RecNcMIC3-1-R is a microneme- and rhoptry-based chimeric antigen that protects against acute neosporosis and limits cerebral parasite load in the mouse model for Neospora caninum infection

In order to achieve host cell entry, the apicomplexan parasite Neospora caninum relies on the contents of distinct organelles, named micronemes, rhoptries and dense granules, which are secreted at defined timepoints during and after host cell entry. It was shown previously that a vaccine composed of a mixture of three recombinant antigens, corresponding to the two microneme antigens NcMIC1 and NcMIC3 and the rhoptry protein NcROP2, prevented disease and limited cerebral infection and transplacental transmission in mice. In this study, we selected predicted immunogenic domains of each of these proteins and created four different chimeric antigens, with the respective domains incorporated into these chimers in different orders. Following vaccination, mice were challenged intraperitoneally with 2 × 10(6)N. caninum tachzyoites and were then carefully monitored for clinical symptoms during 4 weeks post-infection. Of the four chimeric antigens, only recNcMIC3-1-R provided complete protection against disease with 100% survivors, compared to 40-80% of survivors in the other groups. Serology did not show any clear differences in total IgG, IgG1 and IgG2a levels between the different treatment groups. Vaccination with all four chimeric variants generated an IL-4 biased cytokine expression, which then shifted to an IFN-γ-dominated response following experimental infection. Sera of recNcMIC3-1-R vaccinated mice reacted with each individual recombinant antigen, as well as with three distinct bands in Neospora extracts with similar Mr as NcMIC1, NcMIC3 and NcROP2, and exhibited distinct apical labeling in tachyzoites. These results suggest that recNcMIC3-1-R is an interesting chimeric vaccine candidate and should be followed up in subsequent studies in a fetal infection model.

S-0 and S-1 State Structure, Methyl Torsional Barrier Heights, and Fast Intersystem Crossing Dynamics of 5-Methyl-2-hydroxypyrimidine

We report the analysis of the SI So rotational band contours of jet-cooled 5-methyl-2-hydroxypyrimidine (5M2HP), the enol form of deoxythymine. Unlike thymine, which exhibits a structureless spectrum, the vibronic spectrum of 5M2HP is well structured, allowing us to determine the rotational constants and the methyl group torsional barriers in the S-0 and S-1 states. The 0(0)(0), 6a(0)(1), 6b(0)(1), and 14(0)(1) band contours were measured at 900 MHz (0.03 cm(-1)) resolution using mass-specific two-color resonant two-photon ionization (2C-R2PI) spectroscopy. All four bands are polarized perpendicular to the pyrimidine plane (>90% c type), identifying the S-1 <- S-0 excitation of 5M2HP as a 1n pi* transition. All contours exhibit two methyl rotor subbands that arise from the lowest 5-methyl torsional states 0A '' and 1E ''. The S-0 and S-1 state torsional barriers were extracted from fits to the torsional subbands. The 3-fold barriers are V-3 '' = 13 cm(-1) and V3' = SI cm(-1); the 6-fold barrier contributions V-6 '' and V-6' are in the range of 2-3 cm(-1) and are positive in both states. The changes of A, B, and C rotational constants upon S-1 <- S-0 excitation were extracted from the contours and reflect an "anti-quinoidal" distortion. The 0(0)(0) contour can only be simulated if a 3 GHz Lorentzian line shape is included, which implies that the S-1(1n pi*) lifetime is similar to 55 ps. For the 6a(0)(1) and 6b(0)(1) bands, the Lorentzian component increases to 5.5 GHz, reflecting a lifetime decrease to similar to 30 ps. The short lifetimes are consistent with the absence of fluorescence from the 1n pi* state. Combining these measurements with the previous observation of efficient intersystem crossing (ISC) from the Si state to a long-lived T-1((3)n pi*) state that lies similar to 2200 cm(-1) below [S. Lobsiger, S. et al. Phys. Chem. Chem. Phys. 2010, 12, 5032] implies that the broadening arises from fast intersystem crossing with k(ISC) approximate to 2 x 10(10) s(-1). In comparison to 5-methylpyrimidine, the ISC rate is enhanced by at least 10 000 by the additional hydroxy group in position 2.

Magnetic resonance (MR) cholangiography: quantitative and qualitative comparison of 3.0 Tesla with 1.5 Tesla

OBJECTIVES: To determine quantitative and qualitative image quality in patients undergoing magnetic resonance (MR) cholangiography at 3.0 Tesla (T) compared with 1.5 T. MATERIALS AND METHODS: Fifty patients (30 women; mean age, 51 years) underwent MR cholangiography at 1.5 T; another 50 patients (25 women; mean age 51 years) were scanned at 3.0 T. MR sequence protocol consisted of breath-hold single-slice rapid acquisition with relaxation enhancement (RARE) and a respiratory-triggered 3D turbo spin echo (3D TSE) sequence. Maximum intensity projections were generated from the 3D TSE datasets. Contrast-to-noise ratio (CNR) measurements between the common bile duct (CBD), left and right intrahepatic duct (LHD, RHD), and periductal tissue were performed. Three radiologists assessed qualitatively the visibility of the CBD, LHD, and RHD and the overall diagnostic quality. RESULTS: Mean gain in CNR at 3.0 T versus 1.5 T in all 3 locations ranged for the RARE sequence from 7.7% to 38.1% and for the 3D TSE from 0.5% to 26.1% (P > 0.05 for all differences). Qualitative analysis did not reveal any significant difference between the 2 field strengths (P > 0.05). CONCLUSIONS: MR cholangiography at 3.0 T shows a trend toward higher CNR without improving image quality significantly.

Characteristics, determinants, and clinical relevance of CD4 T cell recovery to <500 cells/microL in HIV type 1-infected individuals receiving potent antiretroviral therapy

BACKGROUND: The CD4 T cell count recovery in human immunodeficiency virus type 1 (HIV-1)-infected individuals receiving potent antiretroviral therapy (ART) shows high variability. We studied the determinants and the clinical relevance of incomplete CD4 T cell restoration. METHODS: Longitudinal CD4 T cell count was analyzed in 293 participants of the Swiss HIV Cohort Study who had had a plasma HIV-1 RNA load <1000 copies/mL for > or =5 years. CD4 T cell recovery was stratified by CD4 T cell count 5 years after initiation of ART (> or =500 cells/microL was defined as a complete response, and <500 cells/microL was defined as an incomplete response). Determinants of incomplete responses and clinical events were evaluated using logistic regression and survival analyses. RESULTS: The median CD4 T cell count increased from 180 cells/microL at baseline to 576 cells/microL 5 years after ART initiation. A total of 35.8% of patients were incomplete responders, of whom 47.6% reached a CD4 T cell plateau <500 cells/microL. Centers for Disease Control and Prevention HIV-1 disease category B and/or C events occurred in 21% of incomplete responders and in 14.4% of complete responders (P>.05). Older age (adjusted odds ratio [aOR], 1.71 per 10-year increase; 95% confidence interval [CI], 1.21-2.43), lower baseline CD4 T cell count (aOR, 0.37 per 100-cell increase; 95% CI, 0.28-0.49), and longer duration of HIV infection (aOR, 2.39 per 10-year increase; 95% CI, 1.19-4.81) were significantly associated with a CD4 T cell count <500 cells/microL at 5 years. The median increases in CD4 T cell count after 3-6 months of ART were smaller in incomplete responders (P<.001) and predicted, in conjunction with baseline CD4 T cell count and age, incomplete response with 80% sensitivity and 72% specificity. CONCLUSION: Individuals with incomplete CD4 T cell recovery to <500 cells/microL had more advanced HIV-1 infection at baseline. CD4 T cell changes during the first 3-6 months of ART already reflect the capacity of the immune system to replenish depleted CD4 T lymphocytes.

Patient exposure and image quality of low-dose pulmonary computed tomography angiography: comparison of 100- and 80-kVp protocols

OBJECTIVE: Measures to reduce radiation exposure and injected iodine mass are becoming more important with the widespread and often repetitive use of pulmonary CT angiography (CTA) in patients with suspected pulmonary embolism. In this retrospective study, we analyzed the capability of 2 low-kilovoltage CTA-protocols to achieve these goals. MATERIALS AND METHODS: Ninety patients weighing less than 100 kg were examined by a pulmonary CTA protocol using either 100 kVp (group A) or 80 kVp (group B). Volume and flow rate of contrast medium were reduced in group B (75 mL at 3 mL/s) compared with group A (100 mL at 4 mL/s). Attenuation was measured in the central and peripheral pulmonary arteries, and the contrast-to-noise ratios (CNR) were calculated. Entrance skin dose was estimated by measuring the surface dose in an ovoid-cylindrical polymethyl methacrylate chest phantom with 2 various dimensions corresponding to the range of chest diameters in our patients. Quantitative image parameters, estimated effective dose, and skin dose in both groups were compared by the t test. Arterial enhancement, noise, and overall quality were independently assessed by 3 radiologists, and results were compared between the groups using nonparametric tests. RESULTS: Mean attenuation in the pulmonary arteries in group B (427.6 +/- 116 HU) was significantly higher than in group A (342.1 +/- 87.7 HU; P < 0.001), whereas CNR showed no difference (group A, 20.6 +/- 7.3 and group B, 22.2 +/- 7.1; P = 0.302). Effective dose was lower by more than 40% with 80 kVp (1.68 +/- 0.23 mSv) compared with 100 kVp (2.87 +/- 0.88 mSv) (P < 0.001). Surface dose was significantly lower at 80 kVp compared with 100 kVp at both phantom dimensions (2.75 vs. 3.22 mGy; P = 0.027 and 2.22 vs. 2.73 mGy; P = 0.005, respectively). Image quality did not differ significantly between the groups (P = 0.151). CONCLUSIONS: Using 80 kVp in pulmonary CTA permits reduced patient exposure by 40% and CM volume by 25% compared with 100 kVp without deterioration of image quality in patients weighing less than 100 kg.

Timing of initiation of antiretroviral therapy in AIDS-free HIV-1-infected patients: a collaborative analysis of 18 HIV cohort studies

BACKGROUND: The CD4 cell count at which combination antiretroviral therapy should be started is a central, unresolved issue in the care of HIV-1-infected patients. In the absence of randomised trials, we examined this question in prospective cohort studies. METHODS: We analysed data from 18 cohort studies of patients with HIV. Antiretroviral-naive patients from 15 of these studies were eligible for inclusion if they had started combination antiretroviral therapy (while AIDS-free, with a CD4 cell count less than 550 cells per microL, and with no history of injecting drug use) on or after Jan 1, 1998. We used data from patients followed up in seven of the cohorts in the era before the introduction of combination therapy (1989-95) to estimate distributions of lead times (from the first CD4 cell count measurement in an upper range to the upper threshold of a lower range) and unseen AIDS and death events (occurring before the upper threshold of a lower CD4 cell count range is reached) in the absence of treatment. These estimations were used to impute completed datasets in which lead times and unseen AIDS and death events were added to data for treated patients in deferred therapy groups. We compared the effect of deferred initiation of combination therapy with immediate initiation on rates of AIDS and death, and on death alone, in adjacent CD4 cell count ranges of width 100 cells per microL. FINDINGS: Data were obtained for 21 247 patients who were followed up during the era before the introduction of combination therapy and 24 444 patients who were followed up from the start of treatment. Deferring combination therapy until a CD4 cell count of 251-350 cells per microL was associated with higher rates of AIDS and death than starting therapy in the range 351-450 cells per microL (hazard ratio [HR] 1.28, 95% CI 1.04-1.57). The adverse effect of deferring treatment increased with decreasing CD4 cell count threshold. Deferred initiation of combination therapy was also associated with higher mortality rates, although effects on mortality were less marked than effects on AIDS and death (HR 1.13, 0.80-1.60, for deferred initiation of treatment at CD4 cell count 251-350 cells per microL compared with initiation at 351-450 cells per microL). INTERPRETATION: Our results suggest that 350 cells per microL should be the minimum threshold for initiation of antiretroviral therapy, and should help to guide physicians and patients in deciding when to start treatment.

Abnormalities of very low density lipoprotein apolipoprotein B-100 metabolism contribute to the dyslipidaemia of adult growth hormone deficiency

Increased cardiovascular mortality in adult growth hormone deficiency (GHD) may be, in part, explained by the dyslipidaemia associated with this condition. It is possible that abnormalities of very low density lipoprotein apolipoprotein B-100 (VLDL apoB) metabolism contribute to this dyslipidaemia. To test this hypothesis, we measured VLDL apoB kinetics in adult GH deficient patients (4 females, 3 males; age 50.1 +/- 4.7 yr (mean +/- SEM); BMI 28.2 +/- 1.1 kg/m2; total cholesterol (TC) 6.6 +/- 0.3 mmol/l; triglyceride (TG) 2.8 +/- 0.6 mmol/l; HDL cholesterol 1.1 +/- 0.1 mmol/l) and in control subjects (4 females, 3 male; age 47.0 +/- 4.7 yr; BMI 27.0 +/- 2.6 kg/m2; TC 5.0 +/- 0.4 mmol/l; TG 0.9 +/- 0.2 mmol/l; HDL cholesterol 1.4 +/- 0.1 mmol/l). [1-(13)C] leucine was administered by a primed (1 mg/kg), constant intravenous infusion (1 mg/kg/hr) and VLDL apoB enrichment with 13C leucine was determined using gas-chromatography mass-spectrometry. The GHD patients had a significantly higher hepatic secretion rate of VLDL apoB (15.5 +/- 1.8 mg/kg/day vs 9.4 +/- 0.6 mg/kg/day p = 0.007) and reduced catabolism ofVLDL apoB (metabolic clearance rate; 12.3 +/- 1.7 ml/min vs 24.3 +/- 4.8 ml/min p < 0.05) compared with control subjects. These findings suggest that GH is integrally involved in the regulation of VLDL apoB metabolism.

Follow-up of 100 dogs with acute diarrhoea in a primary care practice.

This study aimed to examine the aetiology of acute diarrhoea and the relapse rate in 100 client-owned dogs presented to a first-opinion clinic. History, physical examination, faecal testing and owner questionnaire data were collected at initial presentation (T0) and at either the time of relapse or at a recheck performed within 3 months. All dogs received treatment according to their clinical signs. Of 96 dogs that completed the study, 37 (38.5%) relapsed during the study period, 21 (21.9%) relapsed within 3 months, and 16 others (16.6%) at 3 months to 1 year after initial examination. Dogs that had undergone a change in housing location within 1 month prior to presentation and dogs <1 year old were significantly more likely to have positive parasitological analyses (P=0.02 and P=0.001, respectively). Pica was a risk factor for relapse (P=0.0002).

Comment on “The phase relation between atmospheric carbon dioxide and global temperature” Humlum et al. [Glob. Planet. Change 100: 51–69.]: Isotopes ignored

A recent study relying purely on statistical analysis of relatively short time series suggested substantial re-thinking of the traditional view about causality explaining the detected rising trend of atmospheric CO2 (atmCO2) concentrations. If these results are well-justified then they should surely compel a fundamental scientific shift in paradigms regarding both atmospheric greenhouse warming mechanism and global carbon cycle. However, the presented work suffers from serious logical deficiencies such as, 1) what could be the sink for fossil fuel CO2 emissions, if neither the atmosphere nor the ocean – as suggested by the authors – plays a role? 2) What is the alternative explanation for ocean acidification if the ocean is a net source of CO2 to the atmosphere? Probably the most provocative point of the commented study is that anthropogenic emissions have little influence on atmCO2 concentrations. The authors have obviously ignored the reconstructed and directly measured carbon isotopic trends of atmCO2 (both δ13C, and radiocarbon dilution) and the declining O2/N2 ratio, although these parameters provide solid evidence that fossil fuel combustion is the major source of atmCO2 increase throughout the Industrial Era.

Reconstruction and electrochemical oxidation of Au(110) surface in 0.1 M H2SO4

Variations of the surface structure and composition of the Au(110) electrode during the formation/lifting of the surface reconstruction and during the surface oxidation/reduction in 0.1 M aqueous sulfuric acid were studied by cyclic voltammetry, scanning tunneling microscopy and shell-isolated nanoparticle enhanced Raman spectroscopy. Annealing of the Au(110) electrode leads to a thermally-induced reconstruction formed by intermixed (1×3) and (1×2) phases. In a 0.1 M H2SO4 solution, the decrease of the potential of the atomically smooth Au(110)-(1×1) surface leads to the formation of a range of structures with increasing surface corrugation. The electrochemical oxidation of the Au(110) surface starts by the formation of anisotropic atomic rows of gold oxide. At higher potentials we observed a disordered structure of the surface gold oxide, similar to the one found for the Au(111) surface.

CO Oxidation on Pt(100): New Insights based on Combined Voltammetric, Microscopic and Spectroscopic Experiments

We present an experimental study of the CO electro-oxidation on Pt(100)-(1 × 1) electrodes employing electrochemical methods in combination with in situ scanning tunneling microscopy (STM) and shell-isolated nanoparticle enhanced Raman spectroscopy (SHINERS). We discussed the nature and stability of the active sites in the preignition region in the presence of dissolved CO (COb) and monitored substrate structure changes during the COb electro-oxidation process. We corroborated that the electro-oxidation kinetics is determined decisively by the history of CO adlayer formation. A new mechanism was proposed for Pt(100) electrode deactivation in the preignition region after excursion of electrode potential to COb ignition region. We believe that this mechanism takes place on Pt surfaces independently on their crystallographic orientation.