Depletion of CD8 alpha cells from tissues of Atlantic salmon during the early stages of infection with high or low virulent strains of infectious salmon anaemia virus (ISAV)

The virulence of an infectious salmon anaemia virus (ISAV) isolate is influenced by the response of the host's immune system to virus infection. Here we report the fate of immune responsive cells in head kidney, spleen and gills of Atlantic salmon during infection with high and low virulent strains of ISAV. A comparison of real-time PCR detection of virus and immunohistochemical detection of immune responsive cells revealed that peak viral load was coincident with both an elevated presence of MHC class I cells and a marked depletion of CD8 alpha cells. There was a larger CD8 alpha population in tissues from salmon infected with the low virulent strain compared with tissues from salmon infected with the high virulent strain at early stages of infection. These findings suggest a protective role for the CD8 alpha cell population in immune defences against ISAV.     

Annexin A2 as a differential diagnostic marker of hepatocellular tumors

While improved imaging techniques have made it possible to detect focal liver lesions smaller than 1 cm in diameter, differentiating benign lesions from hepatocellular carcinoma (HCC) still remains a challenge. To address this problem and obtain a definite diagnosis, needle core biopsies are often performed, leading to an increased need for supportive ancillary techniques in the histopathological assessment of highly differentiated hepatocellular tumors. Here we evaluate the diagnostic value of immunohistologically detected Annexin A2 (ANXA2) expression in highly differentiated liver tumors. ANXA2 is a calcium-dependent phospholipid-binding protein that has been linked to malignant transformation and HCC development. Our data show that adding sinusoidal ANXA2 expression to the already established marker panel (GPC3, GS, and HSP70) increases the accuracy for the detection of well-differentiated HCC (74% sensitivity, 100% specificity). In addition, in our series, the combinations ANXA2-GPC3 and ANXA2-GS performed better compared to the other established marker combinations. In conclusion, we suggest that adding ANXA2 to the established diagnostic marker panel increases the reliability and objectivity of HCC diagnosis in liver biopsies.     

Differential regulation of lipopolysaccharide and Gram-positive bacteria induced cytokine and chemokine production in macrophages by Galpha(i) proteins

Heterotrimeric G(i) proteins play a role in signalling activated by lipopolysaccharide (LPS), Staphylococcus aureus (SA) and group B streptococci (GBS), leading to production of inflammatory mediators. We hypothesized that genetic deletion of G(i) proteins would alter cytokine and chemokine production induced by LPS, SA and GBS stimulation. LPS-induced, heat-killed SA-induced and heat-killed GBS-induced cytokine and chemokine production in peritoneal macrophages from wild-type (WT), Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice were investigated. LPS induced production of tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-10 and interferon-gamma-inducible protein-10 (IP-10); SA induced TNF-alpha, and IL-1beta production; and GBS induced TNF-alpha, IL-6, IL-1beta, macrophage inflammatory protein-1alpha (MIP-1alpha) and keratinocyte chemoattract (KC) production were all decreased (P < 0.05) in Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice compared with WT mice. In contrast to the role of G(i) proteins as a positive regulator of mediators, LPS-induced production of MIP-1alpha and granulocyte-macrophage colony-stimulating factor (GM-CSF) were increased in macrophages from Galpha(i1/3) (-/-) mice, and SA-induced MIP-1alpha production was increased in both groups of Galpha(i) protein-depleted mice. LPS-induced production of KC and IL-1beta, SA-induced production of GM-CSF, KC and IP-10, and GBS-induced production of IL-10, GM-CSF and IP-10 were unchanged in macrophages from Galpha(i2) (-/-) or Galpha(i1/3) (-/-) mice compared with WT mice. These data suggest that G(i2) and G(i1/3) proteins are both involved and differentially regulate murine inflammatory cytokine and chemokine production in response to both LPS and Gram-positive microbial stimuli.     

Polysomnography findings in patients with restless legs syndrome and in healthy controls: a comparative observational study

STUDY OBJECTIVES: Sleep disturbances and their sequelae are the most common complaints of patients with restless legs syndrome (RLS). We compared polysomnography (PSG) findings in a large cohort of patients with idiopathic RLS and of healthy subjects. DESIGN: Comparative observational study. SETTING: University hospital sleep laboratory. PATIENTS: Age- and sex-matched patients with idiopathic but untreated RLS versus healthy controls. INTERVENTIONS: N/A RESULTS: Each group consisted of 29 females and 16 males. RLS subjects and controls were 47.4 +/- 10.9 and 47.3 +/- 10.5 years old, respectively. RLS severity was 24.0 +/- 6.2 points on the IRLS scale, indicating moderately severe RLS symptoms. We found strong multivariate group effects on PSG parameters (Wilks' lambda, P <0.001): RLS patients exhibited prolonged sleep onset latencies (according to the 10-min criterion but not to the one-epoch criterion), shorter total sleep time, lower sleep efficiency, higher arousal index, higher number of stage shifts, and longer REM sleep latency. During the sleep period time, percentage of wake and sleep stage 1 were increased, and sleep stage 2 and REM sleep were decreased in RLS patients. The PLMS indices and the sleep fragmentation index were markedly increased in the RLS group. CONCLUSIONS: We present the largest polysomnography study to date that compares patients with idiopathic RLS with age- and sex-matched healthy subjects. The findings demonstrate markedly fragmented sleep with deterioration of both NREM and REM sleep in RLS patients.     

Tracking the urinary excretion of high molar mass poly(vinyl alcohol)

The fate of poly(vinyl alcohol) (PVA) of weight average molar mass of 125,000 g/mol after administration into the peritoneum of rabbits has bean studied by various methods. PVA was spin-labeled with a nitroxide radical and then detected in urine using electron spin resonance (ESR) spectroscopy. Furthermore, unlabeled polymer was also administered to rabbits, then the urine was collected, dialyzed, precipitated, and the excretion of PVA was confirmed by size exclusion chromatography (SEC), FTIR spectroscopy, and (1)H NMR spectroscopy. ESR and SEC results show that, despite its relatively high molar mass, PVA is excreted through the kidneys without significant molar mass changes. Nevertheless, NMR and FTIR spectra show slight differences between the excreted and neat PVA. Possible causes of these discrepancies are discussed.     

Rare copy number variants in individuals at clinical high risk for psychosis: Enrichment of synaptic/brain‐related functional pathways

Schizophrenia is a complex and chronic neuropsychiatric disorder, with a heritability of around 60–80%. Large (>100 kb) rare (<1%) copy number variants (CNVs) occur more frequently in schizophrenia patients compared to controls. Currently, there are no studies reporting genome‐wide CNVs in clinical high risk for psychosis (CHR‐P) individuals. The aim of this study was to investigate the role of rare genome‐wide CNVs in 84 CHR‐P individuals and 124 presumably healthy controls. There were no significant differences in all rare CNV frequencies and sizes between CHR‐P individuals and controls. However, brain‐related CNVs and brain‐related deletions were significantly more frequent in CHR‐P individuals than controls. In CHR‐P individuals, significant associations were found between brain‐related CNV carriers and attenuated positive symptoms syndrome or cognitive disturbances (OR = 3.07, p = .0286). Brain‐related CNV carriers experienced significantly higher negative symptoms (p = .0047), higher depressive symptoms (p = .0175), and higher disturbances of self and surroundings (p = .0029) than noncarriers. Furthermore, enrichment analysis of genes was performed in the regions of rare CNVs using three independent methods, which confirmed significant clustering of predefined genes involved in synaptic/brain‐related functional pathways in CHR‐P individuals. These results suggest that rare CNVs might affect synaptic/brain‐related functional pathways in CHR‐P individuals.     

The Performance of Multileaf Collimators Evaluated by the Stripe Test

The performance of 3 multileaf collimator (MLC) systems (Varian Medical Systems, Elekta, and Siemens Medical Solutions) mounted on 7 different radiotherapy linear accelerators was investigated by a stripe test. The stripe test consisted of 8 adjacent multileaf segments of 2.5 × 40 cm², enclosed by all leaf pairs. With 6-MV photons, the segments were used to irradiate Agfa CR films. The optical density profile of the irradiated film in the travel direction of the MLC was used to estimate the short- and long-term leaf positioning reproducibility. The short-term reproducibility was found by analyzing 6 consecutive stripe tests. The long-term reproducibility was obtained by performing 3 to 5 stripe tests over 2 months. The short-term reproducibility was mainly within 0.3 mm for all systems. For the long-term reproducibility, the Varian and Elekta MLCs were within 0.4 to 0.5 mm, while the Siemens MLC showed a wider distribution, with values up to 1 mm for some leaf pairs. The inferior long-term reproducibility of the Siemens MLCs was mainly due to a decrease of the segment size with time. In conclusion, the stripe test is a useful method for evaluating MLC performance. Furthermore, the long-term reproducibility varied among the MLC systems investigated.     

Cinacalcet's effect on the pharmacokinetics of tacrolimus, cyclosporine and mycophenolate in renal transplant recipients.

BACKGROUND: The calcimimetic drug cinacalcet offers a novel therapeutic option to treat post-transplant hypercalcemia and hyperparathyroidism; however, the interaction with calcineurin inhibitors and mycophenolate has not been evaluated. METHODS: In the present study the effects of cinacalcet on the pharmacokinetics of cyclosporine A (CsA), tacrolimus (Tac) and mycophenolate were investigated in 14 renal transplant recipients with stable renal function (mean creatinine 126.4 +/- 45.3 micromol/L). The patients were treated with either CsA (n = 8) or Tac (n = 6) in combination with mycophenolate/azathioprine and steroids. Twelve-hour pharmacokinetic investigations to measure CsA and its six main metabolites, Tac and mycophenolate concentrations were performed before and after 1-week treatment with 30 mg cinacalcet once daily. RESULTS: Cinacalcet treatment induced a significant 14.3 +/- 12.1% decrease in Tac AUC(0-12) (P = 0.039). Tac C(max), T(max) and T(1/2) also tended to decrease. The pharmacokinetics of CsA and mycophenolate were not significantly affected by concomitant treatment with cinacalcet. However, the secondary CsA metabolite, AM19, showed a significant increase of 9.0 +/- 9.5% during cinacalcet treatment (P = 0.040). Renal function decreased significantly from 78 +/- 11 to 72 +/- 12 mL/min (P = 0.019) and correlated with the increased levels of metabolite AM19 in the CsA group. Renal function was unchanged in the Tac group. CONCLUSION: Cinacalcet treatment showed a moderate effect on the Tac, but not CsA or mycophenolate, pharmacokinetics after 1-week concomitant treatment. This interaction appears to have minor clinical relevance. However, it is advisable to monitor renal function in CsA-treated patients due to the observed decrease in renal function.     

Thrombospondins: multimodular proteins with angiostatic function]

The thrombospondins (TSPs) are a family of multimodular proteins that bind to the extracellular matrix with strong affinity. Of the five members of the TSP family, TSP1 and TSP2 are the only ones that inhibit endothelial cell migration in vitro and neoangiogenesis in vivo. This angiogenesis-inhibiting effect is mediated by interaction of a short sequence in type I modules with the membrane receptor CD36. TSP1 and TSP2 gene knockout experiments in mice showed increased blood vessel density in TSP2 -/- but no such alteration in TSP1 -/- animals. Loss of TSP1 gene expression was correlated with acquisition of an angiogenic phenotype in several models of human malignant tumors. Taken in concert, these findings suggest that TSP1 and, to a lesser extent, TSP2 may have therapeutic potential as angiogenesis-inhibiting factors.