Utility of photodynamic diagnosis plus reflectance confocal microscopy in detecting the margins of extramammary Paget disease

Background: Due to the clinically poorly delineated unclear margin of extramammary Paget disease, the recurrence rate after surgical resection is high. Aims: To compare photodynamic diagnosis and photodynamic plus reflectance confocal microscopy diagnosis in determining the tumor margins in patients with extramammary Paget disease. Methods: Thirty-six patients with histopathologically confirmed primary extramammary Paget disease between January 2017 to June 2018 were included in the study. The skin lesion margins were preoperatively observed by the naked eye and with photodynamic diagnosis and photodynamic diagnosis plus reflectance confocal microscopy and they were compared to the postoperative histopathological examination results. Results: Among the 130 sections taken from 36 patients, 83 sections (63.8%, 83/130) had tumor margins beyond the macroscopic line with a distance of 3.5 ± 3.1mm and a median of 2.7mm. Forty-six sections (35.4%, 46/130) exceeded the photodynamic diagnosis marker line with a distance of 2.1 ± 1.7mm and a median of 1.5mm. Twenty seven sections (20.8%, 27/130) were obtained beyond the photodynamic diagnosis plus reflectance confocal microscopy marker line with a distance of 1.4 ± 1.2mm and a median of 0.9mm. Limitations: Photodynamic diagnosis and reflectance confocal microscopy detection can be used to observe only the superficial margin of the tumor and not the deep part. Moreover, reflectance confocal microscopy was not used alone as a control. Conclusion: In terms of determining the extramammary Paget disease margin invasively, photodynamic diagnosis and photodynamic diagnosis plus reflectance confocal microscopy were found superior to observations made with the naked eye, while photodynamic diagnosis plus reflectance confocal microscopy was superior to photodynamic diagnosis alone.

Dynamics of plant DNA replication based on PCNA visualization

DNA replication is an essential process for the copying of genomic information in living organisms. Imaging of DNA replication in tissues and organs is mainly performed using fixed cells after incorporation of thymidine analogs. To establish a useful marker line to measure the duration of DNA replication and analyze the dynamics of DNA replication, we focused on the proliferating cell nuclear antigen (PCNA), which functions as a DNA sliding clamp for replicative DNA polymerases and is an essential component of replisomes. In this study we produced an Arabidopsis thaliana line expressing PCNA1 fused with the green fluorescent protein under the control of its own promoter (pAtPCNA1::AtPCNA1-sGFP). The duration of the S phase measured using the expression line was consistent with that measured after incorporation of a thymidine analog. Live cell imaging revealed that three distinct nuclear localization patterns (whole, dotted, and speckled) were sequentially observable. These whole, dotted, and speckled patterns of subnuclear AtPCNA1 signals were indicative of the G1 or G2 phase, early S phase and late S phase, respectively. The results indicate that the pAtPCNA1::AtPCNA1-sGFP line is a useful marker line for visualization of S-phase progression in live plant organs.

New Radiographic Analysis of Sesamoid Rotation in Hallux Valgus: Comparison with Conventional Evaluation Methods

The position of the hallucal sesamoids needs to be included in evaluation of hallux valgus. In order to quantify the rotational position of the hallucal sesamoids, a new weightbearing tangential radiograph was established by means of a specially designed tangential positioning device. This device has a depression, and a tangential radiograph is taken with the metatarsophalangeal joint at 45° dorsiflexion. A lead marker plate is placed on the depression to show the horizontal plane, and the sesamoid rotation angle (SRA) is measured. The SRA is the angle between the tangential line of the most inferior aspect of the medial-lateral sesamoids and the lead marker line. The SRA was compared with values of the four-grade scale and seven-position scale which were measured from the antero-posterior view, with respect to the hallux valgus angle (HVA), by means of conventional methods. Measurements were made of 58 feet in 29 patients with hallux valgus and 64 feet in 32 normal subjects. The SRA showed the highest correlation among the three parameters (r = 0.817). Some cases had a disparity regarding the position of the sesamoids between the tangential view and the AP view due to misclassification on the AP view. We conclude that the scale of position of the sesamoid on the AP view is not valid in some cases, whereas the SRA is useful for assessing quantitatively the rotational position of the hallucal sesamoids in cases of hallux valgus.