculated. The result showed a significant difference and the Kappa value was low which indicates a low relationship between the variables. The outcome of the study shows that several participants are classified with normal right ventricular function measured with TAPSE, but the same individuals are classified with right ventricular dysfunction measured by s'. The reason for the poor relationship between the parameters is unknown and should be studied further in future studies.

Anti-Müllerskt Hormon (AMH) är ett glykoprotein som spelar en avgörande roll för rätt behandling av patienter med fertilitetsproblem och diagnos av könsutvecklingsstörningar hos barn. Att utföra en verifiering av analys för AMH på sjukhuslaboratoriet i Värnamo, är av högsta relevans då analysen för närvarande inte utförs i region Jönköpings län. Repeterbarheten bedömdes genom 10 mätningar, medan total precision utvärderades med 4 dagliga mätningar under 5 dagar. Både repeterbarhet och total precision bedömdes med interna kontroller. Riktigheten inkluderade 22 patientprover från Linköping med varierande AMH-koncentrationer. Variationskoefficienten visade en mycket låg spridning kring medelvärdet för både repeterbarhet (1,22% och 0,75%) och total precision (1,39% och 1,12%). Resultatet på riktigheten visade ingen signifikant skillnad med ett p-värde på 0,061, vilket indikerar hög tillförlitlighet för klinisk användning och bidrar till att förbättra patientvården samt effektiviteten i behandlingen. Framtida studier kan dra nytta av att inkludera fler patientprover för att ytterligare utvärdera observerade skillnader.

Immunohistokemiska    (IHC) metoder används som diagnostiskt verktyg inom klinisk patologi. Många IHC-analyser ger information om sjukdomen och möjliga behandlingsalternativ. IHC-analys av HER2-proteinet är en vanligt prediktiv analys vid utredning av bröst- och ventrikeltumörer. Denna studie utfördes på avdelningen för Klinisk patologi på Södra Älvsborgs Sjukhus (SÄS). Syftet med studien var att utvärdera HercepTestTM mAb pharmDX (HercepTest (mAb)) för immunohistokemisk HER2-färgning av bröst- och ventrikelcancervävnad med avseende för ökad automatisering av laboratoriearbetet och förbättrat arbetsflöde. I studien jämfördes formalinfixerade paraffininbäddade preparat från bröst- (n=19) och ventrikelcancervävnad (n=12) färgade med det utvärderade reagenskittet mot färgning som hade utförts med HercepTestTM for Automated Link Platforms. Resultatet visade att HER2-färgning utfört med HercepTest (mAb) graderades likvärdigt eller högre på båda vävnadstyper och att diskordans mellan reagenskitten uppstod främst vid färgning av HER2-negativa prover. Möjliga orsaker till studiens resultat kan vara skillnaderna mellan färgningsprocessen och använt instrument för respektive reagenskit eller interpersonell variation i HER2-bedömning. Eventuella skillnader i reagenskittens förmåga att detektera HER2 är också en möjlig orsak till resultatet. Eftersom reagenskitten inte producerade likvärdiga resultat, avstod SÄS patologilaboratoriet att byta reagenskit för HER2-färgning och kommer troligtvis utföra vidare utvärderingar av analysen.

Bladder cancer is the seventh most common form of cancer in Sweden where the main source of material for investigation are bladder washings. A common method within liquid-based cytology for analysis of bladder washings is ThinPrep^®. Altered morphology as well as artifacts due to fixation can complicate funkar “the diagnostics of the sample. The aim of this study was to compare the slide content obtained from bladder washings fixed with fixative containing acetic acid with CytoLyt^® solution. The study also intended to compare the ThinPrep^® filters "Non-GYN" and "UroCyte". A total of 90 samples of bladder washings from patients where included. The samples where fixed with the addition of acetic acid and CytoLyt^®, using the "Non-GYN" and "UroCyte" filters. A cytotechnologist and a cytopathologist assessed the preparations based on the criteria background material, cell quantity and the amount of colour change. The results showed that CytoLyt^® in combination with the filter "Non-GYN" gave a more representative preparation and provided a good background image. The use of CytoLyt^® resulted in more diagnostic benefits compared to fixation with acetic acid, which also benefits patients.

The incidence of pancreatic ductal adenocarcinoma (PDAC) is increasing worldwide. The dismal prognosis and lack of effective treatments urges for increased research efforts in developing new treatment regimens. Since the development of new cancer treatments are expensive and time consuming, repurposing drugs is preferable when possible. This strategy will also be of great value for people in low income countries, to increase the availability of effective cancer treatments to affordable costs. Digitoxin, a cardiac glycoside, has been shown to have anti-cancer effects. It binds to the α subunit of the Na+/K+-ATPase, leading to increased concentrations of intracellular calcium and eventually cell death. There seems also to be other mechanisms elicited in cancer cells by digitoxin. The α subunit occurs in three isoforms, of which α3 has the highest affinity to digitoxin and frequently found over-expressed in tumor cells compared to normal cells.

PDAC cancer cells, both cell lines and tumors, differ in their genotype and in the metabolic subtype, proliferation rate and inflammatory status. To be able to individualize treatment regimens it is important to be aware of the specific vulnerabilities (genotypic or phenotypic characteristics increasing the sensitivity to digitoxin) of each PDAC tumor/cell line. This research aims to investigate the potential of using digitoxin as an anti-cancer treatment in PDAC, and analyze its effects on cell viability, metabolism and inflammatory status in PDAC cell lines in vitro with the goal to find biomarkers for digitoxin sensitivity.

The analyses of the effects of digitoxin was performed in five cell lines derived from PDAC tumors, either from primary tumors or metastases. Cell lines derived from PDAC are sensitive to digitoxin treatment to different degrees. High expression of α3 seems to be indicative for digitoxin sensitivity, as do a high proliferation rate seen in cell lines derived from primary tumors. Both subunit expression and proliferation rate should be further evaluated in PDAC tumors to confirm their potential to be used as biomarkers clinically.

In the hunt for the working mechanism behind the anti-cancer effects of digitoxin, the choline pathway, a pathway commonly affected in tumors was enlightened in the metabolomics study, affected in all five cell lines tested. Choline metabolites are important for maintaining the cell membrane and are involved in energy metabolism using lipids. Digitoxin induced an up-regulation of choline and glycerophosphocholine, which rendered us to propose a novel theory about possible interactions between two functional complexes in the cell membrane, the Na+/K+-ATPase/EGFR/c-Src and the EGFR/c-Src – Chkα. The hypothesis is that when digitoxin binds to the Na+/K+-ATPase it leads to inactivation of Chkα with a subsequent decrease in the synthesis of phosphocholine and phosphatidylcholine. Since cancer cells rely on abundance of choline metabolites, we believe an inhibition of this pathway to be deleterious for these cells.

Finally, we conclude that digitoxin has great potential as an anti-cancer treatment for some patients with pancreatic ductal adenocarcinoma. To optimize treatment results, a thorough investigation of the tumor genotype and phenotype must be done for each patient. To further increase treatment success, combination of digitoxin with other treatments for synergistic effects could be beneficial. 

Digitoxin has repeatedly shown to have negative effects on cancer cell viability; however, the actual mechanism is still unknown. In this study, we investigated the effects of digitoxin (1-100 nM) in four pancreatic cancer cell lines, BxPC-3, CFPAC-1, Panc-1, and AsPC-1. The cell lines differ in their KRAS/BRAF mutational status and primary tumor or metastasis origin. We could detect differences in the basal rates of cell proliferation, glycolysis, and ROS production, giving the cell lines different phenotypes. Digitoxin treatment induced apoptosis in all four cell lines, but to different degrees. Cells derived from primary tumors (Panc-1 and BxPC-3) were highly proliferating with a high proportion of cells in the S/G2 phase, and were more sensitive to digitoxin treatment than the cell lines derived from metastases (CFPAC-1 and AsPC-1), with a high proportion of cells in G0/G1. In addition, the effects of digitoxin on the rate of glycolysis, ROS production, and proliferation were dependent on the basal metabolism and origin of the cells. The KRAS downstream signaling pathways were not altered by digitoxin treatment, thus the effects exerted by digitoxin were probably disconnected from these signaling pathways. We conclude that digitoxin is a promising treatment in highly proliferating pancreatic tumors.

The alpha subunits (ATP1A1-3) of Na⁺/K⁺-ATPase binds digitoxin with varying affinity. The expression levels of these subunits dictate the anticancer effects of digitoxin. In the present study, three pancreatic cancer cell lines, AsPC-1, Panc-1 and CFPAC-1, were used to investigate the effects of digitoxin in relation to the expression of the subunits ATP1A1 and ATP1A3. Cell viability and intracellular calcium concentrations was measured in relation to the gene and protein expression of ATP1A1 and ATP1A3. Digitoxin was used to treat the cells at concentrations of 1-100 nM, and the intracellular calcium concentrations increased in a concentration-dependent manner in the Panc-1 and in the CFPAC-1 cells with treatment at 100 nM. In the AsPC-1 cells only the supraphysiological concentration of digitoxin (100 nM) resulted in a decrease in the number of viable cells (unviable cells increased to 22%), whereas it had no effect on intracellular calcium levels. The number of viable Panc-1 and CFPAC-1 cells decreased after digitoxin treatment at 25-100 nM (unviable Panc-1 cells increased to 33-59%; unviable CFPAC-1 cells increased to 22-56%). Digitoxin treatment also affected the transcriptional expression of the ATP1A1 and ATP1A3 subunits. In Panc-1 cells, ATP1A3 gene expression was negatively associated with the digitoxin concentration (25-100 nM). In the AsPC-1 and CFPAC-1 cells, the expression of the ATP1A1 gene increased in the cells treated with the 100 nM digitoxin concentration. The protein expression of ATP1A1 and ATP1A3 was not altered with digitoxin treatment. The basal protein expression of ATP1A1 was high in the AsPC-1 and CFPAC-1 cells, compared to the Panc-1 cells, in contrast to the basal expression of ATP1A3, which was higher in the Panc-1 cells, compared to the other pancreatic cancer cells used. On the whole, the present study demonstrates that the high expression of ATP1A3 renders pancreatic cancer cells more susceptible to digitoxin-induced cell death. The findings suggest that the expression of ATP1A3 may be used as a marker for tumor sensitivity to digitoxin treatment, where a high expression of ATP1A3 is favorable for the anticancer effects of digitoxin.

Elastic fibres ensure blood vessels and other tissues flexibility. Elastic staining of tissue is relevant when there is suspicion of melanoma, temporalis arteritis, venous invasion and after operations on blood vessels. The aim of the study was with the help of different tissue samples optimize and implement Hart´s elastic staining method as a substitute for Verhoeff’s at pathology lab at county hospital Ryhov, Jönköping. Colon, kidney, skin, and umbilical cord cross section got stained with Hart´s automated elastic staining method to evaluate the optimal staining procedure. Same region of the tissues was stained with Verhoeff´s manual elastic staining method and Hart´s method. All cross section were assessed and compared with the help of a pathologist doctor. Optimization of Hart´s method resulted in an incubation period of twelve minutes and optimal staining procedure without xylene. Result of comparison between Hart´s staining method and Verhoeff´s staining method showed that Hart´s staining method had a better contrast and background. Conclusions of the study was that Hart´s staining method was better than Verhoeff´s staining method, further studies could include research about a substitution of the blotting step with an extra ethanol bath as an example and liver tissue instead of the umbilical cord.

Diagnosis of vitamin B₁₂-defiency anemia has been challenging due to lack of ”gold standard” and poor sensitivity of total B₁₂ as a first line marker. With the aim of improving diagnostic of anemia investigations at county hospital Ryhov, Jönköping, we performed a method verification for active B12 on ARCHITECT i2000SR and evaluated the usefulness of active B₁₂ in diagnosing anemia. Correlation analysis was performed using samples with different total B₁₂ ranges: low (n=10, <148 pmol/L), grey zone (n=10, 149-200 pmol/L) and normal (n=10, 201-250 pmol/L). Receiver operating characteristic (ROC) curve was created and area under the curve (AUC) was calculated for determining accuracy of active B₁₂ using 14 deficiency cases. Coefficient of variation (CV) for within-run and total imprecision, for active B₁₂ was 2,4 % and 4,4 %, respectively. Weak correlation was found between B₁₂-ranges, however a moderate correlation (R=0,600) was found including all groups. Active B₁₂ with the cut-off value of 40 pmol/L resulted in high specificity (75%) and sensitivity (80%) and had an AUC of 90 %. Active B₁₂ assay on ARCHITECT i2000SR allows a precise measurement of active B₁₂ in human serum and therefore is adequate for routine use. Active B₁₂ had high accuracy and can be recommended in anemia investigations in county hospital Ryhov, Jönköping.

Stress stimulates SNS and the HPA-axis to release alpha-amylase and cortisol into the saliva. Alpha-amylase is analysed with enzyme kinetics and cortisol with ELISA.  There is no gold-standard for analysis with alpha-amylase and cortisol in saliva. The aim was to evaluate an analysis for alpha-amylase and cortisol with enzyme kits from Salimetrics and IBL International GMBH for future research studies. The study population consisted of 44 saliva samples that were collected 30 minutes after awakening before food intake. Two kits from respective company was used for each analyte in saliva. The activities of alpha-amylase and the concentrations of cortisol was measured spectrophotometric to compare the kits performance. The activity of alpha-amylase differed between Salimetrics (median=49,0 U/mL) and IBL International GMBHs (median=270,0 U/mL). The concentrations for cortisol differed slightly between Salimetrics (median=0,5 µg/dL) and IBL International GMBH (median=0,6 µg/dL). The correlation for alpha-amylase between the two kits were r=0,87,p>0,05 and  for cortisol, r=0,96,p>0,05. The difference in activity between the kits for alpha-amylase made it difficult to compare but Salimetrics results were easier to manage which made the kit more advantageously. The kits for cortisol showed similar concentrations but IBL International GMBH were more advantageously with regard for manage and price.

The Right ventricle (RV) has a complex anatomy, plays an important role in blood oxygenation and can be affected by several pathophysiological conditions. Evaluation of RV function has prognostic value in heart and lung diseases. Tricuspid annular plane systolic excursion (TAPSE) and annulus tricuspid peak systolic velocity (RVs´) are two common methods for assessing RV function in Transthoracic echocardiography. 

The study aimed to investigate interobserver variation, the correlation and agreement between TAPSE and RVs´. The study is a cross-sectional study of 53 healthy participants aged 18-60. The measurement was based on a blind double determination by two biomedical analyst students. The analysis was implemented with the program IBM SPSS Statistics.

Interobserver variation analysis showed no significant difference between the two students, (PTAPSE =0,568 and PRVs´=0,548). Regression analysis showed RVs´ had slightly less measurement uncertainty than TAPSE. A weak positive correlation was found between RVs´ and TAPSE and 100% agreement regarding outcome was demonstrated, Kappa value was 1. Both methods have good interobserver variation in inexperienced examiners. In people with healthy heart and lungs, the relationship between TAPSE and RVs looks weakly positive. Previous studies identified stronger positive association. Differences in the results may be due to a lack of experience on the part of the students and a low number of participants.