Chemical Visualization of an Attractant Peptide, LURE

The pollen tube attractant peptide LUREs of Torenia fournieri are diffusible peptides that attract pollen tubes in vitro. Here, we report a method enabling the direct visualization of a LURE peptide without inhibiting its attraction activity by conjugating it with the Alexa Fluor 488 fluorescent dye. After purifying and refolding the recombinant LURE2 with a polyhistidine tag, its amino groups were targeted for conjugation with the Alexa Fluor dye. Labeling of LURE2 was confirmed by its fluorescence and mass spectrometry. In our in vitro assay using gelatin beads, Alexa Fluor 488-labeled LURE2 appeared to have the same activity as unlabeled LURE2. Using the labeled LURE2, the relationship between the spatiotemporal change of distribution and activity of LURE2 was examined. LURE2 attracted pollen tubes when embedded in gelatin beads, but hardly at all when in agarose beads. Direct visualization suggested that the significant difference between these conditions was the retention of LURE2 in the gelatin bead, which might delay diffusion of LURE2 from the bead. Direct visualization of LURE peptide may open the way to studying the spatiotemporal dynamics of LURE in pollen tube attraction

Comparison between Two Assessment Tests for Oral Hygiene : Adenosine Triphosphate + Adenosine Monophosphate Swab Test and Bacteria Number Counting by Dielectrophoretic Impedance Measurement

Objective assessments of oral hygiene are important to prevent oral and systemic diseases. Two objective assessment tests are available to assess oral hygiene; (1) the adenosine triphosphate (ATP) + adenosine monophosphate (AMP) swab test, which incorporates a luciferase assay and (2) a bacteria count using the dielectrophoretic impedance measurement (DEPIM) method. In this study, we compared the two tests using a subjective visual assessment by professional clinicians and investigated the clinical significance of these tests. Twenty-seven young participants (mean age 26.3 ± 3.2 years) and twenty-seven older participants (mean age 75.1 ± 5.9 years) were recruited. Oral bacteria were sampled from three areas, including the tongue dorsum, the buccal mucosa, and the faucal mucosa, and saliva was obtained using a cotton swab. The amount of ATP + AMP and the number of bacteria were measured by each specific apparatus. Additionally, one examiner assessed the overall condition of oral hygiene using the visual analog scale (VAS). In the ATP + AMP swab test, the means were highest in saliva. For the bacteria count, the means were higher in the tongue dorsum and saliva and lower in the faucal and buccal mucosa. The results of the subjective assessment of oral hygiene indicated that the VAS-value was 3.78 ± 0.97 for the young group and 3.35 ± 0.81 for the older group. No significant difference was observed between the two groups. Additionally, no significant relationship between the values of the ATP + AMP swab test and the bacteria count was found for any of the four sample sites. In the older group, the subjective assessment of oral hygiene was significantly correlated with the values of the ATP + AMP swab test (multiple correlation coefficient = 0.723, p = 0.002). In conclusion, the values provided by the ATP + AMP swab test were not always correlated to the bacteria count. The results of this study suggest that the subjective assessment of oral hygiene was more highly correlated with the results of the ATP + AMP swab test, as compared to the bacterial count assay

内反小趾変形のX線学的形態の特徴

Background: Bunionette deformity is characterized as head hypertrophy, lateral bowing, or splaying of the fifth metatarsal, or a combination of these deformities. Most previous studies have focused on the fourth and fifth metatarsals; few have analyzed the radiographic morphologic characteristics of the entire foot. The morphologic characteristics of the entire foot in cases of symptomatic bunionette deformity were analyzed with a radiographic image-mapping system. Methods: The system was used for the morphologic analysis of 112 feet with symptomatic deformity and 123 asymptomatic control feet. The mapping system includes 2-dimensional coordinates. We compared morphologies of both groups on the basis of simple models prepared from x and y coordinates of each reference point, calculated by using the mapping system and various angle measurements. We set cutoff values and categorized cases according to Fallat’s system. We evaluated the characteristics of each type and a new deformity type (type V) wherein no measurement exceeded the cutoff values. Results: The heads of the third, fourth, and fifth metatarsals were more laterally displaced, and the angles between the metatarsal axes were larger in the deformity group. Comparison of deformity types showed that the morphology of the fifth metatarsal might be only one cause of deformity. The intermetatarsal angles between the second and third metatarsals and between the third and fourth metatarsals were larger in deformity type II and type V feet than in control feet. Additionally, the intermetatarsal angles between the third and fourth metatarsals and between the fourth and fifth metatarsals were larger in deformity type III and type IV feet than in control feet. Conclusion: We believe it is necessary to not only focus on the fourth and fifth metatarsals, but also assess the morphologic characteristics of the entire foot in patients with a bunionette, including splaying of all the metatarsals and the forefoot width, when planning surgery. Level of Evidence: Level III, retrospective comparative study.博士（医学）・甲第651号・平成28年3月15日© The Author(s) 2015Copyright © 2016 by American Orthopaedic Foot & Ankle SocietyThe definitive version is available at " http://dx.doi.org/10.1177/1071100715614923

DOCK2 is involved in the host genetics and biology of severe COVID-19

「コロナ制圧タスクフォース」COVID-19疾患感受性遺伝子DOCK2の重症化機序を解明 --アジア最大のバイオレポジトリーでCOVID-19の治療標的を発見--. 京都大学プレスリリース. 2022-08-10.Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge. Here we conducted a genome-wide association study (GWAS) involving 2, 393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3, 289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2 associated with the risk allele in these younger patients. DOCK2 expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n = 61 individuals) identified cell-type-specific downregulation of DOCK2 and a COVID-19-specific decreasing effect of the risk allele on DOCK2 expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target

Comparison between Two Assessment Tests for Oral Hygiene: Adenosine Triphosphate + Adenosine Monophosphate Swab Test and Bacteria Number Counting by Dielectrophoretic Impedance Measurement

Objective assessments of oral hygiene are important to prevent oral and systemic diseases. Two objective assessment tests are available to assess oral hygiene; (1) the adenosine triphosphate (ATP) + adenosine monophosphate (AMP) swab test, which incorporates a luciferase assay and (2) a bacteria count using the dielectrophoretic impedance measurement (DEPIM) method. In this study, we compared the two tests using a subjective visual assessment by professional clinicians and investigated the clinical significance of these tests. Twenty-seven young participants (mean age 26.3 ± 3.2 years) and twenty-seven older participants (mean age 75.1 ± 5.9 years) were recruited. Oral bacteria were sampled from three areas, including the tongue dorsum, the buccal mucosa, and the faucal mucosa, and saliva was obtained using a cotton swab. The amount of ATP + AMP and the number of bacteria were measured by each specific apparatus. Additionally, one examiner assessed the overall condition of oral hygiene using the visual analog scale (VAS). In the ATP + AMP swab test, the means were highest in saliva. For the bacteria count, the means were higher in the tongue dorsum and saliva and lower in the faucal and buccal mucosa. The results of the subjective assessment of oral hygiene indicated that the VAS-value was 3.78 ± 0.97 for the young group and 3.35 ± 0.81 for the older group. No significant difference was observed between the two groups. Additionally, no significant relationship between the values of the ATP + AMP swab test and the bacteria count was found for any of the four sample sites. In the older group, the subjective assessment of oral hygiene was significantly correlated with the values of the ATP + AMP swab test (multiple correlation coefficient = 0.723, p = 0.002). In conclusion, the values provided by the ATP + AMP swab test were not always correlated to the bacteria count. The results of this study suggest that the subjective assessment of oral hygiene was more highly correlated with the results of the ATP + AMP swab test, as compared to the bacterial count assay