Development and validation of a porcine artificial colonic mucus model reflecting the properties of native colonic mucus in pigs

Colonic mucus plays a key role in colonic drug absorption. Mucus permeation assays could therefore provide useful insights and support rational formulation development in the early stages of drug development. However, the collection of native colonic mucus from animal sources is labor-intensive, does not yield amounts that allow for routine experimentation, and raises ethical concerns. In the present study, we developed an in vitro porcine artificial colonic mucus model based on the characterization of native colonic mucus. The structural properties of the artificial colonic mucus were validated against the native secretion for their ability to capture key diffusion patterns of macromolecules in native mucus. Moreover, the artificial colonic mucus could be stored under common laboratory conditions, without compromising its barrier properties. In conclusion, the porcine artificial colonic mucus model can be considered a biorelevant way to study the diffusion behavior of drug candidates in colonic mucus. It is a cost-efficient screening tool easily incorporated into the early stages of drug development and it contributes to the implementation of the 3Rs (refinement, reduction, and replacement of animals) in the drug development process

Boceprevir for untreated chronic HCV genotype 1 infection.

International audienceBACKGROUND: Peginterferon-ribavirin therapy is the current standard of care for chronic infection with hepatitis C virus (HCV). The rate of sustained virologic response has been below 50% in cases of HCV genotype 1 infection. Boceprevir, a potent oral HCV-protease inhibitor, has been evaluated as an additional treatment in phase 1 and phase 2 studies. METHODS: We conducted a double-blind study in which previously untreated adults with HCV genotype 1 infection were randomly assigned to one of three groups. In all three groups, peginterferon alfa-2b and ribavirin were administered for 4 weeks (the lead-in period). Subsequently, group 1 (the control group) received placebo plus peginterferon-ribavirin for 44 weeks; group 2 received boceprevir plus peginterferon-ribavirin for 24 weeks, and those with a detectable HCV RNA level between weeks 8 and 24 received placebo plus peginterferon-ribavirin for an additional 20 weeks; and group 3 received boceprevir plus peginterferon-ribavirin for 44 weeks. Nonblack patients and black patients were enrolled and analyzed separately. RESULTS: A total of 938 nonblack and 159 black patients were treated. In the nonblack cohort, a sustained virologic response was achieved in 125 of the 311 patients (40%) in group 1, in 211 of the 316 patients (67%) in group 2 (P<0.001), and in 213 of the 311 patients (68%) in group 3 (P<0.001). In the black cohort, a sustained virologic response was achieved in 12 of the 52 patients (23%) in group 1, in 22 of the 52 patients (42%) in group 2 (P=0.04), and in 29 of the 55 patients (53%) in group 3 (P=0.004). In group 2, a total of 44% of patients received peginterferon-ribavirin for 28 weeks. Anemia led to dose reductions in 13% of controls and 21% of boceprevir recipients, with discontinuations in 1% and 2%, respectively. CONCLUSIONS: The addition of boceprevir to standard therapy with peginterferon-ribavirin, as compared with standard therapy alone, significantly increased the rates of sustained virologic response in previously untreated adults with chronic HCV genotype 1 infection. The rates were similar with 24 weeks and 44 weeks of boceprevir

Frequency drift in MR spectroscopy at 3T

Purpose: Heating of gradient coils and passive shim components is a common cause of instability in the B-0 field, especially when gradient intensive sequences are used. The aim of the study was to set a benchmark for typical drift encountered during MR spectroscopy (MRS) to assess the need for real-time field-frequency locking on MRI scanners by comparing field drift data from a large number of sites.Method: A standardized protocol was developed for 80 participating sites using 99 3T MR scanners from 3 major vendors. Phantom water signals were acquired before and after an EPI sequence. The protocol consisted of: minimal preparatory imaging; a short pre-fMRI PRESS; a ten-minute fMRI acquisition; and a long post-fMRI PRESS acquisition. Both pre- and post-fMRI PRESS were non-water suppressed. Real-time frequency stabilization/adjustment was switched off when appropriate. Sixty scanners repeated the protocol for a second dataset. In addition, a three-hour post-fMRI MRS acquisition was performed at one site to observe change of gradient temperature and drift rate. Spectral analysis was performed using MATLAB. Frequency drift in pre-fMRI PRESS data were compared with the first 5:20 minutes and the full 30:00 minutes of data after fMRI. Median (interquartile range) drifts were measured and showed in violin plot. Paired t-tests were performed to compare frequency drift pre- and post-fMRI. A simulated in vivo spectrum was generated using FID-A to visualize the effect of the observed frequency drifts. The simulated spectrum was convolved with the frequency trace for the most extreme cases. Impacts of frequency drifts on NAA and GABA were also simulated as a function of linear drift. Data from the repeated protocol were compared with the corresponding first dataset using Pearson's and intraclass correlation coefficients (ICC).Results: Of the data collected from 99 scanners, 4 were excluded due to various reasons. Thus, data from 95 scanners were ultimately analyzed. For the first 5:20 min (64 transients), median (interquartile range) drift was 0.44 (1.29) Hz before fMRI and 0.83 (1.29) Hz after. This increased to 3.15 (4.02) Hz for the full 30 min (360 transients) run. Average drift rates were 0.29 Hz/min before fMRI and 0.43 Hz/min after. Paired t-tests indicated that drift increased after fMRI, as expected (p &lt; 0.05). Simulated spectra convolved with the frequency drift showed that the intensity of the NAA singlet was reduced by up to 26%, 44 % and 18% for GE, Philips and Siemens scanners after fMRI, respectively. ICCs indicated good agreement between datasets acquired on separate days. The single site long acquisition showed drift rate was reduced to 0.03 Hz/min approximately three hours after fMRI.Discussion: This study analyzed frequency drift data from 95 3T MRI scanners. Median levels of drift were relatively low (5-min average under 1 Hz), but the most extreme cases suffered from higher levels of drift. The extent of drift varied across scanners which both linear and nonlinear drifts were observed.</p

Track A Basic Science

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/138319/1/jia218438.pd

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Severe sepsis : epidemiology and sex-related differences in inflammatory markers

Background.  Sepsis is a syndrome associated with high mortality rates, substantial morbidity and high costs of care. The incidents of sepsis is reported to be high and controversy exists whether gender affect severity or outcome. Little is known about factors determining suscepti­bility for developing the syndrome and severity of the syndrome once developed. Early detection and adequate antibiotic administration are the mainstay of treatment and means to identify patients with particular high risk of adverse outcome are desirable. There are data to suggest that the course of sepsis and outcome from the syndrome may be influenced by inherited differences in the immunological response among humans Aims: Paper I: Assess incidence and outcome for ICU-treated sepsis patients in this region; Paper II: Assess if there are gender differences related to characteristics, aspects of treatment or out­come in sepsis in this region. Paper III: Assess the association of baseline levels of leptin and adiponectin and future sepsis event, and association of these adipokines in the cute phase and sepsis severity and outcome. Paper IV: Assess association of baseline levels of mannose-binding lectin (MBL) and future sepsis event, and MBL levels in the acute phase in relation to sepsis severity and outcome. Results. Paper I:  Overall ICU mortality rate was 25%, while the ICU mortality for patients with septic shock was 58% in this retrospective single university hospital cohort analysis. Cardio­vascular disease and diabetes were the most prevalent comorbidities among patients who died during hospital stay.  Paper II:  No gender-related differences in mortality or length of stay was found in this prospective single center observational study. Differences in aspects of treatment were related to differences in site of infection. Men had more often infections in skin and skin-structures, whereas women more often had abdominal infections. Early organ dysfunction asses­sed as SOFA score at admission was a stronger predictor for hospital mortality for women than for men. The discrepancy was related to the SOFA coagulation-sub score.  Paper III: In this nes­ted case-referent study hyperleptinemia at baseline predicted a first-ever sepsis event, even after adjustment for BMI and other cardiovascular risk factors. Hyperleptinemia in the acute sepsis phase was associated with reduced risk of in-hospital death in men, but associated with increased risk of in-hospital   death in women.  Paper IV: In the same matched cohort as in Paper III high baseline levels of MBL predicted a first ever sepsis event. High MBL levels in the acute phase or an increase from baseline to the acute phase associate with increased in-hospital death in women but not in men. Low MBL levels was not identified as a risk for acute sepsis or in-hospital death. Conclusions. Mortality from severe sepsis is high, equally affecting men and women. There are differences in patient characteristics and inflammatory markers, which associate with in-hospital mortality differentially in men and women. Aspects of gender should be mandatory, and genetic analysis are desired in future sepsis research

Segmentrapportering : Är den relevant ur en analytikers perspektiv?

Bakgrund och problemdiskussion: Med hjälp av segmentinformation kan analytiker bryta ned analysen av bolagen till en lägre nivå vilket borde resultera i säkrare analyser. Segmentinformationen är viktig för användarna av de finansiella rapporterna eftersom information om bolaget som helhet kan vara svår att utläsa. Vi har tagit del av tidigare forskning inom området,men vi har dock inte funnit någon forskning med samma syfte som vi har. Vi anser därför att segmentrapportering är viktigt och intressant att studera. Syfte: Vårt syfte med denna uppsats är att utreda hur, bolag noterade på Stockholmsbörsen i branschen sällanköpsvaror och tjänster, utformat sina segmentrapporter i deras årsredovisningar samt hur relevanta, tillförlitliga och jämförbara de är. Vidare kommer vi att studera vad analytiker anser om rapporterna gällande dessa egenskaper med fokus på hur relevanta de är. Metod: I denna uppsats använder vi oss av både kvalitativ och kvantitativ metod. Vi har genomfört tre besöksintervjuer med analytiker samt granskat 36 bolags segmentrapporter. Analytikerna har den specialistkunskap som krävs för att kunna svara på om relevansen är uppfylld för segmentrapporterna. Den kvantitativa metoden ger oss en annan synvinkel, den avspeglar hur bolagens segmentrapporter är utformade i verkligheten. Analys och slutsatser: För att analytikerna ska kunna framställa bra analyser så krävs det att informationen från segmentrapporterna är tillförlitlig och jämförbar och därigenom relevant. Om segmentrapporterna inte är relevanta leder detta till att analytikernas analyser blir felaktiga. Risken med detta är att läsarna av rapporterna baserar sina investeringar på felaktiga grunder vilket kan leda till ekonomiska förluster för dem. Respondenterna hade varierande åsikter gällande segmentrapporterna, men vi har funnit att segmentrapporterna i branschen generellt sett är tillförlitliga och jämförbara. Med detta som grund kan vi fastställa att de flesta av de studerade segmentrapporterna är relevanta. Vi vill dock poängtera att det finns en viss variation gällande hur relevanta rapporterna är. Nyckelord; segment, segmentrapportering, värdering, IAS 14, valuation, financial reporting,segment report(ing)

Förvaltningsfastigheter : Val av värderingsmetod vid värdering till verkligt värde

Bakgrund och problemdiskussion: Införandet av att svenska noterade företag måste tillämpa IFRS/IAS i sina årsredovisningar, har inneburit stora förändringar för berörda företags redovisning. En av dessa standarder är IAS 40 och dess huvudinnehåll behandlar hur ett företag ska värdera sina förvaltningsfastigheter. Det intressanta i dagens läge är att förvaltningsfastighetsföretagen får göra ett val av värderingsmetod vid värdering till verkligt värde. Syfte: Vårt syfte med denna uppsats är att kartlägga hur olika svenska förvaltningsfastighetsföretag, belägna i olika branscher och geografiska regioner, värderar sina förvaltningsfastigheter till verkligt värde efter införandet av IAS 40. Vi vill vidare analysera om det finns ett mönster vid fastighetsföretagens val av värderingsmetod. Metod: Vi har för avsikt att använda oss av en kvantitativ metod. Anledningen till vårt val av kvantitativ metod är att vi anser att vi kan få fram den mest relevanta och tillförlitliga informationen genom att studera våra valda företags årsredovisningar. Vi har valt att ta med hela populationen av svenska förvaltningsfastighetsföretag som tillämpar IAS 40. De sökord som har använts är; IAS 40, förvaltningsfastigheter, fastighetsföretag, värderingsmetod, värderingsmodell, fastighetsvärdering. Analys och slutsatser: Eftersom det är en stor del av vår population (19 av 21 företag) som använder kassaflödesmetoden kan vi inte se något samband gällande fastighetsföretagens storlek, bransch, geografisk lokalisering och val av värderingsmetod. De övriga två metoderna, ortsprismetoden och direktavkastningsmetoden, har vi kunnat se vissa mönster vid val av värderingsmetod. Till stor del stämmer vår teori, men inte helt. Vi har kommit fram till att i de fall ortsprismetoden och/eller en övrig värderingsmetod används tillsammans med kassaflödesmetoden, så bedömer vi att dessa främst har använts som ett komplement. Detta på grund av att informationen och tillvägagångssättet är begränsat i årsredovisningarna. Det ser vi som ett tecken på att tyngdpunkten i värderingen ligger i kassaflödesmetoden

Förvaltningsfastigheter : Val av värderingsmetod vid värdering till verkligt värde

Bakgrund och problemdiskussion: Införandet av att svenska noterade företag måste tillämpa IFRS/IAS i sina årsredovisningar, har inneburit stora förändringar för berörda företags redovisning. En av dessa standarder är IAS 40 och dess huvudinnehåll behandlar hur ett företag ska värdera sina förvaltningsfastigheter. Det intressanta i dagens läge är att förvaltningsfastighetsföretagen får göra ett val av värderingsmetod vid värdering till verkligt värde. Syfte: Vårt syfte med denna uppsats är att kartlägga hur olika svenska förvaltningsfastighetsföretag, belägna i olika branscher och geografiska regioner, värderar sina förvaltningsfastigheter till verkligt värde efter införandet av IAS 40. Vi vill vidare analysera om det finns ett mönster vid fastighetsföretagens val av värderingsmetod. Metod: Vi har för avsikt att använda oss av en kvantitativ metod. Anledningen till vårt val av kvantitativ metod är att vi anser att vi kan få fram den mest relevanta och tillförlitliga informationen genom att studera våra valda företags årsredovisningar. Vi har valt att ta med hela populationen av svenska förvaltningsfastighetsföretag som tillämpar IAS 40. De sökord som har använts är; IAS 40, förvaltningsfastigheter, fastighetsföretag, värderingsmetod, värderingsmodell, fastighetsvärdering. Analys och slutsatser: Eftersom det är en stor del av vår population (19 av 21 företag) som använder kassaflödesmetoden kan vi inte se något samband gällande fastighetsföretagens storlek, bransch, geografisk lokalisering och val av värderingsmetod. De övriga två metoderna, ortsprismetoden och direktavkastningsmetoden, har vi kunnat se vissa mönster vid val av värderingsmetod. Till stor del stämmer vår teori, men inte helt. Vi har kommit fram till att i de fall ortsprismetoden och/eller en övrig värderingsmetod används tillsammans med kassaflödesmetoden, så bedömer vi att dessa främst har använts som ett komplement. Detta på grund av att informationen och tillvägagångssättet är begränsat i årsredovisningarna. Det ser vi som ett tecken på att tyngdpunkten i värderingen ligger i kassaflödesmetoden