Factors affecting guardians' decision-making regarding the SARS-CoV-2 vaccine.

BACKGROUND: In Japan, the vaccination rate against the SARS-CoV-2 vaccine for children was low. Therefore, in this study we investigated the factors influencing guardians' decision-making regarding vaccination of their children. METHODS: From November 1, 2022 to March 31, 2023, pediatric clinics, departments, and midwifery clinics in Saitama Prefecture requested guardians of children under the age of 15 to complete an online questionnaire. RESULTS: Responses were obtained from 894 guardians of children aged 6 months to 15 years; 142 had had one of their children vaccinated at least once and 629 had not had any of their children vaccinated. Among guardians who had not had any of their children vaccinated, "the Age of children" was significantly younger (p < 0.001) and "Prevalence" (p < 0.001), "Free vaccination" (p < 0.001), and "Intentions of national and local governments" (p = 0.005) were selected as reasons significantly less frequently in comparison to guardians who had vaccinated their children. "Japanese adverse reactions" (p < 0.001), "Japanese effectiveness" (p < 0.001), "Adverse reactions" (p < 0.001), "History of adverse reactions" (p < 0.001), and "Reputation of friends" (p = 0.006) were selected significantly more frequently by guardians who had not had any of their children vaccinated. CONCLUSIONS: Guardians who had had one of their children vaccinated at least once emphasized the importance of prevalence and free vaccination. On the other hand, guardians who had not had any of their children vaccinated placed particular importance on adverse reactions and the Japanese data on effectiveness. To guide the decision-making of guardians, it is necessary to quickly collect and publish data on adverse reactions and effectiveness, particularly in Japanese individuals, so that citizens can decide whether to vaccinate themselves and their children.

Molecular cloning, molecular evolution and gene expression of cDNAs encoding thyrotropin-releasing hormone receptor subtypes in a teleost, the sockeye salmon (Oncorhynchus nerka).

Molecular cloning of thyrotropin-releasing hormone receptors (TRHR) was performed in a teleost, the sockeye salmon (Oncorhynchus nerka). Four different TRHR cDNAs were cloned and named TRHR1, TRHR2a, TRHR2b and TRHR3 based on their similarity to known TRHR subtypes in vertebrates. Important residues for TRH binding were conserved in deduced amino acid sequences of the three TRHR subtypes except for the TRHR2b. Seven transmembrane domains were predicted for TRHR1, TRHR2a and TRHR3 proteins but only five for TRHR2b which appears to be truncated. In silico database analysis identified putative TRHR sequences including invertebrate TRHR and reptilian, avian and mammalian TRHR3. Phylogenetic analyses predicted the molecular evolution of TRHR in vertebrates: from the common ancestral TRHR (i.e. invertebrate TRHR), the TRHR2 subtype diverged first and then TRHR1 and TRHR3 diverged. Reverse transcription-polymerase chain reaction analyses revealed TRHR1 transcripts in the brain (hypothalamus), retina, pituitary gland and large intestine; TRHR2a in the brain (telencephalon and hypothalamus); and TRHR3 in the brain (olfactory bulbs) and retina.

Eosinophils are required for the induction of bronchial hyperresponsiveness in a Th transfer model of BALB/c background.

BACKGROUND: Helper T (Th) cells are deeply involved in the pathophysiology of bronchial asthma, such as eosinophilic inflammation, bronchial hyperresponsiveness (BHR), airflow limitation and remodeling. It is still unclear whether Th cells contribute to BHR independently of eosinophilic inflammation. The double GATA (dblGATA) site is a high-affinity GATA-binding site in the GATA-1 promoter. dblGATA site-deficient (Delta dblGATA) mice lack eosinophils. METHOD: Ovalbumin (OVA)-reactive Th clones were transferred into Delta dblGATA and wild-type (WT) mice of BALB/c background. The number of eosinophils in the bronchoalveolar lavage fluid (BALF) and bronchial responsiveness to methacholine were examined after OVA challenge. RESULTS: The number of BALF eosinophils was significantly increased in WT mice, but not detectable in Delta dblGATA mice. BHR was also induced in WT mice, but significantly attenuated in Delta dblGATA mice. CONCLUSION: Eosinophils are involved in T-cell-mediated BHR.

Murine Th clones confer late asthmatic response upon antigen challenge.

BACKGROUND: Helper T (Th) cells are deeply involved in the pathophysiology of bronchial asthma, such as eosinophilic inflammation, bronchial hyperresponsiveness and remodeling. However, it is still unclear how Th cells contribute to airflow limitation, another cardinal feature of bronchial asthma. METHOD: Unprimed BALB/c mice were transferred with ovalbumin (OVA)-reactive Th clones. Pulmonary function was monitored using a Buxco BioSystem Plethysmograph before and after OVA challenge. RESULTS: When Th-transferred mice were challenged with OVA, enhanced pause (P(enh)), an indicator of airflow limitation was significantly increased 6 and 24 h after challenge, while no response was observed 30 min after challenge. Neither bovine serum albumin, an irrelevant antigen, challenge on Th-transferred mice nor OVA challenge on Th-non-transferred mice caused airway responses. CONCLUSION: Th cells conferred antigen-induced airflow limitation to unprimed mice.

Evaluation of cysteinyl leukotriene-induced contraction of human cultured bronchial smooth muscle cells.

BACKGROUND: Cysteinyl leukotrienes (cysLTs) are major mediators involved in bronchial asthma, particularly bronchial constriction. However, a contractile response of human bronchial smooth muscle cells (hBSMCs) to cysLTs has not been well characterized at cellular level. METHODS: A contraction assay using collagen gel embedded with cultured hBSMCs was established to analyze contractile responses at cellular level. Contractile responses to several constrictors including cysLTs were evaluated in 6-day cultured gels with varying fetal bovine serum (FBS) concentrations. RESULTS: Removal of FBS from the culture for the designated time periods resulted in increased contractile responses. CysLT-induced contraction of the gel became more pronounced and reproducible. CysLT(1)R expression on the hBSMCs was significantly increased by the removal of FBS. CONCLUSION: Contractile responses of hBSMCs to cysLTs can be evaluated at cellular level for the first time. This experimental system may be useful for the in vitro evaluation and future development of cysLTs antagonists.

A contraction assay system using established human bronchial smooth muscle cells.

BACKGROUND: To further understand the mechanisms of airway obstruction in asthma, it is crucial to investigate contractile responses of human airway smooth muscle. An in vitro assay system employing collagen gels embedded with well-established bronchial smooth muscle cells of human origin was explored in the present study. METHODS: Commercially available cultured human bronchial smooth muscle cells were embedded into a collagen gel. Well-known constrictors, histamine and methacholine, were added to the gel. The gel images were captured by an image analyzer, and contractile responses were evaluated. RESULTS: Histamine and methacholine induced contraction of the gels in a dose-dependent manner. Pyrilamine, an H1 receptor antagonist, inhibited gel contraction in an agonist-specific manner. CONCLUSION: Our contraction assay system, employing widely distributed cultured cells, was highly reproducible and precise. It may go a long way toward understanding mechanisms of asthmatic responses and evaluation of antiasthma drugs.

Airway eosinophilic inflammation is attenuated in conserved noncoding sequence-1-deficient mice.

BACKGROUND: Conserved noncoding sequence-1 (CNS-1) is an important regulatory element for T helper 2 cytokine expression. IL-4, IL-5 and IL-13 expression as well as serum IgE level were attenuated in CNS-1-/- mice. METHOD: CNS-1-/- and CNS-1+/+ mice were sensitized with ovalbumin (OVA) followed by antigen challenge. The number of eosinophils and T helper 2 cytokine concentration in the bronchoalveolar lavage fluid, OVA-specific IgE antibody (Ab) in the serum and bronchial responsiveness to methacholine were examined. RESULTS: Bronchoalveolar lavage fluid eosinophilia was significantly attenuated in CNS-1-/- mice compared to CNS-1+/+ mice, which were sensitized with OVA/aluminum once. OVA-specific IgE Ab was also attenuated. When mice were sensitized with OVA/aluminum twice, induction of eosinophilia and OVA-specific IgE Ab was not significantly different between CNS-1-/- and CNS-1+/+ mice. CONCLUSION: CNS-1 locus regulates eosinophilic inflammation in vivo.

Synthesis of FF8181-A.

Both enantiomers of FF8181-A were synthesized through optical resolution from the known Diels-Alder reaction product in 15 steps. The absolute configuration of the natural product was determined to be 1S,5S,5aS,9aS,9bS.

Character impact odorants of high-heat skim milk powder isolated by simultaneous distillation-extraction.

To identify the character impact odorant of high-heat skim milk powder (HHSMP), a comparative study using ultrahigh-temperature (UHT) milk was performed. Aroma concentrate was prepared by column adsorption combined with simultaneous distillation-extraction. Aroma extract dilution analysis (AEDA) revealed 58 aroma peaks with flavor dilution (FD) factors ranging from 10 to 3000; from these, 41 compounds were identified and 7 compounds were tentatively identified (FD factor > or = 300). Among these HHSMP and UHT milk components, methyl 2-methyl-3-furyl disulfide and bis(2-methyl-3-furyl) disulfide, which appeared to be generated during the processing of each product, were identified. When the results of the AEDA of both samples were compared, it was considered that the characteristic aroma of HHSMP was not explained by a single compound but instead formed from a mixture of several types of compounds contained in common with the UHT milk. The contribution of these compounds to the aroma of HHSMP was confirmed by an aroma simulation experiment.

IL-2-induced IL-9 production by allergen-specific human helper T-cell clones.

BACKGROUND: IL-9 is an important cytokine in allergic diseases such as asthma, atopic dermatitis, etc. T helper (Th) cells seem to be the main source of IL-9. Cellular and molecular mechanisms of IL-9 production by human Th cells have been poorly understood. METHODS: Dermatophagoides farinae(Der f)-specific Th clones were established from peripheral blood lymphocytes of atopic asthmatics, and cytokine synthesis in response to various stimuli was determined by specific ELISAs. RESULTS: IL-9 was produced by 14 of 27 human Th clones upon T cell receptor (TCR) stimulation, immobilized anti-CD3 antibody (Ab). IL-9 production was significantly enhanced by the addition of anti-CD28 Ab into the culture, indicating the role of costimulatory signal on IL-9 synthesis. Pharmacologically, IL-9 production was induced by ionomycin (IOM) alone, and enhanced by phorbol 12-myristate 13-acetate (PMA). rIL-2 induced IL-9 production by 8 out of 19 Th clones. IL-9 production by Th clones stimulated with immobilized anti-CD3 Ab was significantly suppressed by the addition of anti-IL-2 neutralizing Ab into the culture. CONCLUSION: Approximately half of the Der f-specific Th clones derived from atopic asthmatics produced IL-9 upon TCR stimulation. Ca(2+) signal, CD28 signal, and IL-2 receptor signal seem to play important roles in IL-9 production by human Th cells. Moreover, synthesis of IL-9, a Th2 cytokine, is dependent on IL-2, a Th1 cytokine, which is produced by Th cells themselves.

IL-9 production by peripheral blood mononuclear cells of atopic asthmatics.

BACKGROUND: IL-9 might play a critical role in pathogenesis and development of atopic asthma, but there are few reports on allergen-specific IL-9 production by peripheral blood mononuclear cells (PBMCs) obtained from adult asthmatics. METHODS: PBMCs were obtained from adult atopic asthmatics and incubated with Dermatophagoides farinae(Der f) extract for the designated time periods. The resulting supernatants were assayed for IL-9 by specific sandwich ELISA. RESULTS: IL-9 production was detectable on day 2 and reached maximum on day 6 after stimulation of PBMCs with Der f extract. IL-9 production in response to Der f extract increased in a dose-dependent manner. CD2- or CD4-bearing cell depletion completely abolished IL-9 production by PBMCs, while CD8-bearing cell depletion did not affect it. CONCLUSION: CD4+ lymphocytes are the principal source of IL-9 produced by PBMCs of adult atopic asthmatics.

Effect of formoterol on allergen-induced cytokine synthesis by atopic asthmatics.

BACKGROUND: Effect of formoterol, long-acting beta(2 )agonists, on T cell cytokine synthesis was examined. METHODS: Human peripheral blood mononuclear cells were obtained from atopic asthmatics, and stimulated with Dermatophagoidesfarinae extract. Various concentrations of formoterol were added from the start of some cultures. Cytokine production and cell proliferation were analyzed. RESULTS: Allergen-induced IL-5, IL-13, and IFN-gamma production of PBMC were significantly suppressed by formoterol in a dose-dependent manner, whereas the proliferation response was not suppressed. CONCLUSION: Formoterol downregulates T cell functions of atopic asthmatics in vitro.

Lipid synthesis and acyl-CoA synthetase in developing rice seeds.

Developing rice seeds rapidly accumulated storage lipids between 5 and 12 d after flowering. The contents of palmitic, oleic, and linoleic acids increased throughout seed development, while the alpha-linolenic acid content remained low. The activity of acyl-CoA synthetase varied coincidentally during the period of lipid accumulation, and rice seeds had a sufficient capacity to supply acyl-CoA substrates for TAG synthesis. Acyl-CoA synthetase showed a broad specificity for native FA of rice seeds except for stearic acid, and pi electrons of a delta9-delta11 double bond in the C16-C18 acyl chains were required for its maximal activity.

IL-2-induced IL-13 production by allergen-specific human helper T cell clones.

BACKGROUND: Interleukin (IL)-5 and IL-13 are important cytokines in allergic diseases such as asthma and atopic dermatitis. We have reported that the production of IL-5 and IL-13 by mite-responsive T helper cells (Th) is controlled under similar signal requirements, but precise mechanisms are not yet well characterized. METHODS: Allergen-specific Th clones were established from peripheral blood lymphocytes of atopic asthmatics, and cytokine synthesis in response to various stimuli was determined by specific ELISAs. IL-13 gene expression was enumerated by quantitative real-time PCR. RESULTS: IL-13 production was clearly induced by IL-2. IL-13 mRNA expression was also induced. CONCLUSION: The IL-2 signal by itself causes IL-13 synthesis independent of T cell receptor stimulation.

Comparison of IL-17 production by helper T cells among atopic and nonatopic asthmatics and control subjects.

BACKGROUND: T cells, eosinophils, and neutrophils are strongly involved in the pathogenesis of bronchial asthma. Mechanisms that influence neutrophil accumulation and activation in asthma still remain relatively obscure. There is data indicating that IL-17 is produced by T cells and causes the release of neutrophil-mobilizing cytokines from airway epithelial cells, and that in this way it may regulate airway neutrophilia. METHODS: Peripheral blood mononuclear cells (PBMC) obtained from atopic asthmatics (AA), nonatopic asthmatics (NA), and normal control subjects (NC) were stimulated by immobilized anti-CD3 antibody (Ab) plus soluble anti-CD28 Ab or Dermatophagoidesfarinae (Df) extract. Df-reactive T cell clones were established from PBMC of AA and cultured in the presence of various stimulants. The resulting supernatants were assayed for IL-2, IL-4, IL-5, IL-13, IL-17, and IFN-gamma by specific ELISAs. RESULTS: PBMC obtained from AA, NA, and NC all produced IL-17 upon immobilized anti-CD3 Ab plus soluble anti-CD28 Ab stimulation. IL-17 production in response to Df extract was significantly induced only in AA. The amount of IL-17 produced by T cell clones stimulated with immobilized anti-CD3 Ab plus soluble anti-CD28 Ab was negatively, but only weakly, correlated with that of IL-4, but not correlated with IL-2, IL-5, IL-13, and IFN-gamma production. CONCLUSION: T cells producing IL-17 in response to Df antigen exist in the peripheral blood of the sensitized AA. IL-17 production might be regulated by unique mechanisms different from those governing Th1 versus Th2 differentiation.

Correlation of allergen-induced IL-5 and IL-13 production by peripheral blood T cells of asthma patients.

BACKGROUND: Helper T cells and T cell cytokines are implicated in allergic disorders such as asthma and atopic dermatitis (AD). We reported enhanced interleukin-5 (IL-5) production by peripheral blood T cells of asthmatic patients. Production of cytokines, particularly IL-5 and IL-13, by peripheral blood mononuclear cells (PBMC) obtained from bronchial asthma (BA) and AD patients was investigated in the present study. METHODS: PBMC were cultured in the presence of either polyclonal activator, phorbol ester plus Ca2+ ionophore or Dermatophagoides farinae (Df) antigen. The resulting supernatants were assayed for IL-4, IL-5, and IL-13 by specific ELISAs. RESULTS: IL-5 and IL-13 production in response to Df antigen is significantly higher in allergic groups compared to control subjects. IL-5 responses induced by Df antigen were strongly correlated with IL-13 responses. CONCLUSION: In spite of IL-13 gene proximity to IL-4 gene, IL-5 production was more strongly correlated to IL-13 production in the BA group, suggesting a common control mechanism that regulates the IL-5 and IL-13 gene.