One-step embedding method for maintaining orientation of pathological tissue specimens using agar thin films.

Pathological histology examination involves handling a variety of specimens that are cut according to regulations and placed in cassettes. Tissue fragments in the cassettes are then diagnosed after processing, embedding, thin sectioning, staining and other procedures using a processing machine. Maintaining tissue fragment order and orientation during these processes is important for accurate diagnosis. In this study, we present a method of maintaining tissue fragment order and orientation using a thin film of ultra-high-strength agar and evaluate its usefulness during tissue sectioning.Cassettes were prepared, each containing three pieces of porcine liver, and compared embedding time with and without agar thin films (ATFs). Embedding was performed by three medical laboratory scientists with different levels of experience.To enable one-step tissue sample embedding, ATFs were integrated with samples in the cassettes. This resulted in an average reduction of 6.22 s of embedding time per cassette compared with traditional embedding methods.Through the use of ATFs, tissue fragment order and orientation is maintained, and embedding process time shortened. Additionally, ATFs are easily prepared and stored in 10% neutral buffered formalin over extended periods, allowing for immediate use during sectioning. This method is ideal to implement in busy pathology laboratories.

Quantitative assessment of chest compression techniques on an infant manikin.

BACKGROUND: Current cardiopulmonary resuscitation (CPR) guidelines recommend the two-finger technique (TFT) of chest compression (CC) in infants for a single rescuer. We hypothesized that healthcare providers cannot achieve adequate CC depth with TFT, even if using real-time visual feedback (RVF). METHODS: This was a cross-over study, randomizing participants to perform three sets of 2-min continuous CC, comparing (i) TFT with RVF, (ii) the one-hand technique (OHT) without RVF, and (iii) OHT with RVF. A standard CPR trainer manikin of a 3-month-old infant and a monitor/defibrillator that displays and records the quantitative CC quality were used. We set a target compression depth of 40-50 mm and a target compression rate of 100-120/min. Data were analyzed using the Friedman test and Bonferroni correction. Statistical significance was defined as P-value of< 0.05. RESULTS: Fifty-nine healthcare providers participated in the study. The mean compression depth was 24 mm (interquartile range [IQR], 22-26 mm) in TFT with RVF and 43 mm (IQR, 38-48 mm) in OHT without RVF, P < 0.001. The proportion of adequate CC depth was 0% (IQR, 0-0%) in TFT with RVF, 22% (IQR, 5-54%) in OHT without RVF, and 62% (IQR, 29-83%) in OHT with RVF. The mean compression rate was within the target range in all three techniques. CONCLUSIONS: The TFT cannot produce the CC depth that meets the recommendation of the current CPR guidelines for an infant with RVF, whereas the OHT does.

Epidemiology of child mortality and challenges in child death review in Japan: The Committee on Child Death Review: A Committee Report: The Committee on Child Death Review: A Committee Report.

We performed a retrospective survey and verification of the medical records of death cases of children (and adolescents; aged <18 years) between 2014 and 2016 in pediatric specialty training facilities in Japan. Of the 2,827 registered cases at 163 facilities, 2,348 cases were included. The rate of identified deaths compared with the demographic survey, was 18.2%-21.0% by age group. The breakdown of deaths was determined as follows: 638 cases (27.2%) were due to external factors or unknown causes, 118 (5.0%) were suspected to involve child maltreatment, 932 (39.7%) were of moderate or high preventability or were indeterminable. Further detailed verification was required for 1,333 cases (56.8%). Comparison of the three prefectures with high rates of identified deaths in Japan revealed no significant differences, such as in the distribution of diseases, suggesting that there was little selection bias. The autopsy rate of deaths of unknown cause was 43.4%, indicating a high ratio of forensic autopsies. However, sufficient clinical information was not collected; therefore, thorough evaluations were difficult to perform. Cases with a moderate or high possibility of involvement of child maltreatment accounted for 5%, similar to previous studies. However, more objective evaluation is necessary. Preventable death cases including potentially preventable deaths accounted for 25%, indicating that proposals need to be made for specific preventive measures. Individual primary verification followed by secondary verification by multiple organizations is effective. It is anticipated that a child death review (CDR) system with such a multi-layered structure will be established; however, the following challenges were revealed: The subjects of CDR are all child deaths. Even if natural death cases are entrusted to medical organizations, and complicated cases to other special panels, the numbers are very high. Procedures need to be established to sufficiently verify these cases. Although demographic statistics are useful for identifying all deaths, care must be taken when interpreting such data. Detailed verification of the cause of death will affect the determination of subsequent preventability. Verification based only on clinical information is difficult, so a procedure that collates non-medical information sources should be established. It is necessary to organize the procedures to evaluate the involvement of child maltreatment objectively and raise awareness among practitioners. To propose specific preventive measures, a mechanism to ensure multiprofessional diverse perspectives is crucial, in addition to fostering the foundation of individual practitioners. To implement the proposed measures, it is also necessary to discuss the responsibilities and authority of each organization. Once the CDR system is implemented, verification of the system should be repeated. Efforts to learn from child deaths and prevent deaths that are preventable as much as possible are essential duties of pediatricians. Pediatricians are expected to undertake the identified challenges and promote and lead the implementation of the CDR system. This is a word-for-word translation of the report in J. Jpn. Pediatr. Soc. 2019; 123 (11): 1736-1750, which is available only in the Japanese language.

Simultaneous Quantification of Ellagitannins and Related Polyphenols in Geranium thunbergii Using Quantitative NMR.

Compared to commonly employed liquid chromatography-based methods, quantitative nuclear magnetic resonance (qNMR) is a recently developed method for accurate quantification of natural compounds in extracts. The simultaneous quantification of ellagitannins and the related polyphenols of Geranium thunbergii were studied using qNMR after a short-term and long-term decoction. The qNMR fingerprint for quantifying ellagitannin was presented in this work. Geraniin was observed in the short-term decoction as a major component while corilagin was the major component of the long-term decoction. An aqueous acetone extract of G. thunbergii after long-term decoction was extracted with diethyl ether, ethyl acetate, and n-butanol. Corilagin was found as a major constituent in the ethyl acetate and n-butanol extracts. Furthermore, the contents of these polyphenols in G. thunbergii from six locations in Japan and three locations in China were quantified. The contents of geraniin and corilagin in G. thunbergii from Japan were higher than those from China. Our finding raised the possibility that qNMR can be effectively employed as a simple, accurate, and efficient method for quantification of ellagitannins in medicinal plants.

Effective Use of Computed Tomography for Detection of Press-Through-Package Sheet Ingestion in a Pediatric Patient.

Press-through-package sheet (PTPS) ingestion can cause perforation of gastrointestinal tract. Such incidents, which require immediate medical attention, are often observed in elderly people. In this report, we describe the case of a 12-year-old patient who ingested PTPS. The patient, who has attention-deficit/hyperactivity disorder, presented with abdominal pain at our hospital. While it was not revealed by a chest x-ray, computed tomography scan showed a PTPS lodged in his lower esophagus. The ingested PTPS was removed by endoscopy without complications. Press-through-package sheets ingestion can occur in pediatric patients, and computed tomography scan is useful in the diagnosis.

[A case of advanced rectal cancer treated effectively with intersphincteric resection and preoperative chemotherapy].

An approximately 50-year-old man with rectal cancer(RbP)[ cT3(cA), cN3, cM0, and cStage IIIb] who desired anus preservation was administered mFOLFOX6 therapy. This treatment decreased the size of both the tumor and the lymph node, and intersphincteric resection (ISR) was performed. Histopathology demonstrated tumor invasion beyond the muscularis propria, and the histological effectiveness was Grade 2. Because computed tomography showed an abscess in the dissection area, we performed postoperative drainage and the patient recovered. Therapy with mFOLFOX6 was repeated after the patient was discharged from hospital. Increased adoption of anus-preserving surgery is expected via successful control of local recurrence and distant metastasis by neoadjuvant chemotherapy.

High-throughput screening method for promoter activity using bead display and a ligase ribozyme.

In this report, we describe the development of a novel in vitro high-throughput system for detecting and screening promoter activity; the method employs emulsified reactions and a ligase ribozyme. In our study, a promoter DNA fragment containing the ribozyme gene was immobilized on a bead by using emulsion PCR, followed by in vitro transcription of the immobilized DNA in water-in-oil emulsions. Owing to the self-ligation activity of the ribozyme, it was co-transcriptionally linked to the active promoter immobilized on the beads. The bead complex containing the active promoter sequence was then labeled by reverse transcription with a fluorescently labeled primer. Employing flow cytometry, the fluorescence intensity corresponding to the strength of each promoter was observed, indicating the applicability of the system for promoter evaluation. Moreover, two rounds of screening with T7 RNA polymerase using a cell sorter enriched the T7 promoter fragment by 70 folds from a 1:100 mixture of T7 promoter and SP6 promoter fragments, suggesting that this system can be used to screen promoters.