IgE antibodies to omega-5 gliadin associate with immediate symptoms on oral wheat challenge in Japanese children.

BACKGROUND: Gliadins have been implicated in immunoglobulin E (IgE)-mediated allergy to ingested wheat and omega-5-gliadin is known to represent a major allergen in wheat-dependent exercise-induced anaphylaxis. Less known is whether omega-5-gliadin is a clinically relevant allergen in children with immediate allergy to ingested wheat. This study investigates whether specific IgE antibodies to omega-5-gliadin (sIgE-omega-5-gliadin-ab) could be used as a marker for oral wheat challenge outcome in wheat-sensitized children. A secondary objective was to study whether the level of sIgE-omega-5-gliadin was related to symptom severity in children with a positive challenge test. METHODS: Serum samples from 88 children sensitized to wheat, of whom 35 underwent wheat challenge, were collected consecutively. sIgE-omega-5-gliadin-ab was related to a physician's diagnosis of wheat allergy and challenge symptoms. RESULTS: The mean concentration of sIgE-omega-5-gliadin-ab was 7.25 kU(A)/l in patients with wheat allergy and 1.08 kU(A)/l in patients with no wheat allergy (P < 0.01). sIgE-omega-5-gliadin-ab was only detected in 12 of the non-wheat allergic children and 11 of them had a specific IgE to wheat below 1.30 kU(A)/l. Children reacting with severe symptoms upon challenge (n = 8) had increased levels of sIgE-omega-5-gliadin-ab compared to children with moderate, mild or no symptoms (P < 0.001). CONCLUSIONS: The presence of sIgE-omega-5-gliadin-ab is related to the reaction level to wheat challenge outcome in wheat-sensitized children. The sIgE-omega-5-gliadin-ab was found to be associated with a strong convincing history of wheat allergy also in those cases when oral food challenge was avoided. The sIgE-omega-5-gliadin-ab level may serve as a marker for clinical reactivity in wheat-sensitized individuals.

Extracts of porcine pancreas prevent progression of hypercalcemia and cachexia and prolong survival in nude mice bearing a human squamous carcinoma.

Porcine pancreatic extracts (PXs) have previously been shown to decrease blood ionized calcium in BALB/c mice (T. Yoneda, Y. Takaoka, and G. R. Mundy. FEBS Lett., 278: 171-174, 1991). In the present study, we show that the PX is effective in preventing progression of hypercalcemia and decreasing osteoclastic bone resorption associated with a human squamous carcinoma in nude mice. PX inhibited osteoclast-like cell formation in mouse bone marrow cultures and bone resorption in organ cultures of fetal rat long bones which had been stimulated by serum-free culture supernatants of this cancer. In addition, PX increased food intake, decreased weight loss, and prevented development of cachexia. In parallel with these effects, PX prolonged survival of tumor-bearing animals. PX might have therapeutic potential for management of hypercalcemia and cachexia associated with malignancy.

Molecular cloning of the cDNA encoding A + U-rich element RNA binding factor.

Using the differential display method, a new cDNA clone, termed laAUF1, encoding the human A + U-rich RNA-binding motif was isolated and sequenced. Analysis of the protein sequence of laAUF1 indicates 73% homology between the deduced polypeptide sequences of laAUF1 and AUF1 in the region encoding a consensus motif for two non-identical RNA recognition motifs (RRMs) and Gln-rich motif. We suggest that the similar affinities of laAUF1 and AUF1 for particular A + U-rich elements (ARE) sequences are related to their potencies as mRNA destabilizers.

Binding of insulin by monkey and pig hypothalamus.

Membrane preparations from monkey and pig hypothalami bound [125I]insulin specifically. The binding appeared to be greater by preparations from anterior than posterior portions of the pig hypothalamus. Binding was time dependent, and its dissociation was first order with a half-time at 22 degrees C of 14 min. Desalanine insulin was as effective as native insulin in inhibiting the binding of [125I]insulin, while proinsulin was less effective and desoctapeptide insulin still less effective in accord with their biologic activities. Binding by membranes from cortex and thalamus appeared to be less than from hypothalamus. [125I]insulin was infused into an arterial split monkey brain preparation to determine if insulin that was blood borne bound specifically to the primate hypothalamus. Half the brain was perfused with [125I]insulin alone and the other half with [125I]insulin plus an excess of unlabeled insulin. Radioautography showed specific binding of insulin localized to the median eminence, infundibular nucleus, and microvessels. Thus, the monkey and pig hypothalami bind insulin with characteristics similar to those reported for known target tissues for insulin. Furthermore, insulin from the blood stream binds to specific anatomical structures in the hypothalamus of the monkey.

Biphasic modulation by mGlu5 receptors of TRPV1-mediated intracellular calcium elevation in sensory neurons contributes to heat sensitivity.

BACKGROUND AND PURPOSE: Elevation of glutamate, an excitatory amino acid, during inflammation and injury plays a crucial role in the reception and transmission of sensory information via ionotropic and metabotropic receptors. This study aimed to investigate the mechanisms underlying the biphasic effects of metabotropic glutamate mGlu5 receptor activation on responses to noxious heat. EXPERIMENTAL APPROACH: We assessed the effects of intraplantar quisqualate, a non-selective glutamate receptor agonist, on heat and mechanical pain behaviours in mice. In addition, the effects of quisqualate on the intracellular calcium response and on membrane currents mediated by TRPV1 channels, were examined in cultured dorsal root ganglion neurons from mice. KEY RESULTS: Activation of mGlu5 receptors in hind paw transiently increased, then decreased, the response to noxious heat. In sensory neurons, activation of mGlu5 receptors potentiated TRPV1-mediated intracellular calcium elevation, while terminating activation of mGlu5 receptors depressed it. TRPV1-induced currents were potentiated by activation of mGlu5 receptors under voltage clamp conditions and these disappeared after washout. However, voltage-gated calcium currents were inhibited by the mGlu5 receptor agonist, even after washout. CONCLUSIONS AND IMPLICATIONS: These results suggest that, in sensory neurons, mGlu5 receptors biphasically modulate TRPV1-mediated intracellular calcium response via transient potentiation of TRPV1 channel-induced currents and persistent inhibition of voltage-gated calcium currents, contributing to heat hyper- and hypoalgesia.

Tissue sites of aromatization in the female rhesus monkey.

Nine female rhesus monkeys (four in the follicular and five in the luteal phases of their cycles) had catheters implanted and were infused iv with [7-3H]androstenedione (A) and [4-14C]estrone (E1) for 4 h. Blood samples were drawn at intervals from the hepatic, renal, jugular, uterine, and brachial veins and the femoral artery. The samples were analyzed for radioactivity as A and E1. The mean +/- SE MCRs for A and E1 were 280 +/- 40 and 270 +/- 30 liters/day, respectively. The mean extractions across the liver measured in six of the monkeys were 0.83 +/- 0.03 for A and 0.71 +/- 0.06 for E1. The percentage of A entering each tissue, which was measured as E1 leaving the tissue (pA,E1AV), was 0.20 +/- 0.10 for splanchnic, 0.21 +/- 0.11 for renal, 0.46 +/- 0.21 for jugular, 2.36 +/- 1.27 for arm, and 0.35 +/- 0.10 for uterine veins. Because of the sampling technique, the value for the uterus may be a reflection of ovarian blood admixture with uterine blood. There were no apparent differences in tissue aromatization between values in the follicular and luteal phases of the cycle. The overall mean value for the percentage of A infused and measured as E1 in arterial blood (pA,E1BB) was 1.01 +/- 0.38%. Using previously reported tissue blood flow, we calculate that the contributions to the overall aromatization rate of tissues drained by the brachial, renal, jugular, hepatic, and uterine veins are 23%, 5%, 5%, 4%, and 0.2%, respectively. Thus, the splanchnic tissue is a minor site for extraglandular aromatization of androgens in the rhesus monkey. An important site appears to be the arm, which reflects aromatization in adipose tissue, muscle, skin, and supporting structures.

Tissue metabolism of estrogens in the female rhesus monkey.

The overall MCRs and [rho]BB values (fraction of infused precursor measured in blood as product) and individual tissue extractions and conversions were measured in the follicular and luteal phases of normal cycling female rhesus monkeys using constant infusions of [3H]estradiol ([3H]E2) and [14C]estrone ([14C]E1). During the infusions, blood samples were obtained from the femoral artery and veins draining the splanchnic tissue, kidney, head, arm, and uterus. The overall mean (+/- SE) MCR for E2 (214 +/- 17 liters/day) was significantly less than the MCR for E1 (295 +/- 13 liters/day). There were no differences in the MCR measured in the follicular or luteal phases of the cycle. The [rho]BB values were greater for the fraction of infused E1 measured in blood as product E2 [rho]E1, E2, BB; (0.25 +/- 0.02) than for [rho] E1, E2, BB (0.11 +/- 0.01). Neither value was affected by the time of the cycle. The extractions (that fraction of steroid measured in arterial blood entering a tissue which is metabolized and not measured as that steroid in venous blood draining the tissue) across the splanchnic tissues were the largest of the tissue extractions measured (0.63 +/- 0.05 for E2 and 0.73 +/- 0.10 for estrone). The arm, uterus, kidney, and head had lower extractions, and the extraction of E2 was always lower than that of E1, probably due to the specific globulin binding of E2. Interconversion of the estrogens occurred across each tissue bed but reflected, in general, only a small portion of the total extraction, especially for the splanchnic tissue. There was no apparent difference in any of the transtissue conversions measured in the follicular as compared to the luteal phase. The administration of pharmacological amounts of dexamethasone to three monkeys resulted in a marked increase in the MCR of estradiol and a slight decrease in the MCR of estrone. Individual tissue extractions and transtissue conversions showed no consistent alteration after dexamethasone.

Porcine pancreas extract decreases blood-ionized calcium in mice and inhibits osteoclast formation and bone resorption in culture.

Patients with acute pancreatitis commonly manifest hypocalcemia for reasons which are unknown. We found that porcine pancreas extracts (PX) significantly decreased blood-ionized calcium in Balb/c mice. Partially-purified PX with a molecular mass of approximately 27 kDa decreased blood-ionized calcium in the mice. Partially-purified PX suppressed not only 45Ca release from fetal rat long bones which had been stimulated by parathyroid hormone, interleukin-1 alpha, tumor necrosis factor, transforming growth factor-alpha, 1,25-dihydroxyvitamin D3 and prostaglandin E2, but tartrate-resistant acid phosphatase-positive multinucleated cell formation in the presence of 1,25-dihydroxyvitamin D3 in mouse marrow cultures. The results suggest that there is an as yet-unidentified bone metabolism-regulating substance in porcine pancreas which might be responsible for the hypocalcemia associated with acute pancreatitis.

Interleukin-1beta induces interleukin-6 production through the production of prostaglandin E(2) in human osteoblasts, MG-63 cells.

This study was conducted to investigate the mechanism of interleukin-1beta (IL-1beta)-induced IL-6 production in human osteoblasts (MG-63 cells). Stimulation with IL-1beta resulted in the production of IL-6 and prostaglandin E(2) (PGE(2)). IL-6 production gradually increased and peaked 96 h after stimulation. IL-6 mRNA was detected between 4 and 72 h after IL-1beta stimulation. The patterns of PGE(2) production and the expression of cyclooxygenase-2 (COX-2) mRNA were biphasic after stimulation. Actinomycin D, cycloheximide, indomethacin, and NS-398 (COX-2 inhibitor) suppressed the production of IL-6 and PGE(2). Anti-PGE(2) antibody markedly reduced the production of IL-6. In addition, stimulation with 17-phenyl-PGE(2), a PGE receptor-1 (EP-1 receptor) agonist, led to the expression of IL-6 mRNA after pretreatment with IL-1beta. These findings indicate that IL-1beta-induced IL-6 production in MG-63 cells involves the following sequence of steps: IL-1beta-induced COX-2 activation, PGE(2) production, and EP-1 receptor signaling prior to IL-6 production.

The effect of a TXA2 receptor antagonist ON-579 on experimental allergic reactions.

The effect of a thromboxane A2 (TXA2) receptor antagonist, ON-579, on experimental allergic skin and airway reactions was studied in vivo. ON-579 at doses of 1 and 20 mg/kg clearly inhibited U-46619-induced increases in respiratory resistance (Rrs) in guinea pigs. ON-579 at doses of 1, 20 and 50 mg/kg inhibited the aerosolized antigen-induced biphasic increase in Rrs in guinea pigs. Moreover, ON-579 clearly inhibited repeated aeroantigen-induced airway hyperreactivity in guinea pigs. ON-579, however, did not have any significant effects on allergic cutaneous reactions in rats. These results suggest that ON-579 is a relatively selective TXA2 antagonist, especially in the airways, and indicate the efficacy of ON-579 on antigen-induced increase in airway resistance and antigen-induced airway hyperreactivity in guinea pigs.

The metabolic clearance rate, head and brain extractions, and brain distribution and metabolism of progesterone in the anesthetized, female monkey (Macaca mulatta).

The brain distribution and metabolism of progesterone were studied in female, rhesus monkeys. Adult monkeys were anesthetized with ketamine and were given a constant infusion of [3H]- or [14C] progesterone. Blood samples were obtained from cannulae inserted into the carotid artery, the jugular vein and lateral (transverse) sinus. The metabolic clearance rate of progesterone was 295 +/- 49 (S.E.) 1/day. The head extraction of progesterone was 30.4 +/- 8.3% (S.E.) and the brain extraction 26.0 +/- 9.18% (S.E.). The peripheral conversion ratios of progesterone to 20alpha-hydroxypregn-4-en-3-one (20alpha-OHP) and 5alpha-pregnan-3,20-dione (5alpha-DHP) were 10.0 +/- 1.3% (S.E.) and 2.4 +/- 0.3% (S.E.), respectively. These same conversion ratios for across the head were 4.8 +/- 1.0% (S.E.) and 1.5 +/- 0.6% (S.E.) and for across the brain 5.0 +/- 0.7% (S.E.) and 2.2 +/- 0.6% (S.E.). The concentration of radioactive progesterone was 2-5 times higher in brain tissues compared to the carotid arterial blood. The tissue concentrations of radioactive progesterone compared to a cerebrum 'control' sample: was lower in the central gray (P less than 0.05); were the same for the amygdala, hippocampus, preoptic-anterior hypothalamus, cerebellum, hypothalamus, thalamus, and anterior pituitary; and were higher in the cervical spinal cord, optic chiasm, mesencephalon, medulla oblongata and pons (P less than 0.1). The distribution pattern of 20alpha-OHP formed from progesterone was similar to that of progesterone. 5alpha-DHP formed from progesterone had a different distribution than progesterone, being highest in the central gray area. High concentrations of 5alpha-DHP were also observed in the mesencephalon, medulla oblongata and hypothalamus and low values in the anterior pituitary. Infusions of [3H]20alpha-OHP and [3H]5alpha-DHP were used to evaluate the in vivo metabolism of progesterone by different brain areas. [3H]Estradiol infused into one monkey had its highest concentration in the anterior pituitary which was 20 times higher than in the carotid arterial blood.

The use of zona-free aged unfertilized human oocytes as a predictor for successful subzonal insemination.

OBJECTIVE: To examine the reliability of the zona-free aged (1 day old) human unfertilized oocyte sperm penetration assay for assessing sperm fertilizing ability and to determine the predictive value of this assay for subsequent subzonal insemination (SUZI) outcomes. DESIGN: A total of 253 unfertilized oocytes from total fertilization failure patients and from good fertilization rate (> 70%) patients in standard IVF were inseminated with donors' spermatozoa, and penetration rates were compared in each group. Two hundred seventy-two unfertilized oocytes from total fertilization failure, poor fertilization (< 30%), and normal fertilization (> 30%) were inseminated with husbands' spermatozoa, and penetration rates were compared between the three groups. In 29 patients, the results of the zona-free assay performed in previous IVF were compared with the fertilization rates of subsequent SUZI. RESULTS: In the zona-free assay using donors' spermatozoa, there was no difference in penetration rates between the two groups (109/122, 89.3% versus 114/131, 87.0%). Penetration rates using partners' spermatozoa were positively correlated with fertilization rates in standard IVF (total fertilization failure 34/75, 45.3%; poor fertilization 56/77, 72.7%; normal fertilization 108/120, 90.0%). There was a significant difference in fertilization rates after SUZI between the patients with negative and positive penetration in zona-free assay (4/53, 7.5%, versus 54/174, 31.0%). CONCLUSION: The zona-free human oocyte assay may primarily reflect sperm fertilizing ability. This asssay also could be a reliable predictor for subsequent SUZI outcome.

Effects of low concentrations of nitric oxide on the zona pellucida binding ability of human spermatozoa.

OBJECTIVE: To investigate the direct effects of sodium nitroprusside, a nitric oxide donor, on the fertilizing ability of human spermatozoa in vitro. DESIGN: Prospective, controlled in vitro study. SETTING: IVF Unit, Medical College Hospital. PATIENT(S): Women undergoing conventional IVF. INTERVENTION(S): Human spermatozoa samples were incubated with a nitric oxide donor and a nitric oxide quencher, carboxy-imidazolineoxyl N-oxides. MAIN OUTCOME MEASURE(S): The capacitation and the acrosome reaction rates were determined by chlortetracycline assay. Sperm zona pellucida binding and sperm penetration into oocytes were determined using the hemizona assay and the human aged zona-free oocyte sperm penetration assay. RESULT(S): High concentrations of sodium nitroprusside (10(-3) and 10(-5) M) inhibited sperm motility and viability. In contrast, low concentrations of sodium nitroprusside (10(-7) and 10(-8) M) did not affect motility and resulted in increased capacitation (47 +/- 6% at 10(-7) M, 42 +/- 6% at 10(-8) M, and 24 +/- 4% in controls, respectively, P < 0.01). A twofold increase in the hemizona index occurred compared to the matched control. However, low levels of sodium nitroprusside treatment did not affect the acrosome reaction and human zona-free oocyte sperm penetration rates. CONCLUSION(S): Low concentrations of nitric oxide may have some physiologic role in fertilization through the enhancement of capacitation and zona pellucida binding but not by the induction of the acrosome reaction or the facilitation of penetration into oocytes.

Autoradiographic localization of estradiol- and progesterone-concentrating neurons in the isolated rhesus monkey hypothalamus.

Progesterone- and estradiol-concentrating neurons were autoradiographically localized in the in situ, vascularly isolated, rhesus monkey hypothalamus. Estradiol-concentrating neurons were dispersed throughout the hypothalamus, their density being greatest in the medial preoptic (MPOA) and medial basal hypothalamic nuclei (viz. dorsomedial, ventromedial (VMH) and infundibular nuclei). In contrast, progesterone-concentrating neurons were less densely localized in the medial preoptic, ventromedial (medial division) and infundibular nuclei. There was a virtual absence of progestin-concentrating cells in the anterior and posterior hypothalamic zones. This localization of cells was not attributed to the perfusion pattern of the isolated hypothalamus since a complete distribution of injected microspheres was found throughout the hypothalamus. The results of these studies indicate that a possible division between estradiol- and progesterone-concentrating neurons exists within the MPOA-medial basal hypothalamic nuclei of the rhesus monkey.

A "PICA communicating artery" aneurysm: case report.

We present an unusual case of an aneurysm of the distal posterior inferior cerebellar artery (PICA). The aneurysm was associated with a unilateral PICA that supplied both cerebellar hemispheres and arose from an anastomotic vessel to the contralateral circulation, a branch of the contralateral PICA. Such an aneurysm has not been reported previously. The associated of vascular anomalies with aneurysms of the PICA is discussed.

The model of arthritis induced by superantigen in mice.

Subcutaneous injection of Staphylococcal enterotoxin B (SEB) produced by Staphylococcus aureus, caused severe arthritis in DBA/1J mice which had been previously immunized with bovine type II collagen. The severity of this arthritis was dose dependent and prolonged joint inflammation with erosion of bone was observed. Anti- type II collage antibodies were detected in the serum of arthritic mice. Effector T cells against type II collagen were also detected by means of delayed type hypersensitivity in the skin. Moreover, a significant decrease in the ratio between T cells and B cells and an increase in the ratio between CD4+ cells and CD8+ cells was observed in spleen cells from arthritic mice. Prednisolone suppresses the induction and development of clinical signs of arthritis in mice. This evidence suggests that this experimental arthritis model may provide a means to examine the role of superantigens and the efficacy of pharmacological agents for the treatment of rheumatoid arthritis.

A high performance isolated rat brain preparation. Part I: Operative technique and recipient extracorporeal perfusion.

The microsurgical preparation and high level performance with extracorporeal recipient perfusion of a stable, totally isolated rat brain model is presented. Emphasis has been placed on the use of the operating microscope, bipolar cautery and mechanical fixation units to affect a complete ablation of all non-osseous cephalic and vertebral tissues with minimal physiological disturbance of the animal as a whole. The simplified technique of recipient support circulation of the isolated brain organ achieves at least 5 h of tissue viability and function as measured by electrocortical activity, A-Vo2 differences and morphological appearance. To date, no other isolated rodent brain model has been described which is truly anatomically separated from all cephalic tissues. Additionally, other surgical attempts at isolation have required the use of abnormal physiological states, including deep hypothermia and circulatory arrest.

A high performance isolated rat brain preparation. Part II. Cerebral blood flow in relation to perfusion pressure and cerebral hypothermia.

The effects of changes of perfusion pressure and hypothermia on the cerebral blood flow of a new isolated rat brain preparation have been studied in 7 animals. The cerebral blood flow was extremely sensitive to the mean arterial pressure of perfusion, showing little evidence of autoregulation at low pressure, and only slight evidence in hypertension. The cerebral blood flow was lower in hypothermia. The preparation maintained good electroencephalographic activity throughout, and arteriovenous differences persisted up to 1 h after it became isoelectric.

The role of EC-IC in the treatment of giant intracranial aneurysms.

Extracranial to intracranial bypass surgery was used in 27 cases of intracranial giant aneurysm to prevent ischemic complications. In 19 of 21 patients the aneurysm was considered unfit for a direct clipping or ligation, and an EC-IC bypass was done in conjunction with staged clipping of the internal carotid artery or occlusion of the middle cerebral artery. All bypass grafts have remained patent postoperatively. None of the patients have developed ischemic complications in the 6--14 month period of follow-up.