Bullous Pemphigoid IgG Induces BP180 Internalization via a Macropinocytic Pathway

Bullous pemphigoid (BP) is an autoimmune blistering skin disease induced by pathogenic autoantibodies against a type II transmembrane protein (BP180, collagen type XVII, or BPAG2). In animal models, BP180 autoantibody-antigen interaction appears insufficient to develop blisters, but involvement of complement and neutrophils is required. However, cultured keratinocytes treated with BP-IgG exhibit a reduction in the adhesive strength and a loss of expression of BP180, suggesting that the autoantibodies directly affect epidermal cell–extracellular matrix integrity. In this study, we explored the consequences of two distinct epithelial cells treated with BP-IgG, particularly the fate of BP180. First, we followed the distribution of green fluorescent protein–tagged BP180 in an epithelial cell line, 804G, and normal human epidermal keratinocytes after autoantibody clustering. After BP-IgG treatment, the adhesive strength of the cells to their substrate was decreased, and BP180 was internalized in both cell types, together with the early endosomal antigen-1. By using various endocytosis inhibitors and a fluid-uptake assay, we demonstrated that BP-IgG–induced BP180 internalization is mediated via a macropinocytic pathway. Moreover, a macropinocytosis inhibitor rescued a BP-IgG–induced reduction in the adhesive strength of the cells from their substrate. The results of this study suggest that BP180 internalization induced by BP-IgG plays an important role in the initiation of disease pathogenesis

Prenatal Vitamin E Treatment Improves Lung Growth in Fetal Rats with Congenital Diaphragmatic Hernia

The aim of this study was to test the effects of vitamin E on fetal lung growth in rats with congenital diaphragmatic hernia (CDH). Experimental congenital diaphragmatic hernia (CDH) was induced in rat fetuses by maternal administration of 100 mg nitrofen by gastric gavage on day 9.5 of gestation. Vitamin E was provided at days 16-20 of gestation, at 30 IU/day. Cesarean section was performed at day 21 of gestation. Immunohistochemistry was performed using anti-surfactant protein A (SP-A) and anti-SP-B polyclonal antibodies. RT-PCR evaluated SP-A and SP-B mRNA expressions. The lung weight/body weight ratio in rats with CDH was lower than the control (p<0.01). The number of type II pneumocytes positive for SP-A in untreated CDH rats (n=20) was lower than the control (n=20). The relative amounts of SP-A and SP-B were significantly higher in vitamin Etreated CDH rats (n=20) than untreated CDH rats (p<0.05). Our results suggest that antenatal vitamin E treatment increases the production of surfactant proteins in hypoplastic lung of rats with the CDH

Simultaneous Three-Dimensional Analysis of Cervical Spine Kinematics in the Axial and Sagittal Views during a Simulated Frontal Impact: Differences between Tensed and Relaxed States

Study DesignProspective experimental study on humans.PurposeTo determine whether postural differences during a low-speed impact are observed in the sagittal and axial views, particularly in a relaxed state.Overview of Literature: Three-dimensional motion capture systems have been used to analyze posture and head-neck-torso kinematics in humans during a simulated low-speed impact, yet little research has focused on the axial view. Since a seatbelt asymmetrically stabilizes a drivers right shoulder and left lower waist into the seat, it potentially creates movement in the axial view.MethodsThree healthy adult men participated in the experimental series, which used a low-speed sled system. The acceleration pulse created a full sine shape with a maximum acceleration of 8.0 m/s2 at 500 ms, during which the kinematics were evaluated in relaxed and tensed states. The three-dimensional motion capture system used eight markers to record and analyze body movement and head-neck-torso kinematics in the sagittal and axial views during the low-speed impact. Head and trunk rotation angles were also calculated.ResultsLarger movements were observed in the relaxed than in the tensed state in the sagittal view. The cervical and thoracic spine flexed and extended, respectively, in the relaxed state. In the axial view, larger movements were also observed in the relaxed state than in the tensed state, and the left shoulder rotated.ConclusionsDuring simulated frontal impact, the rotation angle between the head and trunk was significantly larger in the relaxed state. Therefore, we recommend also observing movement in the axial view during impact tests

Preoperative diagnosis of pelvic actinomycosis by clinical cytology

Background: The purpose of this work was to investigate whether clinical cytology could be useful in the preoperative diagnosis of pelvic actinomycosis. Methods: This study involved the prospective collection of samples derived from the endometrium and the uterine cervix, and retrospective data analysis. Nine patients with clinically diagnosed pelvic actinomycosis were enrolled. The clinical and hematological characteristics of patients were recorded, and detection of actinomyces was performed by cytology, pathology, and bacteriological culture of samples and by imprint intrauterine contraceptive device (IUD) cytology. Results: The detection rate of actinomyces was 77.7% by combined cervical and endometrial cytology, 50.0% by pathology, and 11.1% by bacterial culture. Conclusion: The higher detection rate of actinomyces by cytology than by pathology or bacteriology suggests that careful cytological examination may be clinically useful in the preoperative diagnosis of pelvic actinomycosis

Imbalanced expression of polycistronic miRNA in acute myeloid leukemia

miR-1 and miR-133 are clustered on the same chromosomal loci and are transcribed together as a single transcript that is positively regulated by ecotropic virus integration site-1 (EVI1). Previously, we described how miR-133 has anti-tumorigenic potential through repression of EVI1 expression. It has also been reported that miR-1 is oncogenic in the case of acute myeloid leukemia (AML). Here, we show that expression of miR-1 and miR-133, which have distinct functions, is differentially regulated between AML cell lines. Interestingly, the expression of miR-1 and EVI1, which binds to the promoter of the miR-1/miR-133 cluster, is correlative. The expression levels of TDP-43, an RNA-binding protein that has been reported to increase the expression, but inhibits the activity, of miR-1, were not correlated with expression levels of miR-1 in AML cells. Taken together, our observations raise the possibility that the balance of polycistronic miRNAs is regulated post-transcriptionally in a hierarchical manner possibly involving EVI1,suggesting that the deregulation of this balance may play some role in AML cells with high EVI1 expression

Functional roles of Aves class-specific cis-regulatory elements on macroevolution of bird-specific features

Unlike microevolutionary processes, little is known about the genetic basis of macroevolutionary processes. One of these magnificent examples is the transition from non-avian dinosaurs to birds that has created numerous evolutionary innovations such as self-powered flight and its associated wings with flight feathers. By analysing 48 bird genomes, we identified millions of avian-specific highly conserved elements (ASHCEs) that predominantly (>99%) reside in non-coding regions. Many ASHCEs show differential histone modifications that may participate in regulation of limb development. Comparative embryonic gene expression analyses across tetrapod species suggest ASHCE-associated genes have unique roles in developing avian limbs. In particular, we demonstrate how the ASHCE driven avian-specific expression of gene Sim1 driven by ASHCE may be associated with the evolution and development of flight feathers. Together, these findings demonstrate regulatory roles of ASHCEs in the creation of avian-specific traits, and further highlight the importance of cis-regulatory rewiring during macroevolutionary changes

Poster Session

International Symposium on Tumor Biology in Kanazawa & Symposium on Drug Discoverry in Academics 2014 [DATE]: January 23(Thu)-24(Fri),2014, [Place]:Kanazawa Excel Hotel Tpkyu, Kanazawa, Japan, [Organizers]:Kanazawa Association of Tumor Biologists / Cancer Research Institute, Kanazawa Universit

Minimizing Errors in RT-PCR Detection and Quantification of SARS-CoV-2 RNA for Wastewater Surveillance

Wastewater surveillance for pathogens using the reverse transcription-polymerase chain reaction (RT-PCR) is an effective, resource-efficient tool for gathering additional community-level public health information, including the incidence and/or prevalence and trends of coronavirus disease-19 (COVID-19). Surveillance of SARS-CoV-2 in wastewater may provide an early-warning signal of COVID-19 infections in a community. The capacity of the world’s environmental microbiology and virology laboratories for SARS-CoV-2 RNA characterization in wastewater is rapidly increasing. However, there are no standardized protocols nor harmonized quality assurance and quality control (QA/QC) procedures for SARS-CoV-2 wastewater surveillance. This paper is a technical review of factors that can lead to false-positive and -negative errors in the surveillance of SARS-CoV-2, culminating in recommendations and strategies that can be implemented to identify and mitigate these errors. Recommendations include, stringent QA/QC measures, representative sampling approaches, effective virus concentration and efficient RNA extraction, amplification inhibition assessment, inclusion of sample processing controls, and considerations for RT-PCR assay selection and data interpretation. Clear data interpretation guidelines (e.g., determination of positive and negative samples) are critical, particularly during a low incidence of SARS-CoV-2 in wastewater. Corrective and confirmatory actions must be in place for inconclusive and/or potentially significant results (e.g., initial onset or reemergence of COVID-19 in a community). It will also be prudent to perform inter-laboratory comparisons to ensure results are reliable and interpretable for ongoing and retrospective analyses. The strategies that are recommended in this review aim to improve SARS-CoV-2 characterization for wastewater surveillance applications. A silver lining of the COVID-19 pandemic is that the efficacy of wastewater surveillance was demonstrated during this global crisis. In the future, wastewater will play an important role in the surveillance of a range of other communicable diseases.Highlights: Harmonized QA/QC procedures for SARS-CoV-2 wastewater surveillance are lacking; Wastewater analysis protocols are not optimized for trace analysis of viruses; False-positive and -negative errors have consequences for public health responses; Inter-laboratory studies utilizing standardized reference materials and protocols are needed.info:eu-repo/semantics/publishedVersio