No description available.

No description available.

Background: Idiopathic inflammatory myopathies (IIM) constitute a heterogeneous group of diseases with severe consequences for the life of affected patients. Dermatomyositis, polymyositis and inclusion body myositis (IBM) are the classical representatives of this group. The treatments given today often have limited effects, and are taken at the cost of side effects. Major obstacles in the search for more effective treatments are; (1) an incomplete understanding of the disease mechanisms, (2) difficulties to delineate homogeneous disease groups for clinical studies and (3) the sometimes challenging task to diagnose these diseases. Aims: We addressed a number of “loose ends” in the areas of pathogenesis, classification and diagnosis; mechanisms of muscle fiber degeneration in IIM, with a focus of programmed cell death (apoptosis) and invasion of muscle  fibers by inflammatory cells (partial invasion); protecting and mediating factors present in muscle; the association of other diseases with IIM, in particular celiac disease ; the evaluation of two classification systems and laboratory methods for increased diagnostic performance. The studies: We included 106 patients, diagnosed at the Neuromuscular unit in Linköping, Sweden, with pathological muscle findings consistent with IIM. The incidence in the county of Östergötland (during 5 years) was 7.3 per million/year (3 patients each year). Of 88 patients with confirmed IIM 4 (4.5 %) had celiac disease, 33 (38%) had an associated systemic inflammatory disease and 5 (5.7 %) had a malignancy. Ninety-nine patients were included for a comparison of two classification systems using criteria of the European Neuromuscle Centre (Amato/ENMC), and the widely used Bohan and Peter classification, both with the addition of IBM according to Griggs et al. Using the Amato/ENMC criteria the most prevalent diagnostic group after IBM (30%) was nonspecific myositis (23%), followed by polymyositis (20%) and dermatomyositis 17%). A substantial number of patients meeting Bohan and Peter (or Griggs) criteria were excluded by Amato/ENMC criteria, most (21/23) due to lack of detectable muscle weakness. Extended muscle sectioning increased the sensitivity of a muscle biopsy by 15 % and the specificity by 22%, and showed an overlap between disease groups. Muscle biopsies from patients with IIM and controls were used to investigate pathological findings considered specific for disease groups, and for the presence of programmed cell death (apoptosis) and disease protecting and mediating factors in muscle. The presence of apoptotic muscle fiber nuclei was detected in muscle with partial invasion (however not in the invaded fibers) in the presence of granzyme B and CD8+ cytotoxic T cells. The major apoptosis inhibiting protein Bcl-2 was shown to be constitutionally expressed in healthy muscle but weakened in IIM. Conclusion: We present apoptosis as a possible disease mechanism in parallel with partial invasion of fibers. Furthermore, partial invasion may not be a suitable distinguishing feature in the pathogenesis, or for classification and diagnosis of IIM. We also introduce the anti-apoptotic Bcl-2 as a possible relevant muscle fiber protecting factor. A more extensive pathological work-up improves classification and diagnosis of IIM. The proposed Amato/ENMC creates a substantial portion of patients with non-specific or unclassified myositis. Associated diseases are common in IIM, and also include celiac disease.

No description available.

Enligt Socialstyrelsens föreskrifter om ledningssystem för kvalitet och patientsäkerhet i hälso- och sjukvården, SOSFS 2005:12, krävs det av vårdgivaren att denne arbetar efter mål. Att formulera mål inom hälso- och sjukvården kräver eftertanke. Målen måste vara mätbara och inkludera flera olika delar av verksamheten. Varje förbättringsprojekt är unikt och bör relateras till de resurser som avsatts. Ett stort problem vid kontinuerlig uppföljning är att välja ut parametrar som ger tillräcklig information och är lätta att skapa. Här krävs mer utvecklade datasystem och även integrering mellan befintliga datasystem  I uppsatsen diskuteras med vilka parametrar man kan utvärdera ett förbättringsprojekt inom laboratoriemedicin. Det finns för närvarande inte några standardiserade kvalitetsparametrar inom klinisk kemi. De parametrar som i denna undersökning visade sig vara mest lämpliga att använda kontinuerligt i det korta perspektivet var svarstider, personalresurser och reagenskostnader. I det längre perspektivet kan ovanstående data kombineras till indikatorer som visar kostnad per analyspoäng och analyspoäng per årsarbetare. De indikatorer som kan få användning först när datasystem utvecklas är data från avvikelsesystem och analyskommentarer som visar t.ex. att svar inte har kunnat lämnas ut. / According to the National Board of Health and Welfare, SOSFS 2005:12, it is required that the caregiver adheres to his work through following stated objectives. Formulating goals in health care requires careful consideration. Goals must be measurable and include many different aspects of the project. Each improvement project is unique and should be directly related to the resources allocated to the project. A major problem for continuous monitoring is to select parameters that provide sufficient information and are easy to produce. This requires more advanced computer systems and integration with existing computer systems. The paper discusses the parameters an improvement project in laboratory medicine can evaluate its results after. There are currently no standardized quality parameters in clinical chemistry. The parameters used in this study that proved to be the most suitable to be used continuously in the short term was the response times, staffing and reagent costs. In the long term, the above data are combined with indicators that show cost per analysis score and analysis points per full-time employees. The indicator that may be used when the computer system is developed is deviation systems and analytical comments that show for example that a response has not been disclosed.

Primary aldosteronism (PA) is a common form of secondary hypertension with an international prevalence rates between 5 and 10 %. It is characterized by a high autonomous aldosterone production that causes cardiovascular damage, renin suppression, hypertension, sodium retention, potassium excretion and hypokalemia. The screening of PA is a simple test measuring aldosterone to renin ratio (ARR) with immunoassay method. This test is currently considered as the most reliable screening tool for PA.     The main objective of the study was to evaluate an ELISA-method, for detection of aldosterone and renin in blood plasma, to be used for routine analysis in the laboratory. The second aim was to investigate the effect of refreezing samples, considering that cryoactivation of prorenin might occur.     One hundred blood samples were analysed, in regard to aldosterone and renin, by using two commercial ELISA assays (DRG ELISA from DRG Diagnostics, Germany) on a Dynex DS2 instrument. In addition, the accuracy and precision of the methods were calculated. The effect of refreezing was investigated with a series of eight samples, which were analyzed twice on the same instrument.     Both assays performed well. The resulting data showed good precision and accuracy. The correlation between the original and refreezed samples was good, r = 0.989 and r = 1.0 for aldosterone and renin respectively. Considering that the study only included eight samples, further investigation is recommended.     Evaluation showed that both immunoassays are reliable in diagnostic use and the ELISA-method is suitable to implement in the laboratory for routine analysis.

Avsaknad av en ”gyllene standard” och en låg sensitivitet hos förstahandsmarkören total B12 har varit de stora begränsningarna i diagnostiken av vitamin B12-brist anemier. Med syftet att förbättra diagnostiken av bristanemier vid Länssjukhuset Ryhov, Jönköping utvärderades användningen av aktivt B12 vid bristanemiutredningar och en metodverifiering för analysen genomfördes på ARCHITECT i2000SR. Sambandet mellan aktivt B12 och totalt B12 undersöktes med en korrelationsanalys för totalt B12-nivåer: låg (n=10, <148 pmol/L), gråzonen (n=10, 149–200 pmol/L) och normal (n=10, 201-250 pmol/L). För bestämning av träffsäkerheten hos aktivt B12 studerades 14 bristanemifall. Ett receiver operating characteristic (ROC) diagram skapades och arean under kurvan (AUC) beräknades. Variationskoefficient (CV) för mellanliggande- och inomserieprecision för aktivt B12 var 4,4 % respektive 2,4 %. Svag korrelation observerades i alla B12-nivåer, men sammantaget resulterade nivåerna i en medelstark korrelation (R=0,600). Aktivt B12 med ett gränsvärde på 40 pmol/L resulterade i en hög specificitet (75 %) och sensitivitet (80 %) med en AUC på 90 %. Analys av aktivt B12 på ARCHITECT i2000SR möjliggör en precis mättning av aktivt B12 i humant serum och därför är lämplig för rutinmässig användning. Träffsäkerheten hos aktivt B12 var hög och markören kan därmed rekommenderas för användning vid bristanemiutredningar vid Länssjukhuset Ryhov, Jönköping. / 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.

Duktal bröstcancer är den vanligaste formen av invasiva brösttumörer. Graderingssystemet för bröstcancer har definierats av Elston och Ellis och är baserat på tre parametrar. En av dessa är räkning av antal mitoser på preparat färgade med Hematoxylin och Eosin (HE). Som ett komplement vid bedömning av bröstcancer analyseras Ki67, en proliferationsmarkör, med hjälp av immunhistokemi (IHC).  Fosfohiston H3 (pHH3) är ett histonprotein som finns i cellkärnan. Proteinet tros vara en specifik markör för mitos eftersom den är fosforylerad enbart under M-fasen och i slutet av G2-fasen. Syftet med studien var att utvärdera pHH3 som potentiell markör för mitos vid diagnostiseringen av duktal bröstcancer. Syftet var även att jämföra metoden med räkning av antal mitoser och Ki67-positiva celler, samt att studera den interindividuella skillnaden vid bedömningen av preparaten. Materialet bestod av 20 sektorresektat med invasiv duktal bröstcancer. Preparaten färgades med IHC och bedömdes mikroskopiskt. Celler som var positiva för pHH3 och Ki67 samt antal mitoser räknades, av tre läkare. Ett medelvärde för varje patientfall och metod beräknades från läkarnas bedömningar. Metodernas variationskoefficienter och dess medelvärden beräknades. Variationskoefficienterna uppvisade medelvärden på 0,21 för Ki67 +/- 0,10 SD, 0,33 för pHH3 +/- 0,14 SD och 0,46 för mitosräkning +/- 0,34 SD. Korrelationskoefficienterna för metoderna och respektive läkare uppvisade en spridning. Korrelationerna uppvisade medelvärden på r = 0,78 för Ki67 och pHH3, r = 0,74 för Ki67 och mitos samt r = 0,83 för pHH3 och mitos. Enligt studien verkar antipHH3 vara ett bra komplement vid bedömning av duktal bröstcancer. Dock krävs tydliga kriterier för vad som ska räknas som en pHH3-positiv cell. Intervariabiliteten verkar bli mindre med anti-pHH3 än vid räkning av mitoser, som är mer tidskrävande. Minst intervariabilitet ses vid bedömning av anti-Ki67 som en proliferationsmarkör. / Ductal carcinoma of the breast is the most common form of invasive breast tumours. The grading system for breast cancer is defined by Elston and Ellis and is based on three criterions. One of these criterions is the mitotic count in pathological sections of breast carcinomas stained with Hematoxylin Eosin. A common method often applied as a complement in diagnosis of breast carcinoma is immunohistochemical staining with use of antibodies directed against Ki67, a proliferation marker. Phosphohistone H3 is a histone protein that is located in the cell nucleus. The protein is believed to be a specific marker for mitosis since it only is phosphorylated during mitosis, and to some extent at the end of the G2-phase. The purpose of this study was to evaluate pHH3 as a potential marker for mitosis when diagnosing ductal breast cancer. The purpose was also to compare the method to mitotic figuring and the count of Ki67-positive cells, and to study the inter-individual variability when assessing the histological sections. The material consisted of 20 biopsies containing invasive ductal breast cancer. The sections were stained using IHC and all sections were evaluated microscopically. Cells positive for pHH3, Ki67 and mitotic cells were quantified, by three doctors. From the doctors results an average value was determined for each case and method. To be able to compare the methods the coefficient of variation was calculated. The average value of the coefficient of variation was determined for each method and also the standard deviation (SD). The coefficient of variation showed average values of 0,21 for Ki67 +/- 0,10 SD, 0,33 for pHH3 +/- 0,14 SD and 0,46 for mitotic figuring +/- 0,34 SD. The correlation coefficients for the methods and each doctor showed dispersion. The correlations showed average values of r = 0,78 for Ki67 and pHH3, r = 0,74 for Ki67 and mitosis and r = 0,83 for pHH3 and mitosis. According to this study it seems as though anti-pHH3 could complement the other methods. However explicit criteria which defines a threshold value of which cells should be considered pHH3-positive needs to be established. The inter-individual differences seem to decrease using antipHH3 compared with mitotic counting, which is more time consuming. Although the minimum difference can be seen when assessing anti-Ki67 as a proliferation marker.

Procalcitonin (PCT) is a precursor protein of the hormone calcitonin and is encoded by the gene Calcitonin-1. In the Blekinge Regional, P-PCT is only analyzed in Karlskrona. The analysis is performed in the department of clinical chemistry on the Alinity i-series instrument. PCT indicates bacterial infections and therefore, it is important to have a backup method for the analysis when the instrument in the city of Karlskrona is out of order. The aim of this work was to verify the analysis method of P-PCT on the instrument Alinity i-series in the city of Karlshamn. Analysis method verification means to confirm and prove that the method meets the specified requirements. Verification was performed by analyzing 35 samples with different concentration of PCT on the master instrument in Karlskrona and on “Alinity 1” and “Alinity 2” in Karlshamn. The method was compared by studying correlation coefficient and bias. The precision was measured only on “Alinity 1” which would be the master instrument in Karlshamn. Precision was measured by analyzing 25 replicates at two control levels and then was 5 replicates of each control level analyzed over 5 days. The correlation was good and no significant bias between results from Karlskrona and “Alinity 1” and between results from “Alinity 1” and “Alinity 2”. Precision on “Alinity 1” meets the requirements. The conclusion was that verification of PCT on master instrument “Alinity 1” and slave instrument “Alinity 2” was approved and the backup method for the PCT analysis in Karlshamn was verified. / Prokalcitonin (PCT) är en peptidprekursor till hormonet kalcitonin och kodas av genen kalcitonin-1 (CALC-1). PCT används som biomarkör för bakteriella infektioner och sepsis. I Region Blekinge analyseras P-PCT bara i Karlskrona. Analysen utförs på avdelningen för Klinisk kemi på instrumentet Alinity i-serie Abbot. PCT indikerar bakteriella infektioner och av denna anledning efterfrågas av flera avdelningar. Därför är det viktigt att verifiera en analysmetod för P-PCT i Karlshamn, ifall instrumenten i Karlskrona är ur funktion. Syftet med arbetet var att verifiera analysmetoden för P-PCT på instrumentet Alinity i-serie Abbot i Karlshamn. Metodverifiering innebär att bekräfta och bevisa att metoden uppfyller specificerade krav. Metodverifiering utfördes genom att analysera 35 prover med olika PCT-koncentrationer på masterinstrument ”Shrek” i Karlskrona och på ”Alinity 1” och ”Alinity 2” i Karlshamn. Metoderna jämfördes genom att studera korrelationskoefficient och bias. Inom och total serie-precision mättes bara på ”Alinity 1” som ska vara masterinstrument i Karlshamn. Inom serie-precision mättes genom att analysera 25 replikat av två kontrollnivåer och total serie-precision genom att analysera 5 replikat/dag under 5 dagar. Resultaten blev att metoden på ”Alinity 1” hade en god korrelation med metoden på masterinstrument ”Shrek” och ingen signifikant bias observerades mellan metoderna. Inom och total serie-precision på ”Alinity 1” uppfyller kraven. Metoden på ”Alinity 2” hade en god korrelation med metoden på masterinstrument ”Alinity 1” och ingen signifikant bias observerades mellan metoderna. Slutsatsen var att verifiering av P-PCT på masterinstrument ”Alinity 1” och slavinstrument ”Alinity 2” var godkänd och därmed verifierades en backupmetod för analysen P-PCT i Karlshamn.

Neuroblastoma constitutes roughly 8% of all childhood cancers where 95% of all neuroblastoma cases occur before the age of 10. The survival rate of infants and young children is very poor, which alone contributes to research novel biomarkers for classification methods, improved diagnosis and better anti-tumor therapies. The aim of this meta-analysis was to identify dysregulated miRNAs in neuroblastoma that has the potential to be used as antioncogenic biomarkers for diagnostic interventions. Additionally, explore miRNA interconnectedness on a systemic level and conversely extend the support of using miRNAs as biomarkers. A comprehensive literature search was performed within NIH-PubMed, NCBI-PMC and in the reference list of already reviewed publications, which yielded 9 eligible publications. Quality of evidence was assessed according to the guidelines adapted from MIAME, MINSEQE and MIQE. miRNet 2.0 was used to find the most significantly enriched annotations linked to neuroblastoma. A total of 251 samples (Cancer: 141; Control: 110) was reported by the 9 studies. These involved 66 dysregulated miRNAs (Up-regulated: 43; Down-regulated: 23) which was used for enrichment analysis. Four miRNAs (miR-17-5p, -92a-3p -421, -125b) were significantly linked to neuroblastoma, and associated secondary diseases; medulloblastoma (-92a-3p, -125b), bladder cancer (-17-5p, -125b), acute myeloid leukemia (-92a-3p, -125b) and cardiac hypertrophy (- 125b). miR-125b showed exceptional interconnectivity with these diseases and a multidimensional potential in neural tumorigenesis. This study showed that dysregulation and biological processes of these miRNAs were concurrent with the original studies, endorsing that these miRNAs have potential as diagnostic indicators or classifiers of such diseases. / Popular scientific summary Neuroblastoma (NB) is one of the most common types of pediatric neurological cancers in children and constitutes roughly 8% of all childhood cancer types, in which 95% of all NB cases occur before the age of 10. Even with frequent advancements in medical diagnosis and anti-tumor therapies, the current treatment options for patients with NB offers a survival rate that is very poor. This alone is a reason to pursue developing novel classification methods, improve diagnosis and research better anti-tumor therapies. Micro Ribonucleic Acids (miRNAs) are small non-coding single stranded biomolecules that have gotten a lot of attention in recent years due to their ability to regulate genes involved in various biological cancer processes, such as; tumor growth and development. miRNAs regulate these processes by altering the function of messenger RNAs (mRNAs), which are single-stranded biomolecules that resembles a piece of genetic code from the DNA of an organism cell. When these mRNAs become dysregulated, their cancer-promoting genes are disrupted which prevent them from working properly, leading to tumor regression or termination. The effect of this biological event is then objectively measured by using the miRNA as an indicator, also known as a biomarker. miRNA biomarkers have massive potential to improve various medical applications, such as; faster and more accurate diagnosis, detailed disease-classification and more precise drug trial predictions. However, a lot of individual studies have been published about the same miRNAs, which report a variation of conclusions. This makes it more difficult to determine the true nature of miRNAs. This issue can be addressed with systematic reviews and meta-analyses, which could yield additional support and give a broader picture of how miRNAs regulate different biological processes in NB. A meta-analysis is a scientific statistical process that combines the results of many research publications associated with the same scientific question and presents the best collective estimate of truth with increased precision than what could be achieved from individual studies alone. Thus, meta-analysis is an important tool in research which makes sure that the most trustworthy effect estimate can be achieved among many similar answers. The aim of this meta-analysis was to identify dysregulated miRNAs in NB that has the potential to be used as anti-cancer promoting biomarkers for diagnostic interventions. Additionally, explore how different miRNAs are connected to NB and conversely extend the support of using miRNAs as biomarkers. The end goal of this meta-analysis is to provide more reliable evidence for further research that can improve the life expectancy of NB patients in the future. In this study, 4 miRNAs (miR-17-5p, -92a-3p -421 and -125b) were identified to be significantly linked to NB, and associated secondary diseases; medulloblastoma (-92a-3p & -125b), bladder cancer (-17-5p & -125b), acute myeloid leukemia (-92a-3p & -125b) and cardiac hypertrophy (- 125b). Specifically, miR-125b showed exceptional interconnectivity for these diseases and potential to indirectly down-regulate n-Myc in NB, a gene that promote cancer cell proliferation. miR-125b was also found to be a significant sole regulator and effector of the CDX2 gene responsible for cancer cell differentiation in acute myeloid leukemia, a relationship that has been supported by other publications. This meta-analysis showed that the reported dysregulation and biological processes of these miRNAs were concurrent with the original studies, endorsing that these miRNAs have potential as diagnostic indicators or classifiers of such diseases while warranting that the gene regulatory function of miRNAs are becoming more intricate than previously thought.