Topical application of activator protein-1 inhibitor T-5224 suppresses inflammation and improves skin barrier function in a murine atopic dermatitis-like dermatitis.

BACKGROUND: Selective activator protein (AP)-1 inhibitors are potentially promising therapeutic agents for atopic dermatitis (AD) because AP-1 is an important regulator of skin inflammation. However, few studies have investigated the effect of topical application of AP-1 inhibitors in treating inflammatory skin disorders. METHODS: Immunohistochemistry was conducted to detect phosphorylated AP-1/c-Jun expression of skin lesions in AD patients. In the in vivo study, 1 % T-5224 ointment was topically applied for 8 days to the ears of 2,4 dinitrofluorobenzene challenged AD-like dermatitis model mice. Baricitinib, a conventional therapeutic agent Janus kinase (JAK) inhibitor, was also topically applied. In the in vitro study, human epidermal keratinocytes were treated with T-5224 and stimulated with AD-related cytokines. RESULTS: AP-1/c-Jun was phosphorylated at skin lesions in AD patients. In vivo, topical T-5224 application inhibited ear swelling (P < 0.001), restored filaggrin (Flg) expression (P < 0.01), and generally suppressed immune-related pathways. T-5224 significantly suppressed Il17a and l17f expression, whereas baricitinib did not. Baricitinib suppressed Il4, Il19, Il33 and Ifnb expression, whereas T-5224 did not. Il1a, Il1b, Il23a, Ifna, S100a8, and S100a9 expression was cooperatively downregulated following the combined use of T-5224 and baricitinib. In vitro, T-5224 restored the expression of FLG and loricrin (LOR) (P < 0.05) and suppressed IL33 expression (P < 0.05) without affecting cell viability and cytotoxicity. CONCLUSIONS: Topical T-5224 ameliorates clinical manifestations of AD-like dermatitis in mice. The effect of this inhibitor is amplified via combined use with JAK inhibitors.

Surfactant therapy using a bronchofiberscope in respiratory distress syndrome.

BACKGROUND: Avoiding endotracheal intubation and using nasal continuous positive airway pressure as the initial treatment is recommended in infants with respiratory distress syndrome (RDS), and modes of lesser invasive surfactant administration have recently been reported. We report a pilot study assessing the feasibility of surfactant therapy using a bronchofiberscope (STUB) in RDS. METHODS: Surfactant was administered to 31 preterm infants (gestational age range of 28 weeks 0 days to 36 weeks 6 days) diagnosed with RDS, through the working channel of the bronchofiberscope or endotracheal tubes. Patient characteristics, outcomes, adverse events, and comorbidities were assessed in the two groups. RESULTS: Twelve infants received STUB. Two of the 12 infants (17%) needed subsequent intubation and additional surfactant administration. Nineteen infants received surfactant through endotracheal tubes. Four of the 19 infants (21%) required additional surfactants. There was no significant difference in the number of infants that needed additional surfactant (p = 1.00). Gestational age, birthweight, length of hospitalization, adverse events, such as desaturations and bradycardias, and comorbidities were similar between the two groups. Days of invasive ventilation were significantly shorter in the STUB group (p = 0.0002). CONCLUSION: STUB was feasible in this small cohort and reduced the need for intubation to 17%, leading to fewer days of invasive ventilation, without increasing comorbidities and adverse events. To the best of our knowledge, this is the first study to administer surfactants using bronchofiberscopes.

Ceramide Analysis in Combination With Genetic Testing May Provide a Precise Diagnosis for Self-Healing Collodion Babies.

Self-healing collodion baby (SHCB), also called "self-improving collodion baby", is a rare mild variant of autosomal recessive congenital ichthyosis and is defined as a collodion baby who shows the nearly complete resolution of scaling within the first 3 months to 1 year of life. However, during the neonatal period, it is not easy to distinguish SHCB from other inflammatory forms of autosomal recessive congenital ichthyosis, such as congenital ichthyosiform erythroderma. Here, we report a case study of two Japanese SHCB patients with compound heterozygous mutations, c.235G>T (p.(Glu79∗))/ c.1189C>T (p.(Arg397Cys)) and c.1295A>G (p.(Tyr432Cys))/ c.1138delG (p.(Asp380Thrfs∗3)), in CYP4F22, which encodes cytochrome P450, family 4, subfamily F, polypeptide 22 (CYP4F22). Immunohistochemically, inflammation with the strong expression of IL-17C, IL-36γ, and TNF-α was seen in the skin at birth. CYP4F22 is an ultra-long-chain FA ω-hydroxylase responsible for ω-O-acylceramide (acylceramide) production. Among the epidermal ceramides, acylceramide is a key lipid in maintaining the epidermal permeability barrier function. We found that the levels of ceramides with ω-hydroxy FAs including acylceramides and the levels of protein-bound ceramides were much lower in stratum corneum samples obtained by tape stripping from SHCB patients than in those from their unaffected parents and individuals without SHCB. Additionally, our cell-based enzyme assay revealed that two mutants, p.(Glu79∗) and p.(Arg397Cys), had no enzyme activity. Our findings suggest that genetic testing coupled with noninvasive ceramide analyses using tape-stripped stratum corneum samples might be useful for the early and precise diagnosis of congenital ichthyoses, including SHCB.

Obesity and Dyslipidemia Synergistically Exacerbate Psoriatic Skin Inflammation.

Patients with psoriasis are frequently complicated with metabolic syndrome; however, it is not fully understood how obesity and dyslipidemia contribute to the pathogenesis of psoriasis. To investigate the mechanisms by which obesity and dyslipidemia exacerbate psoriasis using murine models and neonatal human epidermal keratinocytes (NHEKs), we used wild-type and Apoe-deficient dyslipidemic mice, and administered a high-fat diet for 10 weeks to induce obesity. Imiquimod was applied to the ear for 5 days to induce psoriatic dermatitis. To examine the innate immune responses of NHEKs, we cultured and stimulated NHEKs using IL-17A, TNF-α, palmitic acid, and leptin. We found that obesity and dyslipidemia synergistically aggravated psoriatic dermatitis associated with increased gene expression of pro-inflammatory cytokines and chemokines. Treatment of NHEKs with palmitic acid and leptin amplified pro-inflammatory responses in combination with TNF-α and IL-17A. Additionally, pretreatment with palmitic acid and leptin enhanced IL-17A-mediated c-Jun N-terminal kinase phosphorylation. These results revealed that obesity and dyslipidemia synergistically exacerbate psoriatic skin inflammation, and that metabolic-disorder-associated inflammatory factors, palmitic acid, and leptin augment the activation of epidermal keratinocytes. Our results emphasize that management of concomitant metabolic disorders is essential for preventing disease exacerbation in patients with psoriasis.

Examination of changes in the breathing intolerance index to determine the respiratory support needs of very low birth weight infants.

BACKGROUND: The breathing intolerance index (BITI) is used to evaluate respiratory muscle tolerance. The higher the ratio of the inspiratory time to the total breathing time and the ratio of the tidal volume to the vital capacity, the more easily the respiratory muscles become fatigued. The BITI is high with chronic respiratory failure, and values of 0.15 or more indicate the need for assisted ventilation. OBJECTIVE: This study aimed to examine the changes in the BITI of very low birth weight infants (VLBWIs) and determine whether it is possible to use the BITI as an objective indicator of the timing of noninvasive respiratory support (NRS) withdrawal. METHODS: VLBWIs admitted to our neonatal intensive care unit between July 2020 and July 2022 under NRS at 36 weeks' postmenstrual age (PMA). If the BITI was less than 0.15, then we attempted to determine whether NRS could be withdrawn. RESULTS: Sixteen infants with a median gestational age and birth weight of 30.9 weeks and 1249 g, respectively, were eligible. The median PMA at the time of the first examination was 36.6 weeks. The BITI of two VLBWIs was less than 0.15. For 11 of the other VLBWIs, the BITI decreased over time to less than 0.15 at 39 weeks' PMA. After confirming that the BITI was less than 0.15, weaning from NRS was attempted. Weaning from NRS was possible except one VLBWI. CONCLUSION: The BITI can help evaluate respiratory tolerance and could be an objective indicator of the timing of NRS withdrawal.

Airway diseases in very low birth weight infants.

OBJECTIVE: To identify the incidence and characteristics of airway diseases in very low birth weight infants (VLBWIs). METHODS: A retrospective study of 214 inborn VLBWIs admitted to our NICU between April 2009 and March 2022 (approval no: 2023-0008). Neonatologists ourselves performed bronchoscopy to diagnose airway diseases. RESULTS: Symptomatic airway diseases were present in 36/214 (16.8%) of VLBWIs. Common airway diseases were tracheobronchomalacia (TBM) and pharyngomalacia. Infants with airway diseases had shorter gestational age, lower birth weight, more boys, and more moderate/severe bronchopulmonary dysplasia (BPD). Regarding treatment, more infants with airway diseases required intubation were intubated longer, used more dexamethasone, were on ventilators and oxygen longer, and were hospitalized longer. CONCLUSION: We found that VLBWIs were more frequently complicated with airway diseases, especially TBM. We also observed many pharyngeal lesions, which have not been previously reported. Intensity of prematurity, BPD, and the need for stronger respiratory management were risks for airway diseases. In VLBWIs, bronchoscopy should be actively performed because airway diseases are important complications.

Co-occurrence of non-alcoholic steatohepatitis exacerbates psoriasis associated with decreased adiponectin expression in a murine model.

Introduction: Clinical studies have suggested a bidirectional association between non-alcoholic steatohepatitis (NASH) and psoriasis, affecting each other's development and severity. Here, we explored bidirectional causal linkages between NASH and psoriasis using a murine model. Methods: NASH was induced in mice by streptozotocin injection at 2 days of age and by high-fat diet feeding (STAM™ model). Psoriasis was induced by topical application of imiquimod (IMQ) on the ear. The severities of liver damage and psoriatic skin changes were determined using histological analysis. Gene expression in the skin tissues was evaluated using quantitative PCR analysis. Serum cytokine levels were determined using enzyme-linked immunosorbent assay. To examine the innate immune responses of normal human epidermal keratinocytes (NHEKs), the cells were treated with interleukin (IL)-17A, tumor necrosis factor (TNF)-α, and AdipoRon, an adiponectin receptor agonist. Results and Discussion: There were no differences in the degree of liver tissue damage (fat deposition, inflammation, and fibrosis) between NASH mice with and those without psoriasis. Conversely, the co-occurrence of NASH significantly augmented psoriatic skin changes, represented by epidermal hyperplasia, in psoriatic mice. Pro-inflammatory cytokines were expressed in the inflamed skin of psoriatic mice, and the expression of genes, especially Il23a, Il1b, Il36g, and Mip2, was significantly upregulated by the co-occurrence of NASH. The expression of keratinocyte activation marker genes Defb4b and Krt16 was also upregulated by the co-occurrence of NASH. The serum TNF-α and IL-17 levels were increased by the co-occurrence of NASH and psoriasis. The serum adiponectin levels decreased in NASH mice compared with that in non-NASH mice. In NHEK culture, TNF-α and IL-17A synergistically upregulated CXCL1, CXCL8, and IL1B expression. The upregulated pro-inflammatory gene expression was suppressed by AdipoRon treatment, reflecting the anti-inflammatory capacity of adiponectin. Conclusion: The co-occurrence of NASH exacerbated psoriatic skin changes associated with increased serum inflammatory cytokine levels and decreased serum adiponectin levels. Combined with in vitro findings, increased inflammatory cytokine levels and decreased adiponectin levels likely promote innate immune responses in epidermal keratinocytes in psoriatic skin lesions. Overall, therapeutic intervention for co-occurring NASH is essential to achieve a favorable prognosis of psoriasis in clinical practice.

Survival rates of systemic interventions for psoriasis in the Western Japan Psoriasis Registry: A multicenter retrospective study.

Psoriasis affects approximately 0.3% of the Japanese population. Recently, various effective systemic drugs have become available, and the continuation of a given treatment has become critical because of the chronic nature of psoriasis. Factors affecting drug survival (the time until treatment discontinuation) in psoriasis treatment include efficacy, safety, ease of use, and patient preference. In the present study, the authors retrospectively surveyed a multifacility patient registry to determine the real-world evidence of the survival rate of systemic interventions for psoriasis treatment. Patients with psoriasis who visited 20 facilities in the Western Japan area between January 2019 and May 2020 and gave written consent were registered as study participants, and their medical history of systemic interventions for psoriasis (starting from 2010) was retrospectively collected and analyzed. The drugs investigated were adalimumab, infliximab, ustekinumab, secukinumab, ixekizumab, brodalumab, guselkumab, risankizumab, cyclosporine, and apremilast. When drugs were discontinued, the reasons were also recorded. A total of 1003 patients with psoriasis including 268 with psoriatic arthritis (PsA) were enrolled. In biologics, more recently released drugs such as interleukin 17 inhibitors showed a numerically higher survival rate in the overall (post-2010) analysis. However, in the subset of patients who began treatment after 2017, the difference in the survival rate among the drugs was smaller. The reasons for discontinuing drugs varied, but a loss of efficacy against dermatological or joint symptoms were relatively frequently seen with some biologics and cyclosporine. The stratification of drug survival rates based on patient characteristics such as bio-naive or experienced, normal weight or obese, and with or without PsA, revealed that bio-experienced, obese, and PsA groups had poorer survival rates for most drugs. No notable safety issues were identified in this study. Overall, the present study revealed that the biologics show differences in their tendency to develop a loss of efficacy, and the factors that negatively impact the survival rate of biologics include the previous use of biologics, obesity, and PsA.

TRAPS mutations in Tnfrsf1a decrease the responsiveness to TNFα via reduced cell surface expression of TNFR1.

Tumor necrosis factor (TNF) receptor-associated periodic syndrome (TRAPS) is an autoinflammatory periodic fever syndrome associated with heterozygous mutations in TNFRSF1A, which encodes TNF receptor type I (TNFR1). Although possible proinflammatory mechanisms have been proposed, most previous studies were performed using in vitro overexpression models, which could lead to undesirable inflammatory responses due to artificial overexpression. It is crucial to reproduce heterozygous mutations at physiological expression levels; however, such studies remain limited. In this study, we generated TRAPS mutant mice and analyzed their phenotypes. Three Tnfrsf1a mutant strains were generated by introducing T79M, G87V, or T90I mutation. T79M is a known mutation responsible for TRAPS, whereas G87V is a TRAPS mutation that we have reported, and T90I is a variant of unknown significance. Using these murine models, we investigated whether TRAPS mutations could affect the inflammatory responses in vivo and in vitro. We found that none of the mutant mice exhibited detectable inflammatory phenotypes under standard housing conditions for 1 year. Interestingly, TRAPS mutant (T79M and G87V) mice had reduced mortality rates after the administration of lipopolysaccharide (LPS) and D-galactosamine, which induce TNFα-dependent lethal hepatitis. Moreover, TRAPS mutations strongly suppressed the development of TNFα-mediated arthritis when crossed with human TNFα transgenic mice. In in vitro primary bone marrow-derived macrophage cultures, the T79M and G87V mutations attenuated the inflammatory responses to TNFα compared with the wild-type, whereas these mutations did not alter the responsiveness of these cells to LPS. The T90I mutant macrophages behaved similarly to wild type in response to LPS and TNFα. The TNFR1 levels were increased in whole-cell lysates of TRAPS mutant macrophages, whereas the cell surface expression of TNFR1 was significantly decreased in TRAPS mutant macrophages. Taken together, TRAPS mutations did not augment the inflammatory responses to TNFα and LPS; instead, they suppressed the response to TNFα via decreased cell surface expression of TNFR1. The stimulation of lymphotoxin-α, adenosine triphosphate, and norepinephrine in primary macrophages or various stimuli in murine splenocytes did not induce detectable inflammatory responses. In conclusion, TRAPS mutations suppressed responsiveness to TNFα, and TRAPS-associated inflammation is likely induced by unconfirmed disease-specific proinflammatory factors.

Campylobacter fetus Meningitis and Pyogenic Spondylodiscitis in a Healthy Young Woman.

We report a rare case of Campylobacter fetus meningitis and pyogenic spondylodiscitis in a healthy young woman. A 35-year-old woman without significant medical history presented with fever, headache, and low back pain. C. fetus was detected from the blood culture. Cerebrospinal fluid analysis showed bacterial meningitis 2 days after onset. Although initial magnetic resonance imaging (MRI) did not reveal abnormal findings, repeated MRI showed a low-signal-intensity lesion on T1-weighted image (T1WI) and a high-signal-intensity lesion on T2WI between the L5 and S1 vertebral bodies 11 days after onset. The mode of infection was considered to be the consumption of raw chicken meat. After antibiotic treatment with 12 g/day ampicillin following 6 g/day meropenem, she was discharged 51 days after onset. As the inflammatory signs on MRI improved, oral antibiotic therapy was discontinued 85 days after onset. Although the initial MRI showed no abnormal findings, repeat MRI should be performed for patients who have persistent low back pain and fever.

Syntheses of four-membered metallacyclic complexes with nitrosylruthenium and their ring-opening upon HCl addition.

Symmetrically disubstituted bis(3-hydroxyalkynyl) complex [TpRu{C[triple chemical bond]CCPh(2)(OH)}(2)(NO)] (1) (Tp = BH(pyrazol-1-yl)(3)) and unsymmetrically mixed (arylalkynyl)(3-hydroxyalkynyl) congener [TpRu(C[triple chemical bond]CC(6)H(4)Me){C[triple chemical bond]CCPh(2)(OH)}(NO)] (2) were newly prepared. Treatment of 1 or 2 with p-toluenesulfonic acid monohydrate was carried out to give unusual four-membered metallacyclic complexes [TpRu{C(=C=CPh(2))C(O)C(=CPh(2))}(NO)] (3) and [TpRu{C(=C=CPh(2))C(O)CH(C(6)H(4)Me)}(NO)] (5), respectively, as major products. Formation mechanism of 3 and 5 would involve insertion of the generated allenylidene group (Ru=C=C=CPh(2)) into the other Ru--C(alkynyl) bond, followed by hydration of the resulting alpha-alkynyl--allenyl fragment. With regards to the chemical reactivity of their four-membered metallacycles, treatment with aq. HCl in MeOH afforded the ring-opened one-HCl adducts, [TpRuCl{C(=C=CPh(2))C(O)CH=CPh(2)}(NO)] (7) and [TpRuCl{C(=C=CPh(2))C(O)CH(2)(C(6)H(4)Me)}(NO)] (8). On the other hand, the use of CH(2)Cl(2) and THF as the reaction solvent gave another type of one-HCl adducts [TpRu{CH(C(Cl)=CPh(2))C(O)C(==CPh(2))}(NO)] (9 a/9 b) and [TpRu{CH(C(Cl)=CPh(2))C(O)CH(C(6)H(4)Me)}(NO)] (11 a/11 b) as diastereomeric pairs, still retaining the four-membered ring structure. Moreover, their kinetically controlled products 9 b and 11 b were treated with aq. HCl to afford the ring-opened two-HCl adducts [TpRuCl{C(C(Cl)=CPh(2))(H)C(O)CH=CPh(2)}(NO)] (10) and [TpRuCl{CH(C(6)H(4)Me)C(O)CH(2)(C(Cl)=CPh(2))}(NO)] (12), respectively. In 10 and 12, each one Ru--C bond is cleaved at mutually different positions in the ring. Protonation on the carbonyl group would trigger the formation of 7-12.

Hydration of nitrosylruthenium bis(alkynyl) complexes with hydrotris(pyrazolyl)borate: insertion/hydration and double hydration products.

Hydration of nitrosylruthenium bis(alkynyl) complex TpRu(CCPh)2(NO) (1) (Tp = BH(pyrazol-1-yl)3) was carried out in the presence of HBF4.Et2O in distilled MeOH and afforded the metallacycle TpRu{CH=C(Ph)C(O)CH(Ph)}(NO) (2) (39%) and the bis(ketonyl) TpRu(CH2C(O)Ph)2(NO) (3) (37%). While double hydration of 1 gave 3, 2 was produced through a combination of insertion and hydration processes. On the other hand, a similar reaction performed in THF instead of MeOH afforded 2 (52%), the acyl-ketonyl complex TpRu(C(O)CH2Ph)(CH2C(O)Ph)(NO) (4) (8.9%), and trace amounts of 3 and TpRu(CCPh)(CH2C(O)Ph)(NO) (5). Moreover, the 1/HBF4.Et2O/H2O reaction system in distilled MeOH at 0 degrees C gave rise to 5 exclusively (79%). Treatment of THF solution of isolated 5 with water in the presence of protic acid furnished 3 and 4, revealing that 5 is the intermediate in their formation.