Screening 34 Peanut Introductions for Allergen Content Using ELISA

Peanut is one of the most allergenic foods and reports of accidental ingestion of peanuts in unsuspected food are increasing. No information is available on the allergen content of peanut germplasm grown commercially and used in the food and confectionery industry. The objectives of this study were: (1) to determine the allergen contents of 34 peanut introductions (PI); and (2) to identify naturally occurring allergen-free and/or low or hypoallergenic peanuts germplasm. A basic ELISA protocol was utilized to detect the presence of antigens in the peanut lines using a pool of human sera from patients with documented history of peanut allergy. Two naturally occurring low, or hypo-allergenic germplasm were identified as PI 261942 and PI 338386. Both are Valencia market types with total allergen content significantly lower (P ≤0.05) than that of PI 119880 (0.550), PI 119876 (0.415) and PI 118991 (0.410) three Valencia market types and PI 262111 (0.485), a Virginia market type. No allergen-free PI was found. Allergen content of peanut lines from Bolivia and Paraguay were significantly (P ≤0.05) different to those from Venezuela. No significant difference was observed in the allergen content of the four market types.     

Isoflurane disrupts anterio-posterior phase synchronization of flash-induced field potentials in the rat

Consciousness presumes a set of integrated functions such as sensory processing, attention, and interpretation, and may depend upon both local and long-range phase synchronization of neuronal activity in cerebral cortex. Here we investigated whether volatile anesthetic isoflurane at concentrations that produce loss of consciousness (LOC) disrupts long-range anterio-posterior and local anterior synchronization of neuronal activity in the rat. In six rats, deep electrodes were chronically implanted in the primary visual cortex (V1) and in two areas of the motor cortex (M1 and M2) for recording of intracortical event-related potentials (ERP). Thirty discrete flashes were presented at random interstimulus intervals of 15–45 s, and ERPs were recorded at stepwise increasing isoflurane concentrations of 0–1.1%. Neuronal synchronization was estimated using wavelet coherence computed from the ERP data band-pass filtered at 5–50 Hz. We found that (1) in the waking state, long-range anterio-posterior coherence in 5–25 Hz and 25–50 Hz frequency bands was significantly higher than local anterior coherence; (2) anterio-posterior coherence in both 5–25 Hz and 26–50 Hz bands was significantly reduced by isoflurane in a concentration-dependent manner; (3) local anterior coherence was not affected by isoflurane at any of the concentrations studied. These findings suggest that a disruption of long-range anterio-posterior rather than local anterior synchronization of neuronal activity precedes the anesthetic-induced loss of consciousness.     

Lymph Node Mesenchymal and Endothelial Stromal Cells Cooperate via the RANK-RANKL Cytokine Axis to Shape the Sinusoidal Macrophage Niche

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Ca2+ permeability of the plasma membrane induced by rotavirus infection in cultured cells is inhibited by tunicamycin and brefeldin A

Rotavirus infection of cultured cells induces a progressive increase in plasma membrane permeability to Ca2+. The viral product responsible for this effect is not known. We have used tunicamycin and brefeldin A to prevent glycosylation and membrane traffic and study the involvement of viral glycoproteins, NSP4 and/or VP7, in rotavirus-infected HT29 and MA104 cells. In infected cells, we observed an increase of plasma membrane Ca2+ permeability and a progressive depletion of agonist-releasable ER pools measured with fura 2 and an enhancement of total Ca2+ content measured as 45Ca2+ uptake. Tunicamycin inhibited the increase in membrane Ca2+ permeability, induced a depletion of agonist-releasable and 45Ca2+-sequestered pools. Brefeldin A inhibited the increase of Ca2+ permeability and the increase in 45Ca2+ uptake induced by infection. We propose that the glycosylated viral product NSP4 (and/or VP7) travels to the plasma membrane to form a Ca2+ channel and hence elevate Ca2+ permeability.     

Comparison of hepatitis C viral loads in patients with or without coinfection with different genotypes

Hepatitis C virus genotyping was assessed for 257 chronic hepatitis C patients with viral loads above 1,000 IU/ml. Twelve patients were coinfected with more than one genotype. Their median viral loads did not differ significantly from those observed for monoinfected patients, which in turn did not vary significantly among different genotypes.     

Kinetic analysis of interactions between bispecific monoclonal antibodies and immobilized antigens using a resonant mirror biosensor

A resonant mirror biosensor (IAsys) protocol is described for the comparative kinetic analysis of the ability of monoclonal antibodies (Mabs) and bispecific antibodies (Babs) to bind immobilized antigens. The protocol has been optimized and validated using the panel of affinity-purified antibodies, including two parental Mabs, one specific to human immunoglobulin G (hIgG) and another specific to horseradish peroxidase (HRP), and a Bab derived thereof by cell fusion (anti-hIgG/HRP Bab). The real-time kinetic analysis of antigen–antibody interactions using this protocol allows to demonstrate the differences in the avidity of bivalently binding Mabs and monovalent Babs. As shown in our previous study [J. Immunol. Methods 261 (2002) 103], the observed equilibrium association constants (K_ass) determined by IAsys using this protocol yield figures almost overlapping with those obtained by solid-phase radioimmunoassay (RIA). The described protocol is suited for the investigation of the effects of valency on the binding properties of antibodies. It also may be applied for the selection of Mabs and Babs with desired features, for different fields of application.     

Identifying recent HIV infections using the avidity index and an automated enzyme immunoassay

We evaluated a procedure for identifying recent HIV infections, using sequential serum samples from 47 HIV-positive persons for whom the seroconversion date could be accurately estimated. Each serum sample was divided into two aliquots: one diluted with phosphate-buffered saline and the other diluted with 1 M guanidine. We assayed the aliquots with the automated AxSYM HIV1/2gO test (Abbott Diagnostics Division), without modifying the manufacturer's protocol. We then calculated the avidity index (AI): the ratio of the sample/cutoff value for the guanidine aliquot to that of the phosphate-buffered saline aliquot. We analyzed 216 serum samples: 34 samples were collected within 6 months of seroconversion (recent seroconversions), and 182 were collected after 6 months. The mean AIs, by time from seroconversion, were 0.68 +/- 0.16 (within 6 months) and 0.98 +/- 0.10 (after 6 months) (P < 0.0001). AI of <0.90 correctly identified 88.2% of recent infections but misclassified as recent infections 13.2% of serum samples collected afterward. The probability of an infection being classified as recent and having AI of > or = 0.90 would be 0.7% in a population with 5% recent infections. AI can identify with a certain degree of accuracy recent HIV infections, and being a quantitative index, it provides different levels of sensitivity and specificity, depending on the selected cutoff value. The standard assay procedure is not modified. This test is simple and inexpensive and could be used for surveillance, decision-making in treatment, and prevention.     

Memory for object and object-location after lesions to the ventromedial prefrontal cortex in humans

The aim of the present study was to investigate the effect of small unilateral lesions to the ventromedial portion of the prefrontal cortex on two memory functions: memory for objects and memory for object locations. Patients, who had undergone surgery of the anterior communicating artery aneurysm, and normal control subjects, participated in the study. The patients were subdivided into two groups: with and without unilateral resection of the gyrus rectus. Subjects were presented with two memory tests, that required remembering either simultaneously presented visual stimuli (object memory test; OMT) or locations of the stimuli (location memory test; LMT). In the OMT, patients with resection of the gyrus rectus were impaired in comparison to patients without resection and normal control subjects. In the LMT, the three groups did not differ from each other. Our results suggest that the ventromedial prefrontal cortex is specifically involved in memory for objects.