Analysis of stroma-derived factors with potential prognostic and therapeutic significance in prostate cancer

Prostate cancer is the most common malignancy in Sweden with about 9000 new cases diagnosed every year. New markers are needed to improve diagnostic accuracy. The most commonly used tissue-biomarkers are basal cell markers and AMACR, often used in combination. We identified three potential tissue biomarkers, CYCS, ICK and IKBKB, by using the Human Protein Atlas database and investigated their diagnostic accuracy. The potential of these biomarkers was also compared with AMACR. Immunohistochemical analysis of the markers was performed on a tissue microarray (TMA) consisting of tissue from 40 prostate specimens, including benign prostatic tissue, atrophy, high-grade prostatic intraepithelial neoplasia (HGPIN) and prostate cancer. In addition, qRT-PCR analysis of malignant and benign frozen tissue samples from 32 radical prostatectomy specimens was performed. All four biomarkers showed a higher protein expression in prostate cancer and HGPIN than in benign tissue. The prognostic accuracy was highest for AMACR, but the results indicate that in some cases CYCS, ICK and IKBKB may serve as additional diagnostic markers. It is known that prognostic information can also be derived from tumor stroma. The prognostic value of stromal expression of PDGFRβ was therefore evaluated by immunohistochemical analysis of PDGFRβ on a TMA containing cancer and non-malignant tissue from more than 300 prostate cancer patients. The association between stromal staining intensity and a number of clinical characteristics were then analyzed. Expression of PDGFRβ in non-malignant and tumor stroma was associated with high Gleason grade and reduced cancer specific survival. Cancer associated fibroblasts (CAFs) are found in many solid tumors and promote tumor growth and progression. Identification and inhibition of molecules mediating these interactions constitute an attractive strategy for development of new cancer therapies. By comparative analyses of CAFs and normal fibroblasts from prostate tissue we have identified a number of genes upregulated in prostate CAFs. CXCL14, an orphan chemokine, was the most upregulated transcript. Overexpression of CXCL14 in fibroblasts increased their proliferation and migratory capacity. Also over-expression in fibroblasts of CAF led to increased ability of these cells to stimulate proliferation and migration of prostate cancer cells. Furthermore, fibroblasts overexpressing CXCL14 enhanced tumor growth, vascularisation and macrophage infiltration in a stroma-dependent prostate cancer model. Another transcript identified to be upregulated in prostate CAFs was GDF15, a member of the TGFβ superfamily. GDF15 was shown to stimulate fibroblast proliferation and enhanced growth, migration and invasion of prostate cancer cells. Fibroblasts over-expresssing GDF15 was also able to stimulate prostate xenograft growth when co-injected with prostate cancer cells. Interestingly, these fibroblasts also increased the ability of tumor xenograft to promote growth at a distant site suggesting direct or in-direct systemic pro-tumoral effects of fibroblast-derived GDF15. These studies thus identify a set of new diagnostic and prognostic markers for prostate cancer and stromaderived potential therapeutic targets

How is the hind leg temperature, circumference and diameter affected by different types of stable bandages?

Tidigare studier visar att ledproblem var ett vanligt förekommande problem hos sporthästar. En av dessa ledproblem som hästen kan lida av är svullnader i lederna på hästens ben. En svullnad i hästens leder kan uppkomma på grund av ökad ledvätska i leden och det är den typen av svullnad som med talspråk ofta kallas för galla. Svullnaden kan vara ofarlig för hästen men det kan också vara en indikation på ledproblem som bör uppmärksammas av hästägaren. Svullnaden i benen kan också uppkomma efter intensiv och upprepad träning. Syftet med studien är att jämföra tre behandlingsmetoder för svullnad i hästens ben; två metoder med olika typer av bandageunderlägg och en behandling där hästen står utan bandageunderlägg. Det besvarades genom frågeställningarna; Kan olika typer av bandageunderlägg minska gallor i hästens kotled och kotsenskida? Kan olika typer av bandageunderlägg påverka hudens temperatur på hästens bakben vid kotled och kotsenskida? Studien är en kvantitativ studie och utformades som en change-over design med washout period av ett dygn mellan mätningar. Studien pågick under elva dagar och innefattade nio varmblodshästar (7–24 år) från Ridskolan Strömsholm. Resultatet visade att Back on Track® och ordinärt bandageundelägg hade en signifikant skillnad (p=0,002) i temperatur (mätt med termistor), diameter kotled (mätt med skjutmått) och omkrets kotled (mätt med måttband). Medelvärdet för temperaturen för Back on track® och ordinärt bandageunderlägg var 34.3±0.31C, för diameter kotled 85.1±0.25 mm (Back on Track®) och 85.0±0.25 mm (ordinärt bandageunderlägg) och för omkrets 31.2±0.11 cm (Back on track®) och 31.1±0.11 cm (ordinärt bandageunderlägg). Slutsatsen som kunde dras var att med ett bandageunderlägg minskar omfång i diameter och omkrets vid kotled och kotsenskida och ger en förhöjd temperatur i huden på benen. Vilket material som används till ett bandageunderlägg har ingen signifikant påverkan på varken diameter, omkrets eller temperatur på benet.Previous studies show that joint problems were a common problem in sport horses. One of these joint problems that the horse may suffer from is swelling of the joints on legs. A swelling of the joints can occur due to increased joint fluid in the joint. The swelling can be harmless to the horse, but it can also be an indication of joint problems that should be brought to the attention of the horse owner. The swelling of the legs can also occur after intense and repeated exercise. The aim of the study is to compare three treatment methods for swelling of the horse's legs; two methods with different types of wool bandages and a treatment where the horse is without wool bandages. It was answered through the questions; Can different types of stable bandages reduce bile in the horse's vertebrae and vertebral sheath? Can different types of stable bandages affect the skin's temperature on the horse's hind legs at the fetlock joint? The study was a quantitative study and was designed as a Change-over design with a washout period of one day between measurements. The study lasted for eleven days and included nine warmblood horses (7–24 years) at the riding School Strömsholm. The result showed that Back on Track® and ordinary bandage had a significant difference (P=0.002) in temperature (measured with thermistor), Diameter fetlock joint (measured by caliper) and circumference fetlock joint (measured by tape measure). The mean temperature for Back on track® and ordinary bandage was 34.3±0.31C, for diameter fetlock joint 85.1±0.25 mm (Back on Track®) and 85.0±0.25 mm (ordinary bandage) and for circumference 31.2±0.11 cm (Back on track) and 31.1±0.11 cm (ordinary bandage). The conclusion was that wool bandages reduce the circumference and in the diameter at the ancle joint and vertebral sheath and gives an increased blood circulation which is shown by elevated temperature in the leg. The material used for stable bandages had no significant effect on either the diameter, circumference, or the temperature of the leg

iNKT cell regulation of B cell activation in inflammation

The ability to combat infections through the generation of specific immune responses is critical to our survival. The immune system can react to and combat virtually any molecule presented to it, including those derived from our own body. The immune system can cause considerable damage to the structures it was designed to protect, thus causing autoimmune disorders. Examples include systemic lupus erythematosus, Sjögren’s syndrome, and rheumatoid arthritis, among many others. Many autoimmune disorders are caused in part by autoreactive antibodies produced by B lymphocytes, and targeting the activation of B lymphocytes thus forms a target for ameliorating disease. Better understanding of how B lymphocytes are activated is essential not only in order to limit autoimmune disease, but also in order to harness their capabilities in the context of generating successful vaccines. The work presented in this thesis deals with B lymphocyte activation in inflammation and in response to glycolipid antigens. The aim was to investigate how this activation is regulated and investigate the possibility of manipulating B cell activation in a desirable way. In papers I and III, mouse models were used to study the regulation of autoreactive and IgE-producing B lymphocytes using IL-18 to induce inflammation. In paper II, model glycolipid antigens were employed to investigate how different types of iNKT cell help affect the outcome for B cell activation, with the prospect of harnessing iNKT cell help as an adjuvants in vaccine therapy. In paper IV, the potential of using glycolipids in order to skew faulty B cell activation in inflammation was investigated. In paper I, iNKT cells were identified as regulators of autoreactive and IgE-producing B cells. Paper II defines quantitative and qualitative differences between cognate and noncogante B cell help provided by iNKT cells. Cognate iNKT cell help, compared to noncognate, expands a larger number of B cells producing the cytokine Interleukin 10. In paper III, mechanisms of iNKT cell regulation of B cell activation were investigated. Neutrophils were found to license iNKT cells to adopt a killing phenotype required to restrict B cell activation in sterile inflammation. In paper IV, B cell activation in inflammation was studied using exogenous glycolipid iNKT cell antigens. Serum antibody levels and germinal center formation were found to be increased in inflammation when iNKT cells were stimulated with glycolipids. In summary, the work presented in this thesis describes mechanisms controlling faulty B cell activation in inflammation. iNKT cells are critical in limiting B cell activation, and this was dependent on interaction with neutrophils. The interaction between neutrophils and iNKT cells plays a previously unappreciated role in the restriction of B cell activation, and thus serves as a potential target for new therapeutic strategies in autoimmune diseases. In addition, the use of glycolipids as vaccine adjuvants, or to target B cell activation is investigated

Hur påverkas bakbenets temperatur, omkrets och diameter av olika typer av bandageunderlägg?

Tidigare studier visar att ledproblem var ett vanligt förekommande problem hos sporthästar. En av dessa ledproblem som hästen kan lida av är svullnader i lederna på hästens ben. En svullnad i hästens leder kan uppkomma på grund av ökad ledvätska i leden och det är den typen av svullnad som med talspråk ofta kallas för galla. Svullnaden kan vara ofarlig för hästen men det kan också vara en indikation på ledproblem som bör uppmärksammas av hästägaren. Svullnaden i benen kan också uppkomma efter intensiv och upprepad träning. Syftet med studien är att jämföra tre behandlingsmetoder för svullnad i hästens ben; två metoder med olika typer av bandageunderlägg och en behandling där hästen står utan bandageunderlägg. Det besvarades genom frågeställningarna; Kan olika typer av bandageunderlägg minska gallor i hästens kotled och kotsenskida? Kan olika typer av bandageunderlägg påverka hudens temperatur på hästens bakben vid kotled och kotsenskida? Studien är en kvantitativ studie och utformades som en change-over design med washout period av ett dygn mellan mätningar. Studien pågick under elva dagar och innefattade nio varmblodshästar (7–24 år) från Ridskolan Strömsholm. Resultatet visade att Back on Track® och ordinärt bandageundelägg hade en signifikant skillnad (p=0,002) i temperatur (mätt med termistor), diameter kotled (mätt med skjutmått) och omkrets kotled (mätt med måttband). Medelvärdet för temperaturen för Back on track® och ordinärt bandageunderlägg var 34.3±0.31C, för diameter kotled 85.1±0.25 mm (Back on Track®) och 85.0±0.25 mm (ordinärt bandageunderlägg) och för omkrets 31.2±0.11 cm (Back on track®) och 31.1±0.11 cm (ordinärt bandageunderlägg). Slutsatsen som kunde dras var att med ett bandageunderlägg minskar omfång i diameter och omkrets vid kotled och kotsenskida och ger en förhöjd temperatur i huden på benen. Vilket material som används till ett bandageunderlägg har ingen signifikant påverkan på varken diameter, omkrets eller temperatur på benet.Previous studies show that joint problems were a common problem in sport horses. One of these joint problems that the horse may suffer from is swelling of the joints on legs. A swelling of the joints can occur due to increased joint fluid in the joint and it is the type of swelling, this swelling is also called bile. The swelling can be harmless to the horse, but it can also be an indication of joint problems that should be brought to the attention of the horse owner. The swelling of the legs can also occur after intense and repeated exercise. The aim of the study is to compare three treatment methods for swelling of the horse's legs; two methods with different types of wool bandages and a treatment where the horse is without wool bandages. It was answered through the questions; Can different types of stable bandages reduce bile in the horse's vertebrae and vertebral sheath? Can different types of stable bandages affect the skin's temperature on the horse's hind legs at the fetlock joint? The study was a quantitative study and was designed as a Change-over design with a washout period of one day between measurements. The study lasted for eleven days and included nine warmblood horses (7–24 years) at the riding School Strömsholm. The result showed that Back on Track® and ordinary bandage had a significant difference (P=0.002) in temperature (measured with thermistor), Diameter fetlock joint (measured by caliper) and circumference fetlock joint (measured by tape measure). The mean temperature for Back on track® and ordinary bandage was 34.3±0.31C, for diameter fetlock joint 85.1±0.25 mm (Back on Track®) and 85.0±0.25 mm (ordinary bandage) and for circumference 31.2±0.11 cm (Back on track) and 31.1±0.11 cm (ordinary bandage). The conclusion was that wool bandages reduce the circumference and in the diameter at the ancle joint and vertebral sheath and gives an increased blood circulation which is shown by elevated temperature in the leg. The material used for stable bandages had no significant effect on either the diameter, circumference, or the temperature of the leg

Efficacy and safety of atomoxetine as add-on to psychoeducation in the treatment of attention deficit/ hyperactivity disorder

Objective: The primary objective of this study was to assess the impact of atomoxetine in combination with psychoeducation, compared with placebo and psychoeducation, on health-related quality of life (HRQL) in Swedish stimulant-naïve pediatric patients with attention deficit/ hyperactivity disorder (ADHD). HRQL results will be presented elsewhere. Here, psychoeducation as well as efficacy and safety of the treatment are described. Patients and methods: A total of 99 pediatric ADHD patients were randomized to a 10-week double-blind treatment with atomoxetine (49 patients) or placebo (50 patients). Parents of all patients received four sessions of psychoeducation. Atomoxetine was dosed up to approximately 1.2 mg/kg day (≤70 kg) or 80 mg/day (>70 kg). Improvement of ADHD symptoms was evaluated using the ADHD rating scale (ADHD-RS) and clinical global impression (CGI) rating scales. Safety was assessed based on adverse events (AEs). Results: The study population was predominantly male (80.8%) and diagnosed with the combined ADHD subtype (77.8%). The least square mean (lsmean) change from baseline to endpoint in total ADHD-RS score was −19.0 for atomoxetine patients and −6.3 for placebo patients, resulting in an effect size (ES) of 1.3 at endpoint. Treatment response (reduction in ADHD-RS score of ≥25 or ≥40%) was achieved in 71.4 or 63.3% of atomoxetine patients and 28.6 or 14.3% of placebo patients. The lsmean change from baseline to endpoint in CGI-Severity was −1.8 in the atomoxetine group compared with −0.3 in the placebo group. The difference between treatments in CGI-Improvement at endpoint was −1.4 in favor of atomoxetine. No serious AEs occurred. The safety profile of atomoxetine was in line with the current label. Conclusions: Atomoxetine combined with psychoeducation was superior to placebo and psychoeducation in ADHD core symptoms improvement. The large ES might be a result of including stimulant-naïve patients only, but also may indicate a positive interaction between atomoxetine treatment and psychoeducation, possibly by increased compliance

Stromal PDGFRβ Expression in Prostate Tumors and Non-Malignant Prostate Tissue Predicts Prostate Cancer Survival

Background: The identification of new prognostic markers for prostate cancer is highly warranted, since it is difficult to identify patients requiring curative treatment. Data from both experimental models and clinical samples have identified important functions of PDGFRb on pericytes and fibroblasts in the tumor stroma. Methodology/Principal Findings: In this study the prognostic significance of PDGFRb in prostate cancer stroma, and in matched non-malignant tissue, was evaluated with immunohistochemistry. PDGFRb expression was analyzed in normal and tumor stroma from more than 300 prostate cancer patients. High PDGFRb expression in tumor stroma was associated with large tumor size, advanced stage, high Gleason score and high vessel density. Perivascular PDGFRb staining in tumors was also correlated with high Gleason score. Correlations were also observed between PDGFRb status in tumor stroma and nonmalignant stroma. Similarly, high PDGFRb expression in adjacent non-malignant tissue stroma correlated with large tumor size, advanced stage, high Gleason score and proliferation in non-malignant epithelium. Interestingly, high levels of PDGFRb in the stroma of tumor and non-malignant tissue were associated with shorter cancer specific survival in prostate cancer patients. Conclusions/Significance: The study revealed a number of novel associations between stromal PDGFRb expression i

Structures of Human Antibodies Bound to SARS-CoV-2 Spike Reveal Common Epitopes and Recurrent Features of Antibodies

Neutralizing antibody responses to coronaviruses mainly target the receptor-binding domain (RBD) of the trimeric spike. Here, we characterized polyclonal IgGs and Fabs from COVID-19 convalescent individuals for recognition of coronavirus spikes. Plasma IgGs differed in their focus on RBD epitopes, recognition of alpha- and beta-coronaviruses, and contributions of avidity to increased binding/neutralization of IgGs over Fabs. Using electron microscopy, we examined specificities of polyclonal plasma Fabs, revealing recognition of both S1^A and RBD epitopes on SARS-CoV-2 spike. Moreover, a 3.4Å cryo-EM structure of a neutralizing monoclonal Fab-spike complex revealed an epitope that blocks ACE2 receptor binding. Modeling based on these structures suggested different potentials for inter-spike crosslinking by IgGs on viruses and that characterized IgGs would not be affected by identified SARS-CoV-2 spike mutations. Overall, our studies structurally define a recurrent anti-SARS-CoV-2 antibody class derived from VH3-53/VH3-66 and similarity to a SARS-CoV VH3-30 antibody, providing criteria for evaluating vaccine-elicited antibodies

Atomoxetine improves patient and family coping in attention deficit/hyperactivity disorder: a randomized, double-blind, placebo-controlled study in Swedish children and adolescents

This 10-week study assessed the efficacy of atomoxetine in combination with psychoeducation compared to placebo and psychoeducation in the improvement of Quality of Life in Swedish stimulant-naive children and adolescents with attention deficit/hyperactivity disorder. A total of 99 patients were treated with atomoxetine (49 patients) or placebo (50 patients) for 10 weeks and assessed regarding broader areas of functioning using the Quality of Life measures Child Health and Illness Profile-Child Edition (CHIP-CE), Family Strain Index [FSI; equivalent to the Family Burden of Illness Module used in the study], Appraisal of Stress in Child-Rearing (ASCR), Five to fifteen (FTF), “I think I am” (“Jag tycker jag är”), and Children’s Depression Rating Scale-Revised (CDRS-R) before and after the active treatment phase. Simultaneously, the patients’ parents participated in a 4-session psychoeducation program. A statistically significant difference in favor of atomoxetine was seen in the improvement from baseline to study endpoint for the CHIP-CE domains “Achievement” and “Risk avoidance”, for the FSI total score, for the ASCR section (I) domain “Child as a burden”, for all FTF domains except for “Language and Speech”, and for the CDRS-R total score. No difference between treatment groups was observed in the patient-assessed evaluation of self-esteem using the “I think I am” scale. Atomoxetine combined with psychoeducation had a positive effect on various everyday coping abilities of the patients as well as their families during 10 weeks of treatment, whereas the patients’ self-image and the parents’ image of the climate in the family were not significantly improved