Sensitization to acid-hydrolyzed wheat protein by transdermal administration to BALB/c mice, and comparison with gluten.

BACKGROUND: An increasing number of studies have shown that hydrolyzed wheat protein (HWP) can induce IgE-mediated hypersensitivity by skin contact and/or food ingestion. However, there has been no study of the sensitizing potential of HWP. In this study, the possibility of transdermal pathway for sensitization to acid-HWP (HWP1) was investigated using BALB/c mice, and compared with that of gluten. METHODS: HWP1 or gluten (500 µg/mouse) was transdermally administered using patches. After three or four cycles of sensitization for 3 days/week, active systemic anaphylaxis (ASA) was induced by intraperitoneal injection of the antigen, and rectal temperatures, scores of anaphylactic responses, and plasma histamine levels were determined. Because HWP1 was included in facial soap in Japan, the effect of detergent on the sensitizing potential was also investigated. RESULTS: Transdermal administration of HWP1 induced dose-dependent production of IgE and IgG1. After sensitization for 3 or 4 weeks, intraperitoneal injection of HWP1 caused ASA, leading to decreased rectal temperatures, increased anaphylaxis scores, and increased plasma histamine levels. In addition, splenocytes harvested after ASA produced IL-4, IL-5, and IL-10 by re-stimulation with HWP1. Transdermal exposure to gluten also induced IgE and IgG1 production, and intraperitoneal injection of gluten also induced ASA only in mice sensitized in the presence of sodium dodecyl sulfate. CONCLUSIONS: Transdermal exposure to HWP1 is sufficient to activate key immune pathways necessary for sensitizing mice for immediate hypersensitivity reactions. This study shows that HWP has a sensitizing potential as well as gluten, whereas its allergenicity may be different from that of gluten.

Synaptotagmin II negatively regulates Ca2+-triggered exocytosis of lysosomes in mast cells.

Synaptotagmins (Syts) I and II are believed to act as Ca2+ sensors in the control of neurotransmission. Here we demonstrate that mast cells express Syt II in their lysosomal fraction. We further show that activation of mast cells by either aggregation of FcepsilonRI or by Ca2+ ionophores results in exocytosis of lysosomes, in addition to the well documented exocytosis of their secretory granules. Syt II directly regulates lysosomal exocytosis, whereby overexpression of Syt II inhibited Ca2+-triggered release of the lysosomal processed form of cathepsin D, whereas suppression of Syt II expression markedly potentiated this release. These findings provide evidence for a novel function of Syt II in negatively regulating Ca2+-triggered exocytosis of lysosomes, and suggest that Syt II-regulated secretion from lysosomes may play an important role in mast cell biology.

Inhaled corticosteroids stabilize constrictive bronchiolitis after hematopoietic stem cell transplantation.

Post transplantation constrictive bronchiolitis (PTCB) is the most common pulmonary complication among long-term survivors of allogeneic hematopoietic stem cell transplantation (HSCT). It is a late manifestation of GVHD. Its treatment with high-dose systemic corticosteroids and other immunosuppressive regimens is associated with multiple side effects. Topical corticosteroids are used for the treatment of other manifestations of GVHD to minimize these side effects. We conducted a retrospective analysis of a series of adult patients to evaluate the efficacy of high-dose inhaled corticosteroids in the treatment of PTCB. Seventeen patients with new-onset airflow obstruction were diagnosed with PTCB. Their forced expiratory volume in 1 s (FEV1) declined from a median of 84% (range, 56-119) before HSCT to 53% (26-82) after HSCT. All patients received inhaled fluticasone propionate 500-940 microg two times daily. Symptoms of airway obstruction improved and FEV1 stabilized 3-6 months after treatment. We conclude that high-dose inhaled corticosteroids may be effective in the treatment of PTCB and propose a plausible mechanism of its action. A prospective evaluation of its efficacy is warranted.

Description of International Caenorhabditis elegans Experiment first flight (ICE-FIRST).

Traveling, living and working in space is now a reality. The number of people and length of time in space is increasing. With new horizons for exploration it becomes more important to fully understand and provide countermeasures to the effects of the space environment on the human body. In addition, space provides a unique laboratory to study how life and physiologic functions adapt from the cellular level to that of the entire organism. Caenorhabditis elegans is a genetic model organism used to study physiology on Earth. Here we provide a description of the rationale, design, methods, and space culture validation of the ICE-FIRST payload, which engaged C. elegans researchers from four nations. Here we also show C. elegans growth and development proceeds essentially normally in a chemically defined liquid medium on board the International Space Station (10.9 day round trip). By setting flight constraints first and bringing together established C. elegans researchers second, we were able to use minimal stowage space to successfully return a total of 53 independent samples, each containing more than a hundred individual animals, to investigators within one year of experiment concept. We believe that in the future, bringing together individuals with knowledge of flight experiment operations, flight hardware, space biology, and genetic model organisms should yield similarly successful payloads.

Atrial natriuretic peptide modulates alveolar type 2 cell adenylyl and guanylyl cyclases and inhibits surfactant secretion.

Alveolar epithelial type 2 (T2) cells isolated from the lungs of adult rats responded to exogenous atrial natriuretic peptide (ANP) by two signalling mechanisms. First, ANP induced a dose-dependent reduction of ligand-stimulated adenylyl cyclase activity and cAMP accumulation. This effect was inhibited by the addition of GDPbetaS or by pretreatment with pertussis toxin (PT), consistent with mediation by a Gi protein(s). PT-catalyzed [32P]ADP-ribosylation, immunoblots with specific antisera, and Northern blot analysis demonstrated that T2 cells contain the G-proteins Gi2 and Gi3 which could transduce this signal. ANP also promoted PT-insensitive, dose-dependent accumulation of cGMP, consistent with activation of a receptor guanylyl cyclase. Isoproterenol-stimulated phosphatidylcholine secretion was markedly attenuated by ANP, and this effect was inhibited by PT pretreatment, consistent with mediation by a Gi protein(s). These data indicate that in addition to the lung being a major clearance organ for circulating ANP, lung parenchymal cells are targets of ANP action.

U73122 inhibits the dephosphorylation and translocation of cofilin in activated macrophage-like U937 cells.

Cofilin, an actin-binding protein, plays an important role in the migration, phagocytosis, and superoxide production of activated phagocytes through cytoskeletal reorganization. In unstimulated phagocytes, cofilin is a major phosphoprotein. However, upon activation, the phosphoprotein is dephosphorylated and translocated from cytosol to plasma membranes. Only the unphosphorylated form of cofilin is an active form that binds actin, whereas the regulatory mechanisms of cofilin have not been elucidated. We found that 1-[6-[[17beta-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione (U73122), an inhibitor of phospholipase C (PLC), suppressed both opsonized zymosan (OZ)-induced dephosphorylation and translocation of cofilin in macrophage-like U937 cells at 4 microM concentration. OZ triggered an increase in inositol 1,4,5-trisphosphate (IP3), and U73122 inhibited it. 1-[6-[[17beta-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-pyrrodione-dione (U73343), which was employed as an inactive analogue, had no such inhibitory activities as did U73122. Furthermore, herbimycin A, an inhibitor of src-type tyrosine kinase, also inhibited OZ-triggered IP3 formation. These results suggest that the activity and localization of cofilin are regulated by PLC at the downstream of src-family tyrosine kinase.

Effects of adenosine and 2'-deoxyadenosine on epidermal keratinocyte proliferation: its relation to cyclic AMP formation.

Although it has been reported that adenosine has an inhibitory effect on keratinocyte proliferation at both G2 and S phases of the cell cycle, its relation to cyclic AMP formation through the adenylate cyclase system has been less well characterized. In order to determine the precise mechanism of the adenosine effect, another physiologic adenine nucleoside, 2'-deoxyadenosine was employed. 2'-Deoxyadenosine was shown to be remarkably different from adenosine in its ability to stimulate the epidermal adenylate cyclase; whereas adenosine markedly increased cyclic AMP levels of pig epidermis, deoxyadenosine had a much weaker effect on the cyclic AMP levels of the skin. Using several parameters of cell proliferation, comparison was made between the effects of these two compounds. Pig keratinocyte explant culture system was employed for the measurement of outgrowth and mitosis. Mitosis was determined after 72-h incubation (to monitor the overall cell proliferation inhibition) and 4-h incubation (to monitor G2 phase inhibition) with the chemicals. Pig skin keratome slice system was employed for [3H]thymidine uptake measurement. Both adenosine and deoxyadenosine were shown to have marked inhibitory effects on keratinocyte out-growth, [3H]thymidine uptake, and keratinocyte mitosis. The effects of deoxyadenosine on outgrowth and [3H]thymidine uptake were greater than that of adenosine. The inhibitory effect of adenosine and deoxyadenosine on mitosis were about the same in both 4-h and 72-h incubation systems. Thus deoxyadenosine, which is a much weaker stimulator of epidermal adenylate cyclase, was also shown to be as potent an inhibitor of keratinocyte proliferation as adenosine. These results further substantiate the view that cyclic AMP elevating agents (such as adenosine and deoxyadenosine) might not necessarily reveal their inhibitory effects on keratinocyte proliferation through their effects of cyclic AMP formation.

The effects of thyrotropin-releasing hormone on the endocrine pancreas.

TRH has been shown to be present in the pancreas. To examine a possible role for TRH in the control of endocrine pancreatic function, we have studied the effects of TRH on the isolated perfused rat pancreas preparation. Arginine caused release of TRH from the preparation. The mean maximum TRH peak was 85 +/- 12 pg/ml and occurred later than the first phase of glucagon release. Glucagon (2000 pg/ml) did not release TRH from the preparation. There was no detectable basal release of TRH. Glucose did not stimulate release of TRH from the pancreas preparation. TRH (10 ng/ml) by itself had no effect on insulin or glucagon release. TRH enhanced arginine-induced glucagon release; mean summated glucagon was 8228 +/- 1138 (SE) pg/ml compared to controls (4530 +/- 447 pg/ml; P less than 0.01). There was a tendency for TRH to enhance second phase glucose-induced insulin release. Pancreatic physiology is in part regulated by locally acting hormones and TRH may be one of these hormones.

Participation of cofilin in opsonized zymosan-triggered activation of neutrophil-like HL-60 cells through rapid dephosphorylation and translocation to plasma membranes.

We studied the roles of cofilin, an actin-binding phosphoprotein, in superoxide production of neutrophil-like HL-60 cells triggered by opsonized zymosan (OZ). OZ caused dephosphorylation of cofilin as well as a transient increase of F-actin. Both reactions were complete within 30 s. Okadaic acid (OA) magnified the OZ-triggered O2--production 3.3-fold at 1 microM, but inhibited it completely at 5 microM. We used these critical concentrations to study the effects of OA on changes in phosphorylation and intracellular localization of cofilin. The OZ-induced dephosphorylation of cofilin was inhibited by 5 microM OA but not by 1 microM OA. Subcellular fractionation and immunoblotting revealed that 1 microM OA increased cofilin on the phagosomal membranous fraction but 5 microM OA decreased it. At 1 microM, OA increased translocation of p47phox to membranes, which may explain in part the enhancing effect of 1 microM OA. Confocal laser scanning microscopy showed that: (i) Cofilin diffused throughout the cytosol of resting cells, but accumulated at the plasma membranes forming phagocytic vesicles in activated cells. (ii) At 1 microM, OA had little effect on the OZ-evoked translocation of cofilin, whereas 5 microM OA suppressed it completely. (iii) OA alone, which could not trigger the phagocytic respiratory burst, did not cause any change in the distribution of cofilin at such concentrations. Furthermore, in a superoxide-producing cell-free system employing membranous and cytosolic fractions, affinity-purified anti-cofilin antibody showed an enhancing effect. These results suggest that cofilin participates in the superoxide production of the OZ-activated phagocytes through dephosphorylation and translocation. The roles of cofilin in the activated leukocytes will be discussed.

Fetal cells in the maternal circulation: detection of Y-sequence by gene amplification.

Detection of a Y-specific sequence in the maternal circulation has clinical importance because it would be useful in determining fetal gender in mothers with severe X-linked disorders. The method described in this paper has the advantage of requiring only small amounts of maternal blood. Numerous attempts have been made to identify XY cells in the blood of mothers bearing male fetuses; however, the results have been controversial. In this study, a member of the DYZ1 family and the XY homologous region of the amelogenin gene were used as targets for polymerase chain reaction detection of the Y chromosome. The subjects in this study were a group of 100 pregnant women at 17-20 weeks' gestation and 30 puerperal women who had given birth 2-5 days previously. All of the former underwent amniocentesis, with venous blood samples drawn before the procedure. Forty-five fetuses were confirmed as male by karyotyping amniocytes, and 30 of these were positive for the Y sequence in the DYZ1 region (sensitivity 66.7%). However, ten of the 55 cases diagnosed as female were also positive, giving a specificity of only 81.8%. Thus, the positive and negative predictive values were each 75%. In the amelogenin gene study, a positive Y signal was not detected in any of the cases examined. This study demonstrates the usefulness of polymerase chain reaction detection of Y-specific sequences in the maternal circulation. However, further investigation is necessary to increase the reliability for clinical application, because the method does produce false-positive results.

Bisphosphonates for steroid induced osteoporosis.

OBJECTIVES: To assess the effects of bisphosphonates for the prevention and treatment of corticosteroid-induced osteoporosis. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group trials register, Medline up to 1997 and Embase1988-1997), and selected hand searching of reference lists was conducted. Hand searching of scientific abstracts from relevant meetings for the last five years was also done. An electronic search in Current Contents was done for the last six months. The Cochrane Controlled Trials Register (CCTR) will be searched for future updates. All languages were included in the search. For practical reasons only those in English were included, but all languages will be retrieved and translated for future updates. SELECTION CRITERIA: All controlled clinical trials (CCTs) dealing with prevention or treatment of corticosteroid-induced osteoporosis with bisphosphonates of any type and reporting the outcomes of interest were assessed. Trials had to involve adults only, and subjects had to be taking a mean steroid dose of 7.5 mg/day or more. DATA COLLECTION AND ANALYSIS: All data extraction was performed by two independent reviewers. Outcomes of interest included change in bone mineral density (BMD) at the lumbar spine and femoral neck at six and 12 months. If present, data on number of new fractures and withdrawals due to adverse effects were also extracted. All data extraction was performed by two independent reviewers. Both continuous and dichotomous data were analyzed using fixed effects models. When significant heterogeneity was present, a random effects model was used. MAIN RESULTS: A total of 13 trials, including 842 patients are included in this meta-analysis. Results are reported as a weighted mean difference of the percent change in BMD between the treatment and placebo groups, with trials being weighted by the inverse of their variance. The 95% confidence intervals (95% CI) are presented. At the lumbar spine, the weighted mean difference of BMD between the treatment and placebo groups was 4.3% (95% CI 2.7, 5.9). At the femoral neck, the weighted mean difference was 2.1% (95%CI 0. 01, 3.8). Although there was a 24% reduction in odds of spinal fractures [OR 0.76 (95%CI 0.37, 1.53)], this result was not statistically significant. REVIEWER'S CONCLUSIONS: Bisphosphonates are effective at preventing and treating corticosteroid-induced bone loss at the lumbar spine and femoral neck. Efficacy regarding fracture prevention cannot be concluded from this analysis, although bone density changes are correlated with fracture risk.

The Canadian SCORE questionnaire: optimizing the use of technology for low bone density assessment. Simple Calculated Osteoporosis Risk Estimate.

The Simple Calculated Osteoporosis Risk Estimation (SCORE) questionnaire is a tool to assist physicians to identify women who might require bone densitometry. The purpose of this study was to develop a Canadian SCORE and to assess validity and reliability. Twenty sites enrolled 307 postmenopausal women ages 50-70 yr. SCORE results were compared to hip and lumbar spine bone density assessed by dual X-ray absorptiometry. Sensitivity and specificity of a range of SCORE cut-points were assessed in a receiver operating characteristics analysis to determine the optimal cut-point for SCORE. With low bone density defined as a T-score < or = -2.0, a SCORE cut-point of 6 in women ages 50-59 yr displayed a sensitivity of 0. 96, 95% confidence interval (CI) (0.89, 1.00), a specificity of 0.51, 95% CI (0.43, 0.58). In women ages 60-70 yr, a SCORE cut-point of 8 displayed a sensitivity of 0.90, 95% CI (0.80, 0.97) and a specificity of 0.20, 95% CI (0.11, 0.29). The test-retest reliability (intraclass correlation coefficient) was 0.95. SCORE performed better in women in their fifties than women in ther sixties. Older women require higher SCORE cut-points. The use of SCORE as an initial measure for identifying those at risk for osteoporosis may reduce costs by limiting unnecessary tests.

Effect of cerebral venous congestion on the pressure-volume index in the evaluation of intracranial pressure dynamics.

Translocation of cerebrospinal fluid (CSF) between the intracranial and spinal subarachnoid space was blocked by ligating the cervical spinal core in eight cats under pentobarbital and nitrous oxide anesthesia, and the effects of cerebral venous congestion on the pressure-volume index (PVI), a measure relating the change in intracranial volume, and the logarithm of intracranial pressure (ICP) were evaluated. The changes in the volume-pressure response (VPR), a measure of intracranial elastance, were calculated simultaneously. Cerebral venous congestion was induced by lowering the head relative to the level of the heart by tilting the trunk of the animals to 20 degrees below horizontal. The presence of venous congestion was confirmed by an increase in the sagittal sinus pressure (SSP). The body position was shifted from horizontal prone (H1 group) to head-down tilt (D1 group) in four animals (group 1) and from head-down tilt (D2 group) to horizontal prone (H2 group) in the other four animals (group 2), and PVI and VPR were determined in each group. The changes in ICP and SSP with change of body position in group 1 were not significantly different from those in group 2, with both pressures changing by 7-8 mm Hg. PVI showed no significant differences between the H1 group and H2 group or between the D1 group and D2 group. The mean (+/- SEM) PVI for all measurements in the head-down tilt position (D1 and D2 groups) was significantly higher (0.50 +/- 0.02 ml; p < 0.01) than in the horizontal position (H1 and H2 groups; 0.35 +/- 0.02 ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Measurement of cytokine levels by coronary sinus blood sampling during cardiac surgery with cardiopulmonary bypass.

Interleukin-6, interleukin-8, and polymorphonuclear leukocyte elastase levels in coronary sinus blood were measured and compared with those in arterial blood drawn from the radial artery before and immediately after cardiopulmonary bypass (CPB) during coronary artery bypass grafting in 20 patients. We introduced coronary sinus blood sampling as a useful method for evaluation of myocardial metabolism, myocardial protection, and reperfusion injury during CPB, especially by measurement of cytokines. Because interleukin-6, interleukin-8, and polymorphonuclear leukocyte elastase are inflammatory mediators, we speculated that they might show higher levels in coronary sinus blood than in systemic arterial blood. The results obtained from the 20 patients showed that levels of interleukin-6, interleukin-8, and polymorphonuclear leukocyte elastase increased immediately after CPB (p < .01), but there was no significant difference in these levels between coronary sinus and systemic arterial blood. We conclude that the myocardium is not a predominant source of their release during CPB. Our results also showed that the measurement of these cytokines in systemic arterial blood reflected their levels in the whole body, including the myocardium, even during cardiac operation with CPB.

99mTc-HMDP accumulation in a phyllodes tumor of the breast, a case report.

In a patient with breast tumor, 99mTc-HMDP accumulation in the tumor was recognized in evaluating bone metastasis. Surgery and histopathology revealed that the tumor was a phyllodes tumor. This scintigraphic finding was thought to be rare and phyllodes tumor should be included in differential diagnosis when 99mTc-HMDP accumulation in the breast was recognized.

Detection of selenium in generalized and localized argyria: report of four cases with X-ray microanalysis.

Electron microscopic and X-ray microanalytic studies were performed on four cases of argyria; one generalized and three localized. Deposition of electron dense granules was predominantly found on elastic fibers and around basal laminas of secretory portions of eccrine glands, although the amount of deposition was much less in the case of generalized argyria. In all four cases, X-ray microanalysis revealed that the depositions consisted mainly of silver, selenium, and sulfur. The importance of selenium in the detoxification of heavy metals was discussed.

Incident-angle dependence of the sensitivity of second-harmonic generation of an oxacyanine dye in an LB film.

The second-harmonic generation of a cyanine dye in an LB film was investigated by varying the incident angle of the excitation laser. There was a clear dependence on the incident angle, which was simulated by a simple model. Thus, a highly sensitive determination should be carried out at such an angle where the efficiency of the second-harmonic generation shows its maximum.

[Assessment of myocardial viability with 201Tl myocardial SPECT 24 hours after injection].

Recently many investigators reported that conventional stress-redistribution myocardial scintigraphy with 201Tl underestimated the presence of ischemic but viable myocardium. We studied the usefulness of 24 hour 201Tl myocardial single photon emission computed tomography (SPECT) to assess myocardial viability and investigated the factors affect to the quality of 24 hour SPECT images. Study patients were consisted with 70 patients with old myocardial infarction (OMI), 72 patients with angina pectoris without OMI (AP) and 43 patients with angiographically proven normal coronary arteries. To obtain SPECT images, 10 minute and 4 hour imagings were sampled 30 seconds per projection. Twenty-four hour imaging was sampled 60 seconds per projection. Twenty-four hour images were visually interpreted as good, moderate and poor quality. Then study patients were divided into 2 groups, group A with good 24 hour images and group B with moderate or poor 24 hour images. One hundred and fifty-eight patients (85.4%) of study patients had 24 hour SPECT images on a good quality. Only 4 patients (2.2%) had poor quality SPECT. All of these 4 patients had broad myocardial infarction. In patients with OMI 61 patients (87.1%), in AP 63 patients (87.5%) and in normal 35 patients (81.4%) had a good 24 hour SPECT. Total sampling counts and myocardial ROI counts were significantly higher in group A than in group B. Body weight was significantly higher and there were more male patients in group B than in group A. Late redistribution was seen in 20 patients (28.5%) with OMI and in 11 patients (15.3%) with AP.(ABSTRACT TRUNCATED AT 250 WORDS)

[Changes in the intracranial pressure dynamics with head-down tilt in cats].

To determine changes in the intracranial pressure dynamics under the condition of head-down tilt, the pressure-volume relationship in the cranial cavity as well as changes in intracranial pressure were studied experimentally in 31 cats. The animals were divided into three groups; with the horizontal, with the head-down and one with the head-up position. In each group the pressure volume index (PVI) and cranial cavity compliance (C) were examined under controlled respiration. The following results were obtained. (1) The values of PVI and C were significantly greater in the head-down tilt group as compared to those in the other groups, in spite of the increase in intracranial pressure level. (2) In the situations of craniospinal blockade by epidural ligation of spinal cord at C2 level, the PVI and C were also higher in the head-down tilt group than in the horizontal group. These results indicate that the pressure buffering capacity is increased with head-down tilt with respect to intracranial volume loading. It appears that the buffering mechanism does not depend on movement of the cerebrospinal fluid, but on the increased cerebral venous vascular bed with the communication between extracranium and intracranium.

[Mathematical simulation of intracranial condition--Part 1. Linear model stimulation].

We developed a linear mathematical model of the intracranial vessels, which reflects changes of the pulse wave (pulse pressure) of intracranial pressure after ligation of the internal jugular vein. The model composed of eight major variables: 1. resistance of arteries, 2. resistance of small arteries and capillary vessels, 3. resistance of veins, 4. resistance of internal jugular and vertebral veins, 5. compliance of arteries, 6. compliance of small arteries and capillary vessels, 7. compliance of veins and 8. intracranial compliance. All variables are presumed to have linear elements and replaced with electrical elements. The model of neck dissection is expressed as the change of resistance of the internal jugular and vertebral veins. Intracranial condition is expressed as the pulse wave (pulse pressure) of intracranial pressure and driving pressure. After unilateral ligation of the internal jugular vein, the pulse wave of intracranial pressure increased 24% and, after bilateral ligation of the internal jugular vein, it increased 55%. After unilateral ligation of the internal jugular vein, the pulse wave of intracranial pressure increased 27%, and, after bilateral ligation, it increased 79%. When intracranial compliance is normal, the respective ratios of pulse wave of intracranial pressure and driving pressure to cross-sectional area decreased, whereas those after increase of intracranial compliance increased.