Human epididymis protein 4 reference limits and natural variation in a Nordic reference population

The objectives of this study are to establish reference limits for human epididymis protein 4, HE4, and investigate factors influencing HE4 levels in healthy subjects. HE4 was measured in 1,591 samples from the Nordic Reference Interval Project Bio-bank and Database biobank, using the manual HE4 EIA (Fujirebio) for 802 samples and the Architect HE4 (Abbott) for 792 samples. Reference limits were calculated using the statistical software R. The influence of donor characteristics such as age, sex, body mass index, smoking habits, and creatinine on HE4 levels was investigated using a multivariate model. The study showed that age is the main determinant of HE4 in healthy subjects, corresponding to 2% higher HE4 levels at 30 years (compared to 20 years), 9% at 40 years, 20% at 50 years, 37% at 60 years, 63% at 70 years, and 101% at 80 years. HE4 levels are 29% higher in smokers than in nonsmokers. In conclusion, HE4 levels in healthy subjects are associated with age and smoking status. Age-dependent reference limits are suggested

An Experimental Study of Vibration Assisted Displacement in a Horizontal Annulus for Enabling Through Tubing Abandonment

Traditionally, Permanent Plug and Abandonment (PP&A) operations has been carried out by pulling casing and tubing using either Mobil Offshore Drilling Unit (MODU) or Fixed Platform Drilling (FPD). The costs can run up to 25% of the total well, therefore, new cost-efficient PP&A technology is a necessity (Khalifeh & Saasen, 2020b). In later years Through- Tubing Abandonment (TTA) has been viewed as a good alternative, by not having to pull any casing or tubing and thus saving rig time. This method can also be used on a Light Well Intervention Vessel (LWIV) to further reduce the costs significantly (Thom, Angell, Greig, Robertson, & Hogg, 2020). However, a challenge with this method that causes great concern, is whether the drilling fluid will be properly displaced and removed in the annulus due to lack of tubing centralization and possibly unfavorable flow dynamics. To study how the TTA method can be improved, this thesis aim to investigate the effect of vibration at different frequencies and eccentricities in laboratory experiments using a vibration tool, with a particular focus on enabling TTA. The experiments was carried out using a test rig built at Norwegian Research Centre (NORCE). The test rig is a downscaled annulus made out of a steel tubing and a transparent casing and fitted with a vibration motor for moving the inner tubing. In these experiments, flow rate, densities and viscosities of the displacing and displacement fluids were designed by the use of dimensional analysis to reflect a real-life operation. These parameters were kept constant while eccentricity and vibration was varied to investigate their effect on the displacement process. The allocated laboratory time period was unfortunatly shortened due to time constraint caused by the COVID-19 pandemic. A total of 26 test runs was carried out, where 12 test runs was used in this study. The findings in these experiments indicates that gravity is the dominating force for this system, consequently resulting in an undesired slumping displacement front. Nevertheless, as eccentricity increases, the gravitational force becomes less dominant and a more even and steady piston like displacement front is observed. Moreover, the results indicates that high vibration will promote fluid to flow in the entire area of the annulus; both on the upper wide part and the lower narrow part. This results in a more evenly distributed and steadier displacement front. But in this study, the low- and medium- vibrations does not appear to significantly improve the displacement efficiency. In spite of this, it is noteworthy to mention that vibration has been found to be a useful tool for decreasing undesirable high yield- and gel strengths in old drilling fluid (Thom et al., 2020)

Effects of hypoxia on angiogenesis and angiogenic factors in crucian carp brain

Most vertebrates depend on an uninterrupted supply of oxygen to maintain the energy production. Some species, like the crucian carp (Carassius carassius) can survive hypoxia for an extended period. This fish is an expert in acquiring the little oxygen that exists in hypoxic water, and can boost its anaerobic ATP production by up-regulating the glycolysis and convert lactate to ethanol. Angiogenesis, the formation of new blood vessels, is induced by hypoxia and controlled through several factors. One of the most important hypoxia driven transcription factors, the hypoxia-inducible factor (HIF)-a, has been found to play a major role in coordinating many adaptive responses to hypoxia. It is continuously synthesized and degraded in normoxic conditions, but accumulates at low oxygen tension. Several factors control the expression of HIF-a, among them the factor-inhibiting HIF-1 (FIH-1). One mechanism by which HIF-a mediates increased oxygen delivery is through inducing the expression of vascular endothelial growth factor (VEGF), which in turn stimulates the formation of blood vessels. In this thesis mRNA levels of HIF-1a, HIF-2a, FIH, VEGFA, VEGFC and VEGFD were quantified by real-time RT-PCR in brain tissue from the crucian carp exposed to hypoxia. The effect of hypoxia on vascular density in crucian carp brain was also studied. A significant increase in mRNA levels was seen in both VEGFA and VEGFC, while no change was seen in VEGFD. VEGFA increased 6-fold and constituted the largest part of the quantified VEGF expression. Different splice isoforms of VEGFA have previously been characterized in mammals and fish, and two of these splice isoforms were cloned in the crucian carp, VEGFA121 and VEGFA165. A trend towards a reduction in VEGFD expression was observed in hypoxia, which may indicate that it does not have the same function in angiogenesis in crucian carp as VEGFA and VEGFC. The expression level of FIH increased 4-fold in hypoxia. No changes in mRNA levels were seen in either HIF-1a or HIF-2a, but a trend towards reduction was seen in HIF-1a. The increase seen in VEGFA and FIH indicates that the fishes have been hypoxic. This might imply that the expression of HIF-a also increases, but in order to verify this protein analyses would have to be performed to verify this. Regarding blood vessel density two parameteres were measured, blood vessel surface area per unit volume and blood vessel length per unit volume. A significant decrease was seen in both of these parameters. The increase in VEGFA expression along with a reduction in blood vessel density can indicate that the hypoxic crucian carp does not require additional blood vessels during hypoxia, but instead suppresses global HIF-a effects, which may work well for local regulation of vascularization, but not for adapting the whole organism to hypoxia

An Experimental Study of Vibration Assisted Displacement in a Horizontal Annulus for Enabling Through Tubing Abandonment

Traditionally, Permanent Plug and Abandonment (PP&A) operations has been carried out by pulling casing and tubing using either Mobil Offshore Drilling Unit (MODU) or Fixed Platform Drilling (FPD). The costs can run up to 25% of the total well, therefore, new cost-efficient PP&A technology is a necessity (Khalifeh & Saasen, 2020b). In later years Through- Tubing Abandonment (TTA) has been viewed as a good alternative, by not having to pull any casing or tubing and thus saving rig time. This method can also be used on a Light Well Intervention Vessel (LWIV) to further reduce the costs significantly (Thom, Angell, Greig, Robertson, & Hogg, 2020). However, a challenge with this method that causes great concern, is whether the drilling fluid will be properly displaced and removed in the annulus due to lack of tubing centralization and possibly unfavorable flow dynamics. To study how the TTA method can be improved, this thesis aim to investigate the effect of vibration at different frequencies and eccentricities in laboratory experiments using a vibration tool, with a particular focus on enabling TTA. The experiments was carried out using a test rig built at Norwegian Research Centre (NORCE). The test rig is a downscaled annulus made out of a steel tubing and a transparent casing and fitted with a vibration motor for moving the inner tubing. In these experiments, flow rate, densities and viscosities of the displacing and displacement fluids were designed by the use of dimensional analysis to reflect a real-life operation. These parameters were kept constant while eccentricity and vibration was varied to investigate their effect on the displacement process. The allocated laboratory time period was unfortunatly shortened due to time constraint caused by the COVID-19 pandemic. A total of 26 test runs was carried out, where 12 test runs was used in this study. The findings in these experiments indicates that gravity is the dominating force for this system, consequently resulting in an undesired slumping displacement front. Nevertheless, as eccentricity increases, the gravitational force becomes less dominant and a more even and steady piston like displacement front is observed. Moreover, the results indicates that high vibration will promote fluid to flow in the entire area of the annulus; both on the upper wide part and the lower narrow part. This results in a more evenly distributed and steadier displacement front. But in this study, the low- and medium- vibrations does not appear to significantly improve the displacement efficiency. In spite of this, it is noteworthy to mention that vibration has been found to be a useful tool for decreasing undesirable high yield- and gel strengths in old drilling fluid (Thom et al., 2020)

Successful triple immuno-enzymatic method employing primary antibodies from same species and same immunoglobulin subclass

Protocols for immunohistochemical (IHC) detection of multiple antigens in the same tissue sections have been developed using primary antibodies directly conjugated to different enzymes or fluorochromes, or ones that have been raised in different species, or from different immunoglobulin (Ig) classes or subclasses. For antibodies lacking such dissimilarities, very few proposals have been published with varying degrees of generalizability. In this report we present a successful triple IHC protocol engaging three unconjugated monoclonal primary antibodies raised in the same species and of the same Ig subclass. Compared to other methods, our results showed that denaturation of the preceding reaction complex by microwave heating, combined with additional suppression of enzyme activity, enabled the detection of all three reactions by using the same detection system, with no cross reaction observed. Moreover, expression patterns of each of the three antigens in the triple stained sections, was found to be similar to the pattern observed when single staining was performed. Unlike previous reports, no damage of targeted antigens or tissues did occur following this protocol. Furthermore, the contrast of the colors employed was investigated by computerized color deconvolution, and the three reactions products were successfully separated into three individual images that could be used for further objective quantification

Utbygging av digitalt bakkenett i Norge : NRK og TV 2s motiver

Målet med utredningen er å analysere NRK og TV 2s motiver ved utbyggingen av det digitale bakkenettet. Vi identifiserer både mediepolitiske- og økonomiske motiver. I forhold til medievitenskap finner vi at hensynet til allmennkringkasting vil være sentralt for både NRK og TV 2, blant annet fordi man bruker allmennkringkastingskravene til å posisjonere seg i markedet. De økonomiske motivene har sin bakgrunn i teori om bedriftservervelse og vi identifiserer både ledelsesmotiver og profittmotiver. Ulike profittmuligheter undersøkes videre ved hjelp av teori for vertikale relasjoner, og vi finner at integrasjonen mellom NRK og TV 2 vil kunne øke profitten blant annet ved eliminering av eksternaliteter. Det er dog problematisk å prise innholdstilbudet i fjernsynsmarkedet og potensialet for profitt vil i stor grad knytte seg til tilleggstjenester. Det er vanskelig å konkludere med ett hovedmotiv, men det er ikke tvil om at NRK og TV 2 vil styrke sin posisjon i det norske fjernsynsmarkedet og kanskje vil overføre sin produsentmakt videre til distribusjonsleddet

Salivary glands of primary Sjögren’s syndrome patients express factors vital for plasma cell survival

Introduction: The presence of circulating Ro/SSA and La/SSB autoantibodies has become an important marker in the classification criteria for primary Sjögren’s syndrome (pSS). Plasma cells producing these autoantibodies are mainly high affinity plasma cells originating from germinal centre reactions. When exposed to the right microenvironment these autoimmune plasma cells become long-lived and resistant to immunosuppressive treatment. Since autoimmune plasma cells have been detected in the salivary glands of SS patients, we wanted to investigate if the glandular microenvironment is suitable for plasma cell survival and if glandular residing plasma cells are the long-lived plasma cell subset. Methods: Single, double and triple immunohistochemistry as well as immunofluorescence staining was performed on minor salivary gland tissue retrieved from pSS, chronically inflamed and normal subjects. Results: We detected significant numbers of CD138+, non-proliferating, Bcl-2 expressing plasma cells in the salivary glands of pSS patients with high focus score (FS). Furthermore, we demonstrated that CXCL12 and interleukin (IL)-6 survival factors were highly expressed in pSS salivary gland epithelium and by focal mononuclear infiltrating cells. Notably, adipocytes when present in the salivary gland tissue were an important source of CXCL12. We clearly demonstrate that plasma cells are localised in close proximity to CXCL12 and IL-6 expressing cells and thus that the environment of salivary glands with high FS provide factors vital for plasma cell survival. Conclusions: Plasma cells residing in the salivary glands of pSS patients with high FS showed phenotypic characteristics of the long-lived plasma cell subtype. Furthermore, the pSS salivary gland microenvironment provided niches rich in factors vital for plasma cell survival

Specific antibodies and sensitive immunoassays for the human epidermal growth factor receptors (HER2, HER3, and HER4)

The use of trastuzumab in patients with breast cancer that overexpresses human epidermal growth factor receptor 2 has significantly improved treatment outcomes. However, a substantial proportion of this patient group still experiences progression of the disease after receiving the drug. Evaluation of the changes in expression of the human epidermal growth factor receptors could be of interest. Monoclonal antibodies against the extracellular domain of the human growth factor receptors, 2, 3, and 4, have been raised, and specific and sensitive immunoassays have been established. Sera from healthy individuals (Nordic Reference Interval Project and Database) were analyzed in the human epidermal growth factor receptor 2 assay (N = 805) and the human epidermal growth factor receptor 3 and 4 assays (N = 114), and reference limits were calculated. In addition, sera from 208 individual patients with breast cancer were tested in all three assays. Finally, the human epidermal growth factor receptor 2 assay was compared with a chemiluminescent immunoassay for serum human epidermal growth factor receptor 2/neu. Reference values were as follows: human epidermal growth factor receptor 2, <2.5 µg/L; human epidermal growth factor receptor 3, <2.8 µg/L; and human epidermal growth factor receptor 4, <1.8 µg/L. There were significant differences in human epidermal growth factor receptor 2 and human epidermal growth factor receptor 3 serum levels between the patients with tissue human epidermal growth factor receptor 2–positive and tissue human epidermal growth factor receptor 2–negative (p = 0.0026, p = 0.000011) tumors, but not in the serum levels of human epidermal growth factor receptor 4 (p = 0.054). There was good agreement between the in-house human epidermal growth factor receptor 2 assay and the chemiluminescent immunoassay. Our new specific antibodies for all the three human epidermal growth factor receptors may prove valuable in the development of novel anti-human epidermal growth factor receptor targeted therapies with sensitive immunoassays for measuring serum levels of the respective targets and in monitoring established treatment

Vibration-assisted annular fluid displacement for rig-less well abandonment operations

Through-tubing abandonment is the operation where the production tubing is cut or punched and cemented inside the production casing. This can establish a full cross-sectional barrier or isolation in wells where the cement behind the production casing provides zonal isolation. Furthermore, isolation in the form of a through-tubing abandonment plug can be placed from a light-well intervention vessel, which reduces the risk and carbon footprint compared to conventional abandonment designs that rely on a full mobile offshore drilling unit. A potential challenge involved in placing the through-tubing abandonment plug is the risk of incomplete fluid displacement from the annulus behind the tubing: The tubing eccentricity may be considerable where the barrier should be placed, and the original packer fluid may be contaminated resulting in unfavorable displacement conditions. Furthermore, the tubing cannot be reciprocated or rotated to compensate for these conditions. Agitator tools that generate lateral and axial vibrations in the tubing for mitigating potentially adverse displacement conditions have recently become available to the industry. We study potential effects of such tools by performing annular displacement experiments using a novel test rig that enables lateral movement of the inner tubing. The original fluid to be displaced was viscosified with a polymer additive to generate apparent yield stress behavior and to thereby develop challenging displacement conditions, particularly in the eccentric annulus. Experimental results and supporting numerical simulations show that lateral motion of the tubing is effective in displacing the yield stress fluid from the narrow side of the eccentric annulus, and to oppose the tendency toward stratification in both concentric and eccentric annuli. The experiments further show that vibration is effective in improving the displacements at inclinations ranging from vertical to 80° from the vertical, and that high frequency vibrations are more effective than lower frequency vibrations at the same amplitude. Results from a mechanical vibration analysis suggests the vibration amplitudes increase with increasing flow rates, and that potentially beneficial vibrations can propagate over axial lengths that are comparable or longer than minimum barrier length requirements