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NPT has a high sensitivity and specificity in the diagnosis of AIA. An oral provocation should be performed to confirm the result whenever the clinical situation of the patient permits it.

A study was carried out in the Castilla y León region of Spain to investigate the presence of Escherichia coli O157:H7 in raw ewe's milk samples collected from several cheese factories during 1 year. All specimens were examined for E. coli O157:H7 by selective enrichment at 41.5 +/- 1.0 degrees C, after both 6 and 22 h of incubation, and then immunomagnetically separated and plated on cefixime-potassium tellurite-sorbitol MacConkey agar. No growth was obtained in the enrichment broth after a 6-h incubation. Presumptive colonies obtained after 22 h of incubation were screened by a multiplex PCR assay for the presence of rfbO157 and fliCH7 genes. Of all the ewe's milk samples studied, three were positive for E. coli O157:H7. The E. coli O157:H7 strains that were positive for the rfbO157 and fliCH7 genes were then analyzed by multiplex PCR for the presence of virulence genes (stx1, stx2, ehxA, and eaeA). All E. coli O157:H7 isolates were Shiga toxigenic and harbored additional genes related to virulence (ehxA and eaeA). The predominant Stx toxin type was stx2. These results demonstrate that raw ewe's milk used in cheesemaking may be sporadically contaminated with E. coli O157:H7 strains that are potentially pathogenic for humans.

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A partial cDNA encoding most of the third intracellular loop of the mouse a, d -adrenergic receptor subtype was amplified from hippocampus by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligodeoxynucleotide primers . This DNA fragment was used as a probe to isolate an a, d -adrenergic receptor cDNA from a mouse brain cDNA library . The deduced amino acid sequence encodes a potential protein of 562 amino acids, and northern hybridization of poly(A)+ RNA isolated from mouse brain detected a single 3 .0-kb transcript . Partial cDNA fragments of the alband a, a -adrenergic receptor subtypes were also amplified from mouse brain and sequenced . Analysis of the mRNA expression by RT-PCR indicated that the a,-adrenergic receptors are widely distributed in mouse tissues . The a,d subtype is expressed in brain areas such as hippocampus, striatum, and brainstem and also in many extracerebral tissues, such as lung, liver, heart, kidney, and spleen . The a,a subtype is also expressed in many tissues, whereas the alb subtype has a more restricted expression, with high levels in striatum, brainstem, and diencephalus . Key Words : a,d -Adrenergic receptor-Mouse-cDNA cloning-Tissue distribution -a,-Adrenergic receptor subtypes . J. Neurochem . 65, 2387Neurochem . 65, -2392Neurochem . 65, (1995 .The adrenergic receptors are members of a large family of membrane proteins that are coupled to guanine nucleotide-binding proteins (G proteins) . They are currently divided into three types (a,, a2 , and ß), each having several receptor members (Lomasney et al., 1991a ;Bylund, 1992) . Radioligand binding data with the a,-adrenergic receptor agonists oxymetazoline and methoxamine and the antagonists WB4101 and phentolamine indicated the existence of two a,-adrenergic receptor subtypes in rat cerebral cortex, which were named a1A and a,B (Battaglia et al., 1983 ; Morrow and Creese, 1986;Han et al., 1987) .Several cDNAs and genes encoding a t-adrenergic receptor subtypes from various sources, and initially named a]A/alD, a,B , and a lc , have been isolated by