Use of albumin quotient and IgG index to differentiate blood- vs brain-derived proteins in the cerebrospinal fluid of cats with feline infectious peritonitis

BACKGROUND: Inflammation of the central nervous system (CNS) is a frequent condition in cats but etiology often remains unsolved. Routine cerebrospinal fluid (CSF) analysis can be extended through the calculation of the albumin quotient (Q(alb)), a marker of the integrity of the blood-brain barrier (BBB), and IgG index, an estimate of intrathecal IgG synthesis. OBJECTIVES: The purpose of this study was to validate nephelometric methods for CSF protein analysis, and to use the Q(alb) and IgG index to discriminate blood- and brain-derived immunoglobulin fractions in cats with feline infectious peritonitis (FIP). METHODS: Cats presented to our clinic between 2001 and 2005 were included in the study based on clinical and laboratory data and histopathologic findings at necropsy. Cats were grouped as having nonneurologic disease (controls; n=37), brain tumors (n=8), FIP involving the CNS (n=12), and extraneural FIP (n=12). CSF-total protein (TP) was measured and albumin and IgG concentrations were measured in paired CSF/serum samples; Q(alb) and IgG index were calculated. Intraassay and interassay precision of the nephelometric assays were determined using pooled samples. RESULTS: Coefficients of variation for the nephelometric assays ranged from 2.7% to 7.2%. In control cats, CSF-TP concentration ranged from 0.06 to 0.36 g/L, Q(alb) ranged from 0.6 to 5.7 x 10(-3), and IgG index ranged from 0.3 to 0.6. Q(alb) and IgG index were significantly higher in cats with brain tumors and cats with CNS-FIP compared with other groups. Compared with control cats, pleocytosis was evident in 8 of 12 (67%) cats and CSF-TP was increased in 3 of 12 (25%) cats with CNS-FIP. CONCLUSION: Nephelometry is a reliable method for measurement of CSF protein, albumin, and IgG in cats. The Q(alb) and IgG index did not identify a CSF protein pattern specific for BBB dysfunction or intrathecal IgG synthesis in cats with CNS-FIP.     

Ultrasonographic appearance of the craniocervical junction in normal brachycephalic dogs and dogs with caudal occipital (Chiari-like) malformation

Ultrasonographic evaluation of the spine, especially of the spinal cord, has been rarely reported in dogs. The atlanto-occipital junction provides a small acoustic window through which examination of the craniocervical transition can be performed. Normal sonographic findings of this region in 10 normal brachycephalic dogs are presented and compared with sonographic findings from 25 Cavalier King Charles Spaniels with the caudal occipital malformation syndrome. Sonographic findings were compared with magnetic resonance imaging findings to determine the extent of cerebellar herniation and syringohydromyelia. Cerebellar displacement into the foramen magnum was clearly identified sonographically; however, syringohydromyelia was not discernable due to bone overlay.     

Iron oxide‐humic acid coprecipitates as iron source for cucumber plants

In soil, iron (Fe) solubility depends on complex interactions between Fe minerals and organic matter, but very little is known about plant availability of Fe present in Fe oxides associated with humic substances. For this purpose, this study investigates the effect of Fe mineral crystallinity in the presence of humic acids (HA) on Fe availability to plants. Four Fe–HA mineral coprecipitates were prepared, either in the presence or absence of oxygen, i.e., two goethite (G)‐HA samples containing large amounts of Fe as nanocrystalline goethite and ferrihydrite mixed phases, and two magnetite (M)‐HA samples containing crystalline magnetite. Bioavailability studies were conducted in hydroponic systems on cucumber plants (Cucumis sativus L.) grown under Fe deficient conditions and supplied with the Fe–HA coprecipitates containing goethite or magnetite. Results showed that plants grown in the presence of Fe–HA coprecipitates exhibited a complete recovery from Fe deficiency, albeit less efficiently than plants resupplied with Fe‐chelate fertilizer used as control (Fe‐diethylene triamine penta acetic acid, Fe‐DTPA). However, the supply with either G‐ or M–HA coprecipitates produced different effects on plants: G–HA‐treated plants showed a higher Fe content in leaves, while M–HA‐treated plants displayed a higher leaf biomass and SPAD (Soil–Plant Analysis Development) index recovery, as compared to Fe‐DTPA. The distribution of macronutrients in the leaves, as imaged by micro X‐ray fluorescence (µXRF) spectroscopy, was different in G–HA and M–HA‐treated plants. In particular, plants supplied with the poorly crystalline G–HA coprecipitate with a lower Fe/HA ratio showed features more similar to those of fully recovered plants (supplied with Fe‐DTPA). These results highlight the importance of mineral crystallinity of Fe–HA coprecipitates on Fe bioavailability and Fe uptake in hydroponic experiments. In addition, the present data demonstrate that cucumber plants can efficiently mobilize Fe, even from goethite and ferrihydrite mixed phases and magnetite, which are usually considered unavailable for plant nutrition.     

Influence of High Molecular Weight Glutenins on Viscoelastic Properties of Intact Wheat Kernel and Relation to Functional Properties of Wheat Dough

High and low molecular weight glutenin subunits (HMW‐GS and LMW‐GS, respectively) are the main factors determining the viscoelastic properties of wheat dough. The mechanical and viscoelastic properties of 29 samples of wheat kernels differing in HMW‐GS were evaluated with load‐compression tests. Samples were grouped by genotypes differing in HMW‐GS composition (allelic variants: Glu‐A1: null, 1, 2*; Glu‐B1: 7, 7+8, 7+9, 13+16, and 17+18; Glu‐D1: 5+10, 2+12). Groups representing Glu‐A1 1 and 2*; Glu‐B1 7, 7+9 and 17+18; and Glu‐D1 5+10 generally possessed hard grain and showed the largest kernel elasticity values, while those representing subunits Glu‐A1 null; Glu‐B1 7+8; and Glu‐D1 2+12 had soft kernels and showed lower elastic work values. Genotypes possessing HMW‐GS 1, 17+18 and 5+10 gave large SDS‐sedimentation values and better dough viscoelastic properties than those with allelels: null, 7+8, and 2+12. Kernel hardness showed significant correlation with the dough‐strength‐related parameters: SDS‐sedimentation; dough mixing time; and the alveographic parameters, W and P. There was a negative correlation between kernel plastic work and dough mixing time and the dough tenacity/extensibility parameters, P/L. The significant relationship between sedimentation tests and kernel elastic work seems to indicate that elastic work is related to genotype (protein composition). The general tendency was that higher values in kernel elastic work and size corresponded to better dough rheological quality. Mechanical properties of the kernel were significantly related to the elastic behavior measured in a single wheat kernel. The use of the compression test on individual kernels is easy, rapid and nondestructive and therefore seems to show potential use as a rapid tool in breeding to improve wheat quality.     

Distribution and composition of phytosterols and steryl ferulates in wheat grain and bran fractions

Phytosterols and steryl ferulates are bioactive compounds accumulating in the bran and germ of wheat. However, little is known regarding their localisation and composition in the bran layers of the kernel. The aim of this study was to determine the distribution of phytosterols and steryl ferulates in the wheat grain and in the different layers of bran. The wheat fractions, produced by conventional debranning, aleurone separation and a novel electrostatic process, were analysed for phytosterol contents using GC–FID and for steryl ferulate contents using HPLC–UV. The compounds were identified by GC– and LC–MS. Phytosterols and steryl ferulates were concentrated in the bran layers. The steryl ferulates were accumulated in the intermediate layers, whereas the phytosterols were more evenly distributed in the intermediate layers and aleurone cell contents. The phytosterol composition varied within the wheat kernel, while the steryl ferulate composition was similar in different fractions. Sitosterol and campestanyl ferulate were the main compounds. The highest levels of phytosterols (up to 2117 μg/g) and steryl ferulates (up to 703 μg/g) were found in the pearling, aleurone and certain bran fractions. The phytosterol-rich fractions could be utilised in cereal foods to enhance the intake of health-promoting compounds from natural sources.     

Proposal for field sampling of plants and processing in the lab for environmental metabolic fingerprinting

Background  

Samples for plant metabolic fingerprinting are prepared generally by metabolism quenching, grinding of plant material and extraction of metabolites in solvents. Further concentration and derivatisation steps follow in dependence of the sample nature and the available analytical platform. For plant material sampled in the field, several methods are not applicable, such as, e.g., collection in liquid nitrogen. Therefore, a protocol was established for sample pre-treatment, grinding, extraction and storage, which can be used for analysis of field-collected plant material, which is further processed in the laboratory. Ribwort plantain (Plantago lanceolata L., Plantaginaceae) was used as model plant. The quality criteria for method suitability were high reproducibility, extraction efficiency and handling comfort of each subsequent processing step.     

Identification and apoptotic potential of T-2 toxin metabolites in human cells

The mycotoxin T-2 toxin, produced by various Fusarium species, is a widespread contaminant of grain and grain products. Knowledge about its toxicity and metabolism in the human body is crucial for any risk assessment as T-2 toxin can be detected in processed and unprocessed food samples. Cell culture studies using cells of human origin represent a potent model system to study the metabolic fate of T-2 toxin as well as the cytotoxicity in vitro. In this study the metabolism of T-2 toxin was analyzed in a cell line derived from human colon carcinoma cells (HT-29) and primary human renal proximal tubule epithelial cells (RPTEC) using high-performance liquid chromatography coupled with Fourier transformation mass spectrometry (HPLC-FTMS). Both cell types metabolized T-2 toxin to a variety of compounds. Furthermore, cell cycle analysis in RPTEC proved the apoptotic effect of T-2 toxin and its metabolites HT-2 toxin and neosolaniol in micromolar concentrations.