Oral cancer diagnosis via a ferrocenylnaphthalene diimide-based electrochemical telomerase assay.

BACKGROUND: Telomerase is regarded as a good marker for cancer because it is unregulated in most tumor cells compared with normal cells. We evaluated telomerase activity in the lysate of tumor tissue and surrounding cells of oral cancer patients by an electrochemical technique, dubbed the electrochemical telomerase assay (ECTA). METHODS: The assay used ferrocenylnaphthalene diimide (FND) as the probe. Electrochemical telomerase substrate (ETS) primer immobilized on the electrode was elongated by telomerase and FND bound to the product to give rise to a current. The data were standardized with the change in current before and after the elongation, respectively. RESULTS: The change in current increased more than 30% in biopsy samples from most cancer patients, whereas the increase was 20% or lower in most healthy individuals. On the basis of this difference individual clinical samples were judged telomerase positive, ambiguous, or negative. The positive rates in the cancerous tissues and exfoliated cells (EOCs) of the patients were 85% and 90%, respectively, whereas the corresponding values were 50% and 10% by the telomerase repeat amplification protocol. Furthermore, the positive rate for the ECTA was 100% in early tumors smaller than 2 cm, and 95% and 82% of biopsy and exfoliated cells of healthy individuals were correctly judged as negative. Fifty-six unknown samples with EOCs tested were correctly judged to be cancerous or normal in 84% of cases. CONCLUSIONS: The ECTA yielded high hit rates for cancerous and normal cells, especially in EOCs, results indicating that this minimally invasive test is suitable for oral cancer diagnosis.

[Development of a One-minute Educational Video for Eye-drop Instillation and Changes in Instillation Behavior after Watching the Video].

We created a one-minute video titled "a simple method of eye-drop instillation" (video) for online instillation guidance, to compare the instillation method before and after study participants watch the video and verify the usefulness of watching the video. Moreover, we prepared a document questionnaire to investigate instillation habits and clarify instillation behavior. Study participants were randomly recruited from among students and faculty members via a poster posted at Tokushima Bunri University. The instillation behavior of the study participants was videotaped before and after they watched the video created by the authors. The images were played in a super slow motion, to confirm success or failure in instillation, drop sites, and eye-opening method. Of the 109 participants in the study, the successful instillation rate before and after watching the video was 55.0% and 69.7%, respectively. The use rate of wet wipes for finger disinfection before instillation increased from 0.0% before watching the video to 74.3% after watching the video. After watching the video, the blinking rate after instillation decreased from 95.4 to 45.0%, the rate of pressing the nasolacrimal duct increased from 2.8 to 77.1%, and the rate of wiping the drug solution spilled around the eyes increased from 89.9 to 98.2%. According to the questionnaire, 72.5% of the participants instilled one drop, 22.0% instilled two drops, and 5.5% instilled three drops or more. Watching the video significantly increased the successful instillation rate and improved instillation behavior. Thus, the video created by the authors can be used for online instillation guidance.

High-throughput cryopreservation of plant cell cultures for functional genomics.

Suspension-cultured cell lines from plant species are useful for genetic engineering. However, maintenance of these lines is laborious, involves routine subculturing and hampers wider use of transgenic lines, especially when many lines are required for a high-throughput functional genomics application. Cryopreservation of these lines may reduce the need for subculturing. Here, we established a simple protocol for cryopreservation of cell lines from five commonly used plant species, Arabidopsis thaliana, Daucus carota, Lotus japonicus, Nicotiana tabacum and Oryza sativa. The LSP solution (2 M glycerol, 0.4 M sucrose and 86.9 mM proline) protected cells from damage during freezing and was only mildly toxic to cells kept at room temperature for at least 2 h. More than 100 samples were processed for freezing simultaneously. Initially, we determined the conditions for cryopreservation using a programmable freezer; we then developed a modified simple protocol that did not require a programmable freezer. In the simple protocol, a thick expanded polystyrene (EPS) container containing the vials with the cell-LSP solution mixtures was kept at -30 °C for 6 h to cool the cells slowly (pre-freezing); samples from the EPS containers were then plunged into liquid nitrogen before long-term storage. Transgenic Arabidopsis cells were subjected to cryopreservation, thawed and then re-grown in culture; transcriptome and metabolome analyses indicated that there was no significant difference in gene expression or metabolism between cryopreserved cells and control cells. The simplicity of the protocol will accelerate the pace of research in functional plant genomics.

[Investigation of Successful Eyedrop Instillation Rates and Analysis of Drop Positions Using High-speed Digital Video Recording System].

We investigated the success rates of eyedrop instillation and the distance between the cornea and the dropper tip in 100 volunteers using high-speed digital video recording. Past eyedrop adherence studies assumed that instillation occurred without failure. The ideal distance between the cornea and dropper tip remained unclear, although the general estimate was approximately 2.54 cm (1 inch). This study was approved by the Institutional Review Boards of all participating medical institutions, and all volunteers provided written, informed consent. Successful instillation was defined as when 1 drop fell accurately into the eye on the first attempt. The instillation of ≥2 drops or drops delivered outside the eye was considered a failure. The distance between the eye and dropper tip was measured using still images from a paused digital video camera and a digital ruler. Forty percent of the volunteers instilled eyedrops without instructions from ophthalmologists, pharmacists, or other healthcare workers. When the images were analyzed, the success rate of the first instillation was 70.1%. When the eye was arbitrarily divided into 9 sections, most of the drop sites were the iris or the center of the eye. The distance between the dropper tip and cornea was 2.62±1.75 (median 2.20) cm. These results indicate that the generally recommended distance is usually followed. The successful instillation rate based on the distance from the dropper tip to the cornea was 77% at 1.6±0.88 cm and 54.9% at 4.8±1.25 cm.

The orbitofrontal cortex modulates parenting stress in the maternal brain.

Many mothers are adaptive, deploying successful coping strategies that mitigate the deleterious effects of parenting stress on caregiving, nevertheless, the neural mechanisms underlying these adaptive responses remain unclear. We utilized functional magnetic resonance imaging to investigate brain activity in 28 healthy mothers of typically developing, 2-to-3-year-old children in response to the feeding behavior of their own children versus that of other children. We then examined the correlation between maternal brain activation and subjective feelings of parenting stress. Brain regions associated with maternal motivation including the orbitofrontal cortex (OFC), ventral pallidum, periaqueductal gray (PAG), dorsal raphe nucleus (DRN), and anterior insular cortex (AIC)-as well as those associated with the recognition of one's own child's state (e.g., cerebellum)-exhibited significant activation in response to their own children. While mothers with higher activation in the OFC showed less parenting stress related to one's sense of competence in the parental role, mothers with higher co-activation of the OFC with both of the AIC and PAG/DRN, and with the cerebellum showed less parenting stress caused by child characteristics. Our findings suggest that well-balanced maternal brain mechanisms integrated by the OFC may provide effective adaptive responses in daily parenting scenarios.

Hg(OTf)2-catalyzed arylene cyclization.

Novel Hg(OTf) 2-catalyzed arylene cyclization was achieved with highly efficient catalytic turnover (up to 200 times). The reaction takes place via protonation of allylic hydroxyl group by in situ formed TfOH of an organomercuric intermediate to generate a cationic species. Subsequent smooth demercuration regenerates the catalyst.

Silent aspiration predicts mortality in older adults with aspiration pneumonia admitted to acute hospitals.

AIM: Aspiration pneumonia is a common disease resulting in poor outcomes for older adults. It is known that silent aspiration might lead to aspiration pneumonia, but its role in this is unclear. The aim of the present study was to investigate whether silent aspiration is associated with mortality in aspiration pneumonia patients. METHODS: A multicenter (acute hospital) study including 170 aspiration pneumonia patients was carried out. Cough latency time after inhalation of 1% citric acid mist was used to assess silent aspiration. Patients were divided into two groups: silent aspirator (SA) and no-SA groups based on the latency time of ≥29/<29 s. RESULTS: The mean age of the participants was 84.0 ± 8.8 years. A total of 82% of them had moderate aspiration pneumonia. A total of 99 (58.2%) patients were in the SA group. Silent aspiration patients were more likely to be inactive compared with those in the no-SA group (Barthel Index score 5 [IQR 0-45] vs 30 [IQR 5-65], P = 0.023) and had worse oral conditions (Oral Health Assessment Tool score 5.3 ± 3.2 vs 3.8 ± 3.1, P = 0.003). Nine of the SA patients and one of the no-SA patients (P = 0.047) died within a month. Multivariate regression analysis showed that silent aspiration was associated with mortality after adjusting for confounders (odds ratio 2.65, 95% CI 1.01-6.98, P = 0.049). CONCLUSIONS: Silent aspiration detected by a cough latency test can predict 1-month mortality in older patients with aspiration pneumonia. More studies are required to investigate whether enhancing cough reflex treatments would help patients recover from aspiration pneumonia. Geriatr Gerontol Int 2018; 18: 828-832.

Electrochemical telomerase assay for screening for oral cancer.

Telomerase has long been known to be a marker for cancer. We have developed a new method of detecting it: the electrochemical telomerase assay (ECTA). We have previously confirmed that the assay is easier to do and more precise than the conventional telomeric repeat amplification protocol, which is currently the most widely used. Here we describe a pilot study made to establish a screening system for oral cancer using ECTA. We evaluated three types of clinical samples obtained from 44 patients with oral cancer and 26 healthy volunteers: exfoliated cells from the whole oral cavity, exfoliated cells from local lesions, and tissue from the lesion itself. The current increase ratio (Δi) obtained by ECTA was significantly higher in the oral cancer group for each type of sampling used. The threshold value for Δi was 19% when calculated by analysis of receiver-operating characteristic curves. Sensitivity and specificity values were 86% and 85% for cells from the oral cavity, 82% and 85% in cells from local lesions, and 95% and 92% in cells from the tumour itself, respectively. There were also no significant differences in sensitivity and specificity associated with age, size of tumour, site of lesion, or degree of malignancy. ECTA therefore seems to be a promising assay for screening for oral cancer.