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Background: Allergen immunotherapy (AIT) is the only disease-modifying treatment for immunoglobulin (Ig) E-mediated allergy. Owing to the high prevalence and early onset of hay fever and pollen-food allergy syndrome (PFAS), a safer and simpler treatment method than conventional AIT is needed. To develop a local nasal immunotherapy using an ointment containing hypoallergenic pollen and assess its efficacy in mice and healthy humans., Methods: Hypoallergenicity was achieved by combining pollen and galactomannan through the Maillard reaction to create birch pollen-galactomannan conjugate (BP-GMC). The binding of galactomannan to Bet v 1 was confirmed using electrophoresis and Western blotting (WB). Binding of specific IgE antibodies to BP-GMC was verified using enzyme-linked immunosorbent assay (ELISA) and basophil activation test (BAT). The localization of BP-GMC absorption was confirmed using a BALB/c mouse model. BP-GMC mixed with white petrolatum was intranasally administered to 10 healthy individuals (active drugs, 8; placebo, 2) for 14 days., Results: In electrophoresis and WB, no 17-kDa band was observed. In ELISA and BAT, BP-GMC did not react to specific IgE but was bound to IgA and IgG. In the mouse model, BP-GMC was detected in nasopharyngeal-associated lymphoid tissues. In the active drug group, the salivary-specific IgA level significantly increased on day 15 (p = 0.0299), while the serum-specific IgG level significantly increased on day 85 (p = 0.0006)., Conclusions: The BP-GMC ointment rapidly produced antagonistic antibodies against IgE; it is safe and easy to use and might serve as a therapeutic antigen for hay fever and PFAS., (Copyright © 2023 Japanese Society of Allergology. Published by Elsevier B.V. All rights reserved.)

Fuligocandin B ( 2 ) is a novel natural product that can overcome TRAIL resistance. We synthesized enatiomerically pure fuligocandin B ( 2 ) and its derivative 5'-I fuligocandin B ( 4 ), and found that the latter had an improved biological activity against the human gastric cancer cell line, AGS. We attached a biotin linker and photoactivatable aryl diazirine group to 5'-I fuligocandin B ( 4 ), and employed a pull-down assay to identify valosin-containing protein (VCP/p97), an AAA ATPase, as a 5'-I fuligocandin B ( 4 ) target protein. Knock-down of VCP by siRNA enhanced sensitivity to TRAIL in AGS cells. In addition, 4 enhanced CHOP and DR5 protein expression, and overall intracellular levels of ubiquitinated protein. These data suggest that endoplasmic reticulum stress caused through VCP inhibition by 4 increases CHOP-mediated DR5 up-regulation, which enhances TRAIL-induced cell death in AGS cells. To the best of our knowledge, this is the first example to show a relationship between VCP and TRAIL-resistance-overcoming activity in cancer cells.

Cell migration plays important roles in embryonic development and inflammation, and this process is highly regulated to ensure tissue homeostasis. A number of barriers exist to prevent the inappropriate migration of leukocytes into healthy peripheral tissues, including retention of these cells in the inactive state and maintenance of the integrity and charge of the vascular endothelium. However, active signals also are likely to exist that can repulse cells or abolish existing cell migration. One such paradigm exists in the developing nervous system, where neuronal migration is mediated by a balance between chemoattractive and chemorepulsive signals. The ability of the guidance molecule netrin-1 to repulse or abolish attraction of neuronal cells expressing the UNC5b receptor makes it an attractive candidate for the regulation of inflammatory cell migration. Here, we show that netrin-1 is expressed on vascular endothelium, where it is regulated by infection and inflammatory cytokines. The netrin-1 receptor UNC5b is strongly expressed by leukocytes, upon which netrin-1 acts as a potent inhibitor of migration to different chemotactic stimuli both in vivo and in vitro. These data suggest that endothelial expression of netrin-1 may inhibit basal cell migration into tissues and that its down-regulation with the onset of sepsis/inflammation may facilitate leukocyte recruitment.

Retinoic acid (RA) is an important coordinator of mammalian organogenesis. RA is implicated in critical lung developmental events. Cell proliferation is precisely regulated during development. We investigated the effect of RA on proliferating mesenchymal cells in both whole organ lung cultures and cell cultures. The potential pathways required for the response were studied in cultures of lung mesenchymal cells from embryonic day (e) 12. We observed an RA-dependent reduction in proliferation of mesenchymal cells in both whole organ and in cell culture. In mesenchymal cell cultures, RA decreased proliferation in lung mesenchymal cells by 72%. This was associated with a decrease of erk-1/2 activity by 68%. Mesenchymal cell proliferation is erk-1/2 dependent. Erk-1/2 can be activated by G-protein coupled receptors (GPCR) or tyrosine kinase receptors (RTK). RA treatment altered both the RTK and the GPCR pathways in primary lung mesenchymal cells. The Epidermal Growth Factor (EGF) dependent erk-1/2 activation was increased by 35% whereas the G(i)-protein cascade was inhibited by 44% in cells treated with RA. Our results suggest that RA decreases proliferation of lung mesenchyme via a G(i)-protein and the erk-1/2 signaling cascade.

The ligand Netrin-1 and its receptors DCC and UNC5H2 are critical for the regulation of neuronal migration in nervous system development. Here we demonstrate expression of these molecules in lung development. The mRNA expression profiles of Netrin-1, DCC and UNC5H2 are developmentally regulated during embryonic mouse lung formation. Netrin-1 shows a bimodal expression pattern with elevated mRNA levels early followed by a second peak in late gestation. Peak expression of DCC occurs early in development whereas expression of UNC5H2 peaks late in development. We also demonstrate localization of Netrin-1, DCC and UNC5H2 during the stages of lung development. We present evidence that these proteins are modulated spatially in the mesenchyme and epithelium during lung organogenesis.

First described as an axonal guidance cue through its repulsive effect on neurons expressing its receptor Roundabout (Robo), the Slit ligand has effects on cell migration, axon branching and elongation. Indirect evidence implicates Slit and Robo in lung development. We now demonstrate that Slit-2 and Slit-3 are developmentally regulated in embryonic murine lung. Immunohistochemistry demonstrates Slit-2 and Slit-3 expression by the pulmonary mesenchyme and airway epithelium. Robo-1 and Robo-2 are also expressed by the developing mesenchyme and airway epithelium. As lung development progresses, Robo-1 and Robo-2 expression localizes to only the airway epithelium. We conclude Slit/Robo are expressed in temporo-spatially adjacent domains suggesting interactive roles in pulmonary bronchiolar development., (Copyright 2003 Elsevier Science B.V.)

The G protein, G(ialpha(2)), regulates a number of cellular functions including cell migration, proliferation, and differentiation. The transduction of signal depends on the ability of the alpha subunit to cycle between a GDP bound and an active GTP bound state capable of interacting with intracellular enzymes. Here, we now report the novel interaction of gip2 (constitutively activated G(ialpha(2))) with the cytoplasmic domain of UNC5H2. Like G(ialpha(2)), we found that UNC5H2 is widely expressed particularly in cells which migrate. UNC5H2 binds G(ialpha(2)) when it is charged with GTP. The interaction of G(ialpha(2)) and UNC5H2 liberated adenylyl cyclase from G(ialpha(2)) inhibition. Thus, by sequestering the alpha subunit, UNC5H2 is a novel inhibitor of G(ialpha(2)) thereby increasing intracellular cAMP levels. The expression of UNC5H2 in the brain and immune system suggests that this novel inhibitor of G protein signaling may have broad significance for axonal guidance and chemotaxis.

Somatostatin is a neurotransmitter with diverse effects including anti-proliferation in a wide range of normal and neoplastic cells, and occasionally growth stimulatory and neurotrophic actions. Stress-activated protein kinase or c-Jun N-terminal kinase (SAPK/JNK) can also induce growth arrest and occasionally growth stimulation. However, the relationship between somatostatin and SAPK/JNK is less clear. Here we report that the binding of somatostatin to the somatostatin receptor type V (SSTR5) upregulates SAPK/JNK activity. We also show that this activation is mediated by Galpha(12) and Galpha(13). This study demonstrates that SSTR5 is the heptahelical receptor that activates SAPK/JNK via the G(12) family G proteins.

Heterotrimeric guanine nucleotide-binding (G) proteins transduce a wide variety of receptor-mediated signals to effectors that are involved in numerous cellular functions, including cell proliferation and differentiation. Thrombin and bombesin/gastrin-releasing peptide mediate their effects via G protein-coupled receptors to regulate lung growth and development. The growth responses of these ligands are likely to be mediated via the Gi subfamily of G proteins, specifically via Galphai2. We hypothesized that Galphai2 is expressed in the lung during ontogeny in a growth-dependent manner, and that Galphai2 regulates cell growth. We demonstrate that Galphai2 is present in the developing lung of Sprague-Dawley rats, and that its expression is enhanced between embryonic Day 19 and postnatal Day 2. The strongest expression occurs in the fetal airway epithelium, and this expression in fetal airway cells is growth-dependent. Galphai2 is localized to the plasma membrane, a location consistent with interaction with growth factor receptors. Inhibition of Gi-family signal transduction by pertussis toxin (10 ng/ml) inhibits DNA synthesis in embryonic Day 19 in fetal airway epithelium. Galphai2 is likely to be a key mediator of growth signals in the developing lung.

Missense mutations in the 695-amino acid form of the amyloid precursor protein (APP695) cosegregate with disease phenotype in families with dominantly inherited Alzheimer's disease. These mutations convert valine at position 642 to isoleucine, phenylalanine, or glycine. Expression of these mutant proteins, but not of normal APP695, was shown to induce nucleosomal DNA fragmentation in neuronal cells. Induction of DNA fragmentation required the cytoplasmic domain of the mutants and appeared to be mediated by heterotrimeric guanosine triphosphate-binding proteins (G proteins).

The clinical usefulness of cefpodoxime proxetil (CPDX-PR) was investigated in the treatment of pneumonia and chronic airway infections occurring in patients first visiting our outpatient clinic or those being treated at the outpatient clinic. CPDX-PR was orally administered twice a day after meals at a dose of 100-200 mg for acute respiratory tract infections and at a dose of 200 mg for chronic respiratory tract infections. Excellent, good, fair, and poor responses were observed in 20, 33, 10, and 3 of 66 patients (4 with acute bronchitis, 27 with pneumonia, and 35 with acute exacerbation of chronic airway infection), respectively, demonstrating an 80.3% efficacy rate (53/66). Causative organisms, including Streptococcus pneumoniae, were all eradicated from the patients whose causative organisms were examined over time, although 2 of the patients were superinfected with Pseudomonas aeruginosa. There were no serious adverse reactions or abnormal changes in laboratory test results. It was concluded that CPDX-PR could be used as a first-choice drug for the treatment of respiratory tract infections at an outpatient clinic, and that this drug should acquire greater importance in particular consideration of recent increases in infections with S. pneumoniae.

A case of vulvar leiomyoma with extensive myxoid change in a 40 year old female is described. The tumor had a unique connection with a non-degenerative leiomyoma that compressed the rectum and the bladder. Scattered smooth muscle cells in a loose myxoid stroma were immunoreactive for desmin. Fibroblast-like spindle cells were immunoreactive for vimentin but not for desmin. The initial, although incorrect, pathological diagnosis of the tumor was aggressive angiomyxoma based on the similarity in both clinical and pathological aspects with this more invasive tumor. Myxoid vulvar leiomyoma should also be differentiated from angiomyofibroblastoma. The key to the differential diagnosis is the presence of interlacing smooth muscle cells and an awareness of tendency toward myxoid change in vulvar leiomyomas.

Nineteen-hydroxyandrostenedione (19-OHA) is considered to be an obligatory intermediate of estrogen synthesis. To clarify the role of 19-OHA in the human ovary, the following experiments were undertaken. Ovarian and peripheral vein blood in mid follicular and luteal phase were obtained from 14 women during gynecological surgery. The concentrations of androstenedione (A), testosterone (T), 19-OHA, estrone (E1) and estradiol (E2) were measured by gas chromatography mass spectrometry using deuterium-labeled steroids as internal standard. The effect of human chorionic gonadotropin (hCG) on those steroids was also studied in mid luteal phase. The concentrations of 19-OHA in ovarian vein were 1.20 +/- 0.16 ng/ml (mean +/- SE) in mid follicular and 0.78 +/- 0.18 ng/ml in mid luteal phase. The steroid levels measured in ovarian vein were significantly higher than those in peripheral vein. Though the administration of hCG in mid luteal phase enhanced A and E2 levels in ovarian vein, the levels of T, 19-OHA and E1 were not altered. Significant correlation between the levels of E2 and A was observed while none of the steroids correlated with 19-OHA. These results suggest, for the first time, that 19-OHA is produced and secreted from the human ovary. The question as to the physiological and biosynthetic role of ovarian 19-OHA has to be answered in the future.

Ten patients (8 with moderate or severe infections, 2 for prevention of postoperative infections) were treated with cefoxitin. The patients were given cefoxitin of 2 approximately 9 g, given once, or in 2 approximately 3 divided doses. Clinical efficacy was good in 7 cases, fair in 2 and poor in 1. Transient nausea and vomiting occurred in 1 patient when a direct intrauterine injection (1 g of cefoxitin) was given, however, no side effects were observed in the other patients. The result of this study demonstrates that cefoxitin when given massively is effective in achieving bacteriologic and clinical cure in treatment of infections and for prevention of postoperative infections in the field of obstetrics and gynecology.

In order to study the metabolic fate of circulating pregnenolone sulfate (P5S) during pregnancy, 30 mg of newly synthesized tetra (2,2,4,6)-deuterated P5S was administered to the maternal vein 60 minutes prior to cesarean section at term. All pregnant women were volunteers and had been informed of nature of this study. The placenta, maternal urine and blood samples from maternal vein (MV), umbilical cord (U) were collected and deuterated metabolites were analysed by gas chromatography mass spectrometry with a multiple ion detector. Total amounts of metabolites were measured and the ratio of deuterated steroid to the total amounts were calculated (d%). d% of P5S, 16 alpha OH-P5S, 17 alpha OH-P5S, 20 alpha-dihydro-P5S (20P5S), 20P5 and progesterone (P4) in MV were 84.5, 51.6, 95.5, 85.1, 71.2 and 10.9%, respectively. In the placental tissue, 20P5, 20P4 and P4 were also found and d% of these steroids were calculated as 16.1, 3.2 and 3.1%. Only P4 was found with d% of 11.2% in U. In the urine collected for 2 hours after deuterated P5S administration, P5S (40.6%), 20P5S (56.6%) and pregnanediol (34.8%) were identified. Deuterated C19, C18 steroids were not detected in any of the samples studied. When 30 mg of non labeled P5S was also administered to MV at term, the levels of P5S, 20P5S, P4 and 20P4 in MV rose, but the levels of DHA-S were not changed. These results indicate that the circulating P5S in MV can be a precursor of 20P5S, 17P5S, 16P5S, 20P4 and P4, but can not be the precursor of C19, C18 steroid.

To establish an efficient rearing system for weaned cynomolgus infants, 51 groups composed of 4 weanlings each were formed. Of them each of thirty-eight groups had an adult female "nurse" monkey, who had no kinship with any of the 4 weanlings. The nurse was present during the first 4 weeks after weaning. The daily incidence of watery diarrhea was analyzed to assess the usefulness of the nurse monkey for stabilizing physiological conditions of the weanlings. Diarrheal incidence was significantly lower in the 38 groups with a nurse than in the 13 control groups without a nurse throughout the 4-week period of observation. This trend continued for more than 5 weeks after removal of the nurse. Thus, the time and manpower necessary for treating infant diarrhea were greatly reduced in this rearing system.