Lettuce anaphylaxis: identification of a lipid transfer protein as the major allergen

BACKGROUND: Allergy to plant-derived foods is associated with birch pollinosis in central and northern Europe. Symptoms elicited are usually limited to the oropharyngeal system. By contrast, in the Mediterranean area, allergy to the same foods manifests more frequently with systemic reactions caused by nonspecific lipid transfer proteins (nsLTP), independently of an associated pollinosis. OBJECTIVE: We sought to investigate the pattern of immunoglobulin E (IgE) binding protein bands implicated in lettuce allergy, in particular the presence of an nsLTP. METHODS: Consecutive lettuce allergic patients were selected. Determination of serum-specific IgE, immunoblot, and inhibition experiments were performed in order to study the pattern of IgE binding proteins and the potential cross-reactivity to pollens. Inhibition studies with recombinant allergens were conducted to identify the lettuce allergens. The major allergen was subjected to N-terminal amino acid sequencing. RESULTS: Fourteen patients were diagnosed as being allergic to lettuce. All were sensitized to Platanus pollen. Ten of them showed specific IgE to a lettuce protein of 9-kDa. The IgE binding to this protein was completely inhibited by the cherry-LTP and peach extract. The N-terminal sequence of the 9-kDa protein showed a high degree of amino acid sequence identity to other nsLTPs. A clear partial cross-reactivity was observed between lettuce-LTP and Platanus-pollen extract. CONCLUSIONS: An LTP has been demonstrated to be a major allergen in patients suffering from lettuce allergy.     

Excitability of reciprocal and recurrent inhibitory pathways after voluntary muscle relaxation in man

Summary We studied the potential contribution of postsynaptic mechanisms to the depression of reflex excitability which occurs immediately after a voluntary release from tonic muscle contraction. The excitability of the Soleus (Sol) motor pool was tested at rest and after voluntary muscle relaxation. In both cases the Sol H-reflex was conditioned by 1. a single shock to the peroneal nerve, in order to activate the Ia interneurones (INs) mediating the reciprocal inhibition via a peripheral input, or by 2. a short-lasting voluntary contraction of the Tibialis Anterior (TA) muscle, to activate the Ia INs via a central command. Changes in excitability of Renshaw cells were also tested at rest and after release, to assess the role of recurrent inhibition in the release-induced inhibition of the Sol H-reflex. It was demonstrated that: 1. the excitability of the INs mediating the reciprocal inhibition was only slightly enhanced in comparison with resting conditions; 2. the H-reflex of the antagonist muscle (TA) evoked after Sol release was not consistently facilitated with respect to rest; 3. the command to contract the TA muscle reduced the H-reflex of the Sol muscle during rest but not after Sol release; 4. recurrent inhibition did not increase its effect in the post-release period. Such features suggest that recurrent and reciprocal post-synaptic inhibitions do not play a major role in reducing the reflex excitability of a relaxing muscle; rather, the command to release prevents the reciprocal inhibitory effect which accompanies the contraction of the antagonist muscle. The findings support the concept that release-induced reflex depression is mediated mainly by presynaptic inhibition of autogenetic spindle afferences (Schieppati and Crenna 1984).Supported by Italian M.P.I.     

Immunochemical characterization of mouse brain-associated theta (Thy-1) antigen.

The Thy-1 antigens or rat brain and thymus have been isolated and chemically characterized, but those of mice have not been identified. Moreover, it is uncertain whether the antigens are glycolipids or glycoproteins. This study with highly purified preparations of gangliosides GM₁, ¹GD_1a, GD_1b and GT_1b from bovine brain and several ganglioside fractions from mouse brain showed that Thy-1 activity does not reside in gangliosides, but rather in the chloroform-methanol-insoluble residue of brain remaining after extraction of gangliosides. The antigen could be solubilized from this residue with a non-ionic detergent. The antigenic activity of the solubilized preparation was heat-labile but resistant to periodate. The chemical properties of the Thy-1 antigen of mouse brain are discussed.     

Activity of human erythrocyte gangliosides as a receptor to HVJ

Liposomes could bind and fuse efficiently to human erythrocytes in the presence of HVJ when they contained gangliosides isolated from human erythrocytes. Sialosylparagloboside, which has a terminal sequence of NeuAcα2?3Ga1β1?4GlcNac, has a much higher receptor activity to the virus than GD_1a, GD_1b, GT_1b, and GT_1a, all of which contain the terminal sequence of NeuAcα2?3Galβ1?3GalNAc or NeuAcα2?8NeuAcα2?3Galβ1?3GalNAc. The activity of sialosylparagloboside is comparable to that of glycophorin, a major sialoglycoprotein of human erythrocytes, when compared on the basis of the required amount (as sialic acid) of compounds. The high affinity of sialosylparagloboside to the viral HANA protein is also suggested by the finding that it showed high inhibitory activity against HVJ-mediated binding of glycophorin liposomes to erythrocytes. Sialosylparagloboside was also highly susceptible to the viral sialidase, the other biological function of HANA protein.     

Effects of manipulating catecholamines on the incidence of the preovulatory surge of of luteinizing hormone and ovulation in the rat: evidence for a necessary involvement of hypothalamic adrenaline in the normal or 'midnight' surge

The preovulatory surge of luteinizing hormone reaches a maximum at 18.00 h on the day of pro-oestrus in female rats maintained with regular lighting from 06.00 to 20.00 h. This surge is initiated by a discharge of luteinizing hormone-releasing hormone into hypophysial portal blood. In this study, drugs which affect catecholamine-mediated neurotransmission were administered on the day of pro-oestrus and the effects on serum concentrations of luteinizing hormone and on subsequent ovulation were observed. alpha-Methyl-p-tyrosine, diethyldithiocarbamate and SKF 64139 inhibit catecholamine synthesis at the level of tyrosine hydroxylase, dopamine beta-hydroxylase and phenylethanolamine N-methyltransferase, respectively. Although alpha-methyl-p-tyrosine suppressed ovulation, it had a negligible effect on the incidence of the preovulatory surge. In contrast, the various treatments with diethyldithiocarbamate and SKF 64139 resulted in a minimal occurrence of the 18.00 h surge; at relatively low doses, however, these drugs frequently elicited a surge at 22.00 or 24.00 h which invariably resulted in ovulation. The failure of the surge after diethyldithiocarbamate or SKF 64139 was not associated with a loss of pituitary sensitivity to luteinizing hormone-releasing hormone. In terms of the hypothalamic concentration of dopamine, noradrenaline, adrenaline and 5-hydroxytryptamine at 18.00 h on pro-oestrus, the only common effect of diethyldithiocarbamate and SKF 64139, given in a dose which blocks the surge, was a severe depletion of adrenaline; alpha-methyl-p-tyrosine failed to produce this effect despite inducing a marked depression of dopamine and a moderate loss of noradrenaline. Neither the increase in hypothalamic dopamine after diethyldithiocarbamate, nor the alpha 2 receptor blocking properties of SKF 64139 appear to be relevant in this context since injections of L-dopa or piperoxane, an alpha 2 receptor antagonist, were without effect on the surge or ovulation. The failure of the surge after prazosin, an alpha 1 receptor antagonist, indicates that the function of adrenaline may be mediated postsynaptically by alpha 1 receptors. Clonidine, an alpha 2 receptor agonist which reduces the turnover rate of hypothalamic adrenaline, had effects of the surge and ovulation which were comparable to those of diethyldithiocarbamate and SKF 64139, the relatively low doses causing some of the surges to occur at 24.00 instead of 18.00 h and higher doses suppressing the surge at both times and thus preventing ovulation.(ABSTRACT TRUNCATED AT 400 WORDS)     

Inhibition of adhesion of enterotoxigenic Escherichia coli cells expressing F17 fimbriae to small intestinal mucus and brush-border membranes of young calves

Enterotoxigenic Escherichia coli strains expressing F17 fimbriae bind to the intestinal mucosa of young calves. F17 fimbriae recognize receptors present in the mucus layer and the brush-border membranes from duodenum, jejunum and ileum. The adhesion of E. coli F17 can be inhibited by several glycoproteins. Adhesion is also inhibited by pretreatment of mucus and brush-border membranes with sodium metaperiodate. The use of glycoconjugates as potential adhesion-blockers is further discussed.     

Inhibition of proliferative responses of mouse spleen lymphocytes by bovine milk κ‐casein digests

The inhibitory effects of bovine milk κ‐casein and its enzymatic digests on the proliferative responses of mouse spleen lymphocytes induced or not induced by mitogens were studied with a colorimetric assay using 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide (MTT). κ‐Casein and its glyco‐macropeptide (residues 106–169) inhibited the lipopolysaccharide (LPS)‐induced proliferative response, but the inhibitory effect was lost significantly after neuraminidase and chymotrypsin digestions. In contrast, trypsin and pronase digestions of K‐casein increased inhibitory effects. The pronase digest inhibited the proliferative responses not only induced by LPS but also in the absence of mitogen or when induced by concanavalin A and phytohemagglutinin. The pronase digest seemed to possess weak cytotoxity for lymphocytes. The inhibitory peptide was a glycopeptide(s) having specific size, and the inhibitory effects were reduced significantly by neuraminidase and chymotrypsin digestions. Moreover, similar inhibitory effects on proliferation of lymphocytes were observed in pronase‐digested bovine milk αs₁‐casein and β‐casein. These findings suggest that some peptides produced from milk caseins by the action of gastrointestinal proteinases might contribute to down‐regulate the immune response of neonates.     

Supplementation Effect of Early Pregnancy Factor-Positive Serum into Bovine In Vitro Fertilization Culture Medium

PROBLEM: Early pregnancy factor (EPF) has been detected in pregnant bovine serum, and its activity appeared from 24 to 48 hr after insemination. However, in bovine in vitro fertilization (IVF), an EPF-like substance(s) had been detected in the culture medium of fertilized ovum. Therefore, we think that EPF and EPF-like substance(s) are very important materials for the development of the embryo. In this study, we examined the development of the embryo when fertilized bovine ova were cultured with IVF culture medium supplemented with EPF-positive or -negative serum. METHOD OF STUDY: EPF activity of each serum (fetal calf serum [FCS], calf serum [CS], estrus bovine serum, and pregnant bovine serum) was assessed by the bovine-rosette inhibition test. The sera were supplemented in TCM-199 culture medium, and IVF bovine ova were cultured with the media for 6 or 7 days, respectively. The culture media of each group were evaluated for EPF activity by the bovine-rosette inhibition test 48 hr after IVF. The cleavage rate of each group was calculated at 48 hr, and 6 or 7 days after IVF. The culture medium of cumulus cells was also tested for EPF activity. RESULTS: Only the pregnant bovine sera were EPF positive. All the culture media 48 hr after IVF became EPF positive. Additionally, the culture medium of cumulus cells did not have EPF activity. There was no significant difference in the cleavage rate of the EPF-positive and - negative sera 48 hr after IVF. However, the cleavage rate of EPF-positive sera tended to be higher than the negative sera. In contrast, the blastocyst development rates of EPF-positive sera were significantly higher than those of the negative sera 6 to 7 days after IVF (P < 0.05). CONCLUSIONS: The data suggest that an EPF-like substance(s) may be secreted from the in vitro fertilized bovine ovum but not from the cumulus cell, and that the EPF in the pregnant serum may accelerate the development of the bovine embryo in IVF.     

Neuronal histamine inhibits methamphetamine-induced locomotor hyperactivity in mice

Mianserin (5-20 mg/kg), like chlordiazepoxide (2.5-10 mg/kg), inhibits the shock-induced suppression of drinking (SSD) in rats. However, in contrast to chlordiazepoxide, the effect of mianserin is not blocked by the benzodiazepine antagonist, Ro 15-1788 (10 mg/kg). Although mianserin does not inhibit [3H]diazepam binding in vitro it has now been found to enhance [3H]flunitrazepam binding to mouse whole brain in vivo at 10-100 mg/kg p.o. These results suggest that mianserin does influence central benzodiazepine receptors, but the mechanism by which it does so differs from that of chlordiazepoxide.     

Transcerebellar inhibitory interaction between the bilateral vestibular nuclei and its modulation by cerebellocortical activity

Summary In decerebrate, unanesthetized cats, the brain stem was longitudinally cut at the midline from its dorsal to ventral surface with the cerebellum kept intact, eliminating neural interactions between the bilateral vestibular nuclei through the brain stem.Extracellular spike potentials of vestibular type I neurons identified by horizontal rotation were distinctly inhibited by contralateral vestibular nerve stimulation. This crossed inhibition was abolished by removal of the medial part of the cerebellum, indicating that the inhibition was mediated through the cerebellum. Neither aspiration of the flocculus on the recording side nor intravenous administration of picrotoxin eliminated transcerebellar crossed inhibition, suggesting that it is mediated through the cerebellar nuclei. When the fastigial, interposite and dentate nuclei were stimulated, inhibition of vestibular type I neurons was produced only from the contralateral fastigial nucleus. Cerebellocortical stimulation which inhibited fastigial type I neurons suppressed transcerebellar crossed inhibition. Effective sites for suppression of transcerebellar crossed inhibition were localized to lobules VI and VIIa in the vermal cortex on the side of labyrinthine stimulation.Intracellular recordings were made from type I neurons in the medial vestibular nucleus. Stimulation of the contralateral vestibular nerve and the contralateral fastigial nucleus produced IPSPs in these neurons with the shortest latency of 3.8 msec and 1.8 msec, respectively. The difference between these two latency values approximates the shortest latency of spike initiation of fastigial type I neurons in response to vestibular nerve stimulation. It is postulated that transcerebellar crossed inhibition is mediated through the fastigial nucleus on the side of labyrinthine stimulation.