Detection of antigenic heterogeneity in feline coronavirus nucleocapsid in feline pyogranulomatous meningoencephalitis

A new monoclonal antibody (mAb), CCV2-2, was compared with the widely used FIPV3-70 mAb, both directed against canine coronavirus (CCoV), as a diagnostic and research tool. Western blot showed that both anti-CCoV mAbs only reacted with a protein of 50 kD, a weight consistent with the feline coronavirus (FCoV) viral nucleocapsid. A competitive inhibition enzyme-linked immunosorbent assay showed that the 2 recognized epitopes are distinct. Preincubation of CCV2-2 mAb with FCoV antigen suppressed the immunostaining. Formalin-fixed, paraffin-embedded sections from brains of 15 cats with the dry form of feline infectious peritonitis (FIP) were examined by immunohistochemistry. Immunohistochemistry was performed with both anti-CCoV mAbs, either on consecutive or on the same sections. A myeloid-histiocytic marker, MAC 387, was also used to identify FIP virus-infected cells. In all regions where MAC 387-positive cells were present, positive staining with the CCV2-2 mAb was systematically detected, except at some levels in 1 cat. In contrast, none or only a few cells were positive for the FIPV3-70 mAb. Double immunostaining showed macrophages that were immunopositive for either CCV2-2 alone or alternatively for CCV2-2 and FIPV3-70 mAbs. This reveals the coexistence of 2 cohorts of phagocytes whose FIP viral contents differed by the presence or absence of the FIPV3-70-recognized epitope. These findings provide evidence for antigenic heterogeneity in coronavirus nucleocapsid protein in FIP lesions, a result that is in line with molecular observations. In addition, we provide for the first time morphologic depiction of viral variants distribution in these lesions.     

Soil moisture, carbon and nitrogen dynamics following incorporation and surface application of labelled crop residues in soil columns

One way to increase the amount of carbon sequestered in agricultural land is to convert conventional tillage into no‐tillage systems. This greatly affects the location of crop residues in soil. To investigate the impact of the location of residues on soil physical and biological properties and how the interactions between those properties influence the fate of carbon and nitrogen in soil, we did a laboratory experiment with repacked soil in columns. Doubly labelled ¹³C¹⁵N oilseed rape residues were incorporated in the 0–10 cm layer or left on the soil surface. The columns were incubated for 9 weeks at 20°C and were submitted to three cycles of drying and wetting, each of them induced by a rain simulator. The location of the residues affected the water dynamics and the distribution of C and N in the soil, which in turn influenced microbial activity and the decomposition rate of the added residues. After 9 weeks of’incubation, 18.4 ± 1.5% of the surface applied residue‐C and 54.7 ± 1.3% of the incorporated residue‐C was mineralized. We observed a nitrate accumulation of 10.7 mg N kg^?1 with residues at the soil surface, 3.6 mg N kg^?1 with incorporated residues and 6.3 mg N kg^?1 without addition of fresh organic matter, which entailed net N mineralization in soil under mulch and immobilization of N with residue incorporation compared with the control soil. We concluded that application of oilseed rape residues at the soil surface increased the storage of fresh organic C in soil in the short term, compared with the incorporation treatment, but increased the risk of nitrate leaching.     

The Solar Acoustic Spectrum and Eigenmode Parameters

The Global Oscillation Network Group (GONG) project estimates the frequencies, amplitudes, and linewidths of more than 250,000 acoustic resonances of the sun from data sets lasting 36 days. The frequency resolution of a single data set is 0.321 microhertz. For frequencies averaged over the azimuthal order m, the median formal error is 0.044 microhertz, and the associated median fractional error is 1.6 x 10(-5). For a 3-year data set, the fractional error is expected to be 3 x 10(-6). The GONG m-averaged frequency measurements differ from other helioseismic data sets by 0.03 to 0.08 microhertz. The differences arise from a combination of systematic errors, random errors, and possible changes in solar structure.     

Fullerene pipes

Single-wall fullerene nanotubes were converted from nearly endless, highly tangled ropes into short, open-ended pipes that behave as individual macromolecules. Raw nanotube material was purified in large batches, and the ropes were cut into 100- to 300-nanometer lengths. The resulting pieces formed a stable colloidal suspension in water with the help of surfactants. These suspensions permit a variety of manipulations, such as sorting by length, derivatization, and tethering to gold surfaces.     

Analytical study of azadirachtin and 3-tigloylazadirachtol residues in foliage and phloem of hardwood tree species by liquid chromatography-electrospray mass spectrometry

A rapid and sensitive LC-ESI-MS method has been developed and validated for the quantitation of azadirachtin and 3-tigloylazadirachtol in deciduous tree matrices. The method involves automated extraction and simultaneous cleanup using an accelerated solvent technique with the matrix dispersed in solid phase over a layer of primary-secondary amine silica. The limits of quantification were 0.02 mg/kg for all matrices with the exception of Norway maple foliage (0.05 mg/kg). Validation at three levels (0.02, 0.1, and 1 mg/kg), demonstrated satisfactory recoveries (71-103%) with relative standard deviation <20%. Two in-source fragment ions were used for confirmation at levels above 0.1 mg/kg. Over a period of several months, quality control analyses showed the technique to be robust and effective in tracking the fate of these natural botanical insecticides following systemic injection into various tree species for control of invasive insect pest species such as the emerald ash borer and Asian longhorned beetle.     

Continuous versus intermittent thermodilution for cardiac output measurement during alveolar recruitment manoeuvres in sheep

ObjectiveTo evaluate interchangeability of a thermodilution based STAT mode continuous cardiac output (CCO) measurement method with bolus thermodilution (BTD).Study designRandomized crossover study.AnimalsTen 9 month old healthy male sheep.MethodsEach sheep was anaesthetized twice for laparoscopy. On one occasion mechanical ventilation was used immediately after anaesthetic induction (IPPV treatment) and on the other occasion the start of IPPV was delayed and two periods of alveolar recruitment manoeuvres were also performed (RM treatment). Cardiac output (CO) was measured simultaneously with both CCO and BTD at 6 time points. Data were analysed using difference versus mean plots. A priori limits of acceptance were set at ±30% of the mean of every paired measurement. If <5% of the data fell outside of these limits (Chi-square test, p < 0.05) the interchangeability of methods was accepted. Proportions of data outside of these limits were also compared between treatments (Fisher's test, p < 0.05). Cardiac output data from each treatment and measurement method were also analyzed separately with one-factorial anova and Bonferroni test (p < 0.05).ResultsA total of 119 measurements were obtained. Cardiac output ranged from 1.9 to 10.4 L minute^?1 (CCO) and from 1.1 to 9.8 L minute^?1 (BTD). The bias and limits of agreement were 0.5 ± 1.9 L minute^?1. More than 5% of all data fell outside of the limits of acceptance (24/119), and a larger proportion fell outside of these limits in the RM (20/59) compared to the IPPV treatment (4/60). The Bonferroni test detected significant decreases of CO over time in both treatments when measured with BTD but not with CCO.Conclusions and clinical relevanceThe STAT mode CCO method is not interchangeable with BTD during acute haemodynamic changes caused by recruitment manoeuvres, thus the results of STAT mode CCO should be interpreted with caution because decreases in CO may not be detected.     

Effect of Ram Age on Structural and Functional Competence of Frozen–Thawed Spermatozoa in Dairy Sheep

The objectives of this study were to investigate the influence of ram age on structural and functional competence of frozen–thawed spermatozoa and to test the hypothesis that increasing number of sperm bound to the zona pellucida in vitro was associated with decreasing in vivo fertility of frozen semen. Rams were allocated into two groups. Each group consisted of five rams aged either 1–2 years (young) or 4–5 years (mature). Three successive ejaculates were collected from each ram using an artificial vagina. Only ejaculates of ≥ 2.5 × 10⁹ sperm/ml and 80% sperm progressive motility were pooled per ram, diluted with Bioxcell^® medium and frozen in 0.25 ml straws. The end points of post‐thawing semen evaluation were computer‐assisted cell motility analysis, sperm capacitation (chlortetracycline assay), simultaneous assessment of plasma membrane integrity, mitochondrial membrane potential and condensation status of nucleus, per‐cell analysis of lipid peroxidation using C11‐BODIPY^581/591, sperm‐hemizona binding (HZB) ability and sperm fertility after laparoscopic insemination of ewes (n = 114) in the progestagen‐synchronized oestrus. The results showed that mature rams had significantly lower values of sperm hyperactivated motility and peroxidized sperm, higher percentages of live non‐capacitated sperm and sperm cells with intact plasma membrane, functional mitochondria and condensed chromatin, as well as, greater lambing rate and ewe prolificacy. Sperm HZB binding ability was higher (p < 0.05) for young than for mature rams. Significant correlations were found between number of spermatozoa bound to the zona pellucida and semen fertility (r = ?0.63 to ?0.71). In conclusion, mature rams have better semen quality and in vivo fertility than young rams. Cryocapacitation can be involved in decreasing ram semen fertility as evidenced by the high number of spermatozoa bound to the zona pellucida in vitro.     

Cardiorespiratory effects of hemic versus nonhemic prime during and immediately following mitral valve replacement in dogs.

In order to limit the hemodilution effect during cardiopulmonary bypass (CPB) in low weight animal patients, blood is often used as a component of the prime solution. This study was undertaken to evaluate the effects of the addition of blood to the prime solution on the hemodynamic and respiratory parameters during and following mitral valve replacement in dogs. Ten dogs were randomly assigned to receive either a hemic (HP), 75% blood component, or a nonhemic prime (NP) solution. The hemodilution was 5 +/- 4% and 25 +/- 10% for the HP and NP groups, respectively. Cardiopulmonary measurements were taken 20 minutes before initiating CPB, during CPB, and 20 min after termination. The hematocrit level, the hemoglobin concentration, and the arterial oxygen content were significantly lower in the NP group during and following CPB. However, the systemic oxygen transport index was not significantly different between the NP group (355 +/- 87 mL/min/m2) and the HP group (546 +/- 155 mL/min/m2) following CPB. Our study indicates that, in normal dogs undergoing hemodilution from a nonhemic prime solution, the cardiovascular function is able to maintain the systemic oxygen transport in the period immediately following mitral valve replacement.