Quantification of allergic and irritant patch test reactions using ImageJ.

BACKGROUND/AIMS: It is often difficult to differentiate between allergic and irritant patch test reactions by visual inspection. The purpose of this study was to test an image analysis-based method that differentiates between the two reactions by quantifying the degree of erythema at the patch test site. METHODS: A total of 172 Japanese patients were patch-tested with sodium lauryl sulfate (SLS) and nickel sulfate, followed by digital photography and visual evaluation of the patch test areas by dermatologists at 48 and 72 h. The digital images were converted to erythema index (EI) images by image processing, and changes in ΔEI (the difference in the EI between the patch test site and the adjacent normal skin) values were analyzed. RESULTS: The ΔEI was significantly increased at 72 h relative to that at 48 h for positive nickel sulfate reactions (P < 0.0001), while no significant difference in the ΔEI was found for SLS reactions. CONCLUSION: Using image analysis, allergic patch test reactions may be distinguished from irritant reactions by evaluating the change in the degree of erythema at 48 and 72 h.

Brain arachidonic acid incorporation and precursor pool specific activity during intravenous infusion of unesterified [3H]arachidonate in the anesthetized rat.

Brain fatty acid incorporation into phospholipids can be measured in vivo following intravenous injection of fatty acid tracer. However, to calculate a cerebral incorporation rate, knowledge is required of tracer specific activity in the final brain precursor pool. To determine this for one tracer, unesterified [3H]arachidonate was infused intravenously in pentobarbital-anesthetized rats to maintain constant plasma specific activity for 1-10 min. At the end of infusion, animals were killed by microwave irradiation and analyzed for tracer specific activity and concentration in brain phospholipid, neutral lipid, and lipid precursor, i.e., unesterified arachidonate and arachidonoyl-CoA, pools. Tracer specific activity in brain unesterified arachidonate and arachidonoyl-CoA rose quickly (t1/2 < 1 min) to steady-state values that averaged < 5% of plasma specific activity. Incorporation was rapid, as > 85% of brain tracer was present in phospholipids at 1 min of infusion. The results demonstrate that unesterified arachidonate is rapidly taken up and incorporated in brain but that brain phospholipid precursor pools fail to equilibrate with plasma in short experiments. Low brain precursor specific activity may result from (a) dilution of label with unlabeled arachidonate from alternate sources or (b) precursor pool compartmentalization. The results suggest that arachidonate turnover in brain phospholipids is more rapid than previously assumed.

trk immunoreactivity at neuronal dendrite and cell body.

Neurotrophins homologous to the nerve growth factor (NGF) bind the neurotrophic receptors of the trk gene family. Since the target tissues release these neurotrophic factors to the neuron, it has been believed that their trophic effects are mediated by the retrograde axonal transport. However it remains an open question whether the neurotrophins act through the autocrine or the paracrine mechanisms, since the protein-level expression of trk has not been studied so far. We have made polyclonal antibodies against the recombinant proteins of chicken trkC and rat trkB. These antibodies showed immunoreactivity at the dendrite and the cell body of neuron. This subcellular localization strongly suggests the autocrine or the paracrine mechanism of the neurotrophic factors. At the same time, our data provide basic knowledge to decide where to deliver these neurotrophic factors in the therapy of neurodegenerative diseases.

Identification of two molecular species of rat brain phosphatidylcholine that rapidly incorporate and turn over arachidonic acid in vivo.

In vivo rates of arachidonic acid incorporation and turnover were determined for molecular species of rat brain phosphatidylcholine (PtdCho) and phosphatidylinositol (PtdIns). [3H]Arachidonic acid was infused intravenously in pentobarbital-anesthetized rats at a programmed rate to maintain constant plasma specific activity for 2-10 min. At the end of infusion, animals were killed by microwave irradiation, and brain phospholipids were isolated, converted to diacylglycerobenzoates, and resolved as molecular species by reversed-phase HPLC. Most [3H] arachidonate (> 87%) was incorporated into PtdCho and PtdIns, with arachidonic acid at the sn-2 position and with oleic acid (18:1), palmitic acid (16:0), or stearic acid (18:0) at the sn-1 position. However, 10-15% of labeled brain PtdCho eluted in a small peak containing two molecular species with arachidonic acid at the sn-2 position and palmitoleic acid (16:1) or linoleic acid (18:2) at the sn-1 position. Analysis demonstrated that tracer was present in both the 16:1-20:4 and 18:2-20:4 PtdCho species at specific activities 10-40 times that of the other phospholipids. Based on the measured mass of arachidonate in each phospholipid molecular species, half-lives were calculated for arachidonate of < 10 min in 16:1-20:4 and 18:2-20:4 PtdCho and 1-3 h in 16:0-20:4, 18:1-20:4 PtdCho and PtdIns. The very short half-lives for arachidonate in the 16:1-20:4 and 18:2-20:4 PtdCho molecular species suggest important roles for these molecules in brain phospholipid metabolism and signal transduction.