Simultaneous Disruption of Mouse ASIC1a, ASIC2 and ASIC3 Genes Enhances Cutaneous Mechanosensitivity

Three observations have suggested that acid-sensing ion channels (ASICs) might be mammalian cutaneous mechanoreceptors; they are structurally related to Caenorhabditis elegans mechanoreceptors, they are localized in specialized cutaneous mechanosensory structures, and mechanical displacement generates an ASIC-dependent depolarization in some neurons. However, previous studies of mice bearing a single disrupted ASIC gene showed only subtle or no alterations in cutaneous mechanosensitivity. Because functional redundancy of ASIC subunits might explain limited phenotypic alterations, we hypothesized that disrupting multiple ASIC genes would markedly impair cutaneous mechanosensation. We found the opposite. In behavioral studies, mice with simultaneous disruptions of ASIC1a, -2 and -3 genes (triple-knockouts, TKOs) showed increased paw withdrawal frequencies when mechanically stimulated with von Frey filaments. Moreover, in single-fiber nerve recordings of cutaneous afferents, mechanical stimulation generated enhanced activity in A-mechanonociceptors of ASIC TKOs compared to wild-type mice. Responses of all other fiber types did not differ between the two genotypes. These data indicate that ASIC subunits influence cutaneous mechanosensitivity. However, it is unlikely that ASICs directly transduce mechanical stimuli. We speculate that physical and/or functional association of ASICs with other components of the mechanosensory transduction apparatus contributes to normal cutaneous mechanosensation

Simultaneous Detection of EGFR and VEGF in Colorectal Cancer using Fluorescence-Raman Endoscopy

Fluorescence endomicroscopy provides quick access to molecular targets, while Raman spectroscopy allows the detection of multiple molecular targets. Using a simultaneous fluorescence-Raman endoscopic system (FRES), we herein demonstrate its potential in cancer diagnosis in an orthotopically induced colorectal cancer (CRC) xenograft model. In the model, epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) were targeted with antibody-conjugated fluorescence and surface-enhanced Raman scattering (F-SERS) dots. FRES demonstrated fast signal detection and multiplex targeting ability using fluorescence and Raman signals to detect the F-SERS dots. In addition, FRES showed a multiplex targeting ability even on a subcentimeter-sized CRC after spraying with a dose of 50 µg F-SERS dots. In conclusion, molecular characteristics of tumor cells (EGFR in cancer cell membranes) and tumor microenvironments (VEGF in the extracellular matrix) could be simultaneously investigated when performing a colonoscopy

H₂O₂-induced nociceptive behavior in rats as total time spent flinching, lifting and licking of the hind leg.

<p>(A) Spontaneous nociceptive behavior after intramuscular (n = 6) or subcutaneous (n = 5) injection of H₂O₂ (100 mM, 0.6 ml), or intramuscular injection of synthetic interstitial fluid (0.6 ml) (n = 6). * P = 0.0004 compared with control group, † P = 0.0010 compared with subcutaneous H₂O₂ injection group by one-way ANOVA followed by post-hoc Bonferroni’s test. (B) Effects of local pre-injection of a TRPA1 antagonist (HC-030031; 50 mM, 0.3 ml, n = 6) on nociceptive behavior caused by intramuscular injection of H₂O₂ (100 mM, 0.3 ml, n = 7) vs. vehicle group (n = 7). * P < 0.0001 compared with vehicle + PBS injection group, † P < 0.0001 compared with HC-030031 + H₂O₂ injection group by one-way ANOVA followed by post-hoc Bonferroni’s test. All data are expressed as means ± SEM.</p

Effect of intraperitoneal (i.p.) administration of HC-030031 on pain behaviors of rats after skin + deep tissue incision.

<p>(A) Guarding pain behavior. The results are presented as mean and standard error of the mean (SEM) for eight rats in each group. Two-way ANOVA with repeated measures on one factor (interaction factor: F_{24, 210} = 1.64, P = 0.0360) followed by Bonferroni’s post hoc test for comparing the mean cumulative pain score at each time point among groups. (B) Withdrawal threshold to punctate stimuli applied to the hind paw. The results are presented as median with range for six rats in each group. Non-parametric Friedman’s test (Fr = 17.02, P = 0.0019) followed by Kruskal-Wallis test with Dunn post hoc test for between-group comparisons at each time point. (C) Withdrawal latency to heat stimulation. The results are presented as mean and SEM. Two-way ANOVA with repeated measures on one factor (interaction factor: F_{24, 150} = 1.163, P = 0.2855) followed by Bonferroni’s post hoc test for comparing the mean withdrawal latency at each time point among groups. * P < 0.05, ** P < 0.01, † P < 0.001 compared with the vehicle group at each time point. POD = postoperative day.</p

Effects of the incision of skin and muscle on tissue hydrogen peroxide (H₂O₂) levels in rats.

<p>(A) H₂O₂ content after incision in gastrocnemius muscle using the Amplex^® Red Hydrogen Peroxide assay kit. The results are presented as mean and SEM for 6 rats in each group. Two-way ANOVA with repeated measures on one factor (interaction factor: F_{2, 15} = 2.328, P = 0.1317, Time factor: F_{2, 15} = 3.890, P = 0.0436, Group factor: F_{1, 15} = 22.58, P = 0.0003) followed by Bonferroni’s post hoc test. * P < 0.0001 compared with non-incised muscle on POD 0, † P = 0.0074 compared with non-incised muscle on POD 1. (B) H₂O₂ content after incision of skin overlying the gastrocnemius muscle. The results are presented as mean and SEM for 6 rats in each group. Two-way ANOVA with repeated measures on one factor (interaction factor: F_{2, 15} = 0.5907, P = 0.5663, Time factor: F_{2, 15} = 0.2134, P = 0.8103, Group factor: F_{1, 15} = 31.71, P < 0.0001) followed by Bonferroni’s post hoc test. # P = 0.0105 compared with incised skin on POD 0, ‡ P = 0.0373 compared with incised skin on POD 1.</p

<i>In vivo</i> reactive oxygen species (ROS)-imaging with L-012 after incision in rats.

<p>(A) Examples of <i>in vivo</i> imaging after gastrocnemius muscle incision. (B) Average luminescence intensity in <i>in vivo</i> ROS-imaging on gastrocnemius muscle incision, gastrocnemius incision with catalase (1,000–2,500 IU), skin-only incision, and sham. The results are presented as mean and SEM for 6 rats in each group. * P < 0.0001 compared with sham, † P < 0.0001 compared with gastrocnemius incision with catalase, ‡ P < 0.0001 compared with skin-only incision, # P < 0.0001 compared with sham, § P = 0.0159 compared with gastrocnemius incision with catalase, & P = 0.0490 compared with skin-only incision, ¶ P = 0.0166 compared with sham. Two-way ANOVA (interaction factor: F_{6, 30} = 22.56, P < 0.0001) followed by Bonferroni's post hoc tests.</p

Ongoing increasing trends in central precocious puberty incidence among Korean boys and girls from 2008 to 2020.

BackgroundOver the last few decades, there has been growing evidence of earlier onset and progression of puberty worldwide. This population-based longitudinal cohort study aimed to analyze the change in the annual incidence rate of central precocious puberty (CPP) among Korean children over the most recent decade, using the national registry data.MethodThe International Statistical Classification of Diseases and Related Health Problems, Tenth Revision (ICD-10) and insurance claims for gonadotropin-releasing hormone agonist (GnRHa) treatment were used to identify CPP patients who were using the Korean Health Insurance Review & Assessment Service (HIRA) database between 2008 and 2020. Patients who began GnRHa therapy before the age of 9 and 10 for girls and boys, respectively, were included in the study.ResultsA total of 6,906 boys and 126,377 girls were diagnosed with CPP between 2008 and 2020. The annual incidence of CPP increased by 83.3 times in boys (from 1.2 to 100 per 100,000 persons) and by 15.9 times in girls (from 88.9 to 1414.7 per 100,000 persons). The age-specific annual incidence of CPP increased remarkably more in older children than in younger ones; the 2020 CPP incidence among 9-year-old boys and 8-year-old girls reached 705.2 and 7,967.3 per 100,000 persons, respectively. The annual prevalence of CPP in boys and girls increased from 2.7 to 206.5 (76.5 times) and from 141.8 to 3439.9 (24.3 times) per 100,000 persons, respectively.ConclusionBased on GnRHa treatment insurance claims, our study suggests that the annual incidence of CPP has substantially increased in Korea during the past 13 years. These findings highlight the importance of meticulous judgment by doctors in determining GnRHa treatment