認知症高齢者グループホーム職員の看取り体験の思い

This study explored caregiver perceptions related to experiences of end-of-life (EOL) care at group homes (GH) for elderly people with dementia. A self-administered questionnaire was designed to elicit GH staff perceptions by asking what they feel about their EOL care experiences at GH. Eighty one participants shared their perspectives. Results were classified into seven categories, “Anxiety and tension related to EOL care,” “Regret and condolence,” “Response of care provided,” “Respect for deceased people,” “Pride in providing EOL care,” “Creative view of life and death,” and “Familial sense of loss,”. Although staff members who experienced EOL care felt imperfect, they were satisfied with the care they provided. Moreover, they had positive views in terms of EOL care experiences, which seemed to engender their personal growth. The study also revealed a distinctive view of EOL care in GH, where caregivers felt as if they had lost a family member when they confronted a resident’s death

青森県民の健康寿命アップ対策としての「心疾患10年リスク」の活用について(青森県の健康寿命アップと保健大学の取り組み, 第2回青森県立保健大学学術研究集会)

publisher青森市国立情報学研究所の「学術雑誌公開支援事業」により電子化されました

The High Content of Ent-11α-hydroxy-15-oxo-kaur- 16-en-19-oic Acid in Adenostemma lavenia (L.) O. Kuntze Leaf Extract: With Preliminary in Vivo Assays

Background: Ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic acid (11αOH-KA) is a multifunctional biochemical found in some ferns, Pteris semipinnata, and its congeneric species. Although a number of therapeutic applications of 11αOH-KA have been proposed (e.g., anti-cancer, anti-inflammation, and skin whitening), the content of 11αOH-KA in these ferns is not high. Adenostemma lavenia (L.) O. Kuntze, an Asteraceae, has also been reported to contain 11αOH-KA. The decoction (hot water extract) of whole plants of A. lavenia is used as a folk remedy for inflammatory disorders, such as hepatitis and pneumonia, suggesting that 11αOH-KA may be the ingredient responsible for the medicinal properties of this plant. Methods: The anti-melanogenic activities of the water extracts of A. lavenia leaves and Pteris dispar Kunze (a cognate of P. semipinnata) leaves were compared in mouse B16F10 melanoma cells. The amount of 11αOH-KA was measured by using liquid chromatography spectrometry. C57BL/6J mice were treated with the water extract of A. lavenia leaf, and the blood concentration of 11αOH-KA was measured. The in vivo efficacy of the water extract of A. lavenia leaf was evaluated according to tis anti-melanogenic activity by monitoring hair color. Results: Although both the extracts (A. lavenia and P. dispar Kunze) showed high anti-melanogenic activities, only A. lavenia contained a high amount of 11αOH-KA, approximately 2.5% of the dry leaf weight. 11αOH-KA can be purified from A. lavenia leaves in two steps: water extraction followed by chloroform distribution. The treatment of mice with the water extract of A. lavenia leaf suppresses pigmentation in their hairs. Conclusions: Despite the small number of mice examined, the present preliminary result of the suppressed hair pigmentation suggests that the water extract of A. lavenia leaf and the ingredient that is possibly responsible for this—11αOH-KA—are new materials for oral cosmetics. The results may also be helpful in the future development of functional foods and methods to treat patients suffering from hyperpigmentation disorders, such as melasma

CRISPR screening identifies M1AP as a new MYC regulator with a promoter-reporter system

Background MYC is one of the proto-oncogenes contributing to tumorigenesis in many human cancers. Although the mechanism of MYC regulation is still not fully understood, learning about the comprehensive mechanism controlling the transcriptional activity of MYC will lead to therapeutic targets. The CRISPR/Cas9 library system is a simple and powerful screening technique. This study aims to identify new transcriptional upstream activators of MYC using the CRISPR activation library with new promoter-reporter systems. Methods and Results The MYC promoter-reporter system was developed with a photoconvertible fluorescent protein, Dendra2, and named “pMYC-promoter-Dendra2.” This MYC promoter-reporter system was designed to harbor a proximal MYC promoter at (3.1 kb). Both the CRISPR activation library and pMYC-promoter-Dendra2 were induced to HEK 293T cells, and Dendra2-positive cells, that are supposed that MYC should be upregulated, were collected individually by a cell sorter. Among the 169 cells collected, 12 clones were successfully established. Then, pMYC-promoter-Dendra2 was transfected again into these 12 clones, and two of 12 clones showed Dendra2 positivity. In this procedure, the cells with non-specific autofluorescence were correctly distinguished by utilizing the photoswitchable character of Dendra2. Using extracted genomic DNA of these two Dendra2 positive clones, polymerase chain reaction (PCR) was performed to amplify the guide RNA (gRNA) containing region, which was introduced by the CRISPR activation library. Eventually, PLEKHO2, MICU, MBTPS1, and M1AP were identified, and these gRNAs were transfected individually into HEK 293T cells again using the CRISPR activation system. Only M1AP gRNA transfected cells showed Dendra2-positive fluorescence. Then, the overexpression vector for M1AP with a doxycycline-inducible vector confirmed that M1AP induced high MYC expression by real-time quantitative PCR and western blot. Furthermore, the dual-luciferase assay showed a significant increase of promoter activity, and MYC mRNA was higher in M1AP- overexpressing cells. M1AP is highly expressed in several cancers, though, a positive correlation between M1AP and MYC was observed only in human acute myeloid leukemia. Conclusion The present study confirmed that the experimental method using the CRISPR library technology functions effectively for the identification of molecules that activate endogenous MYC. This method will help elucidate the regulatory mechanism of MYC expression, as well as supporting further drug research against malignant tumors

Efficient Identification of the MYC Regulator with the Use of the CRISPR Library and Context-Matched Database Screenings

MYC is a major oncogene that plays an important role in cell proliferation in human cancers. Therefore, the mechanism behind MYC regulation is a viable therapeutic target for the treatment of cancer. Comprehensive and efficient screening of MYC regulators is needed, and we had previously established a promoter screening system using fluorescent proteins and the CRISPR library. For the efficient identification of candidate genes, a database was used, for which mRNA expression was correlated with MYC using datasets featuring “Similar” and “Not exactly similar” contexts. INTS14 and ERI2 were identified using datasets featuring the “Similar” context group, and INTS14 and ERI2 were capable of enhancing MYC promoter activity. In further database analysis of human cancers, a higher expression of MYC mRNA was observed in the INTS14 mRNA high-expressing prostate and liver cancers. The knockdown of INTS14 in prostate cell lines resulted in decreased MYC mRNA and protein expression and also induced G0/1 arrest. This study confirmed that CRISPR screening combined with context-matched database screening is effective in identifying genes that regulate the MYC promoter. This method can be applied to other genes and is expected to be useful in identifying the regulators of other proto-oncogenes

Pre- and post-synaptic stimulation by nicotine and noradrenalin on the spontaneous beating of cardiomyocytes.

<p><b>A)</b> Effects of nicotine (1 µM). Left: representative recording of extracellular potentials before and 6 min after application of nicotine to a mono-culture of VMs treated for 5 days with GDNF 10 ng/ml ([VM+GDNF]), co-cultures of VMs/SNs treated with vehicle only ([VM/SN+Vehicle], and co-cultures treated 5 days with GDNF 10 ng/ml ([VM/SN+GDNF]). Right: percent changes of spontaneous beating rate from baseline. Values are means±SD (n = 4), *p<0.05 vs. VM+GDNF. <b>B)</b> Effects of noradrenalin (10 µM). Left: representative extracellular potentials of VMs before and 3 min after application of noradrenalin to a mono-culture of VMs/SNs treated for 5 days with GDNF 10 ng/ml ([VM+GDNF]), co-cultures of VMs/SNs treated with vehicle only ([VM/SN+Vehicle]), co-cultures treated 5 days with GDNF 10 ng/ml ([VM/SN+GDNF]). Right: percent changes of spontaneous beating rate from baseline. [VM+Vehicle] : mono-cultures of VMs treated with vehicle only, [VM+NGF]: mono-cultures of VMs treated 5 days with NGF 50 ng/ml, [VM+GDNF]: mono-cultures of VMs treated 5 days with GDNF 10 ng/ml, respectively. Values are means±SD (n = 4). *Significantly different from baseline in each group (p<0.05, n = 4); †p<0.05 vs the VM/SN group (n = 4); #p<0.05 vs the VM/SN+NGF group (n = 4).</p

Axon growth of SNs toward GDNF-expressing VMs.

<p>GDNF- or mock-transfected VMs were prepared by using adenovirus vectors (AdGDNF-VMs and Mock-VMs) and co-cultured with SNs at a close distance (∼1 mm). <b>A)</b> A bright field image of the triangular co-culture (day 5) (left) and its schematic illustration (right). The axons from SN grow predominantly toward AdGDNF-VMs. Bar indicate 1 mm. Video images (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0065202#pone.0065202.s005" target="_blank">Video S2</a>) are available online. <b>B)</b> Sequential time-lapsed images. The pictures were taken at 0, 20, 40, 80 and 160 hours after removal of the glass-ring separation. Bar indicate 1 mm. <b>C)</b> Representative immunolabeling images of neurofilament-M (NF-M, red), α-actinine (AA, green) and GDNF (blue). Left: AdGDNF-VMs close to SNs, Right: Mock-VMs close to SNs. The NFM-positive axons (red fibrous structure) distribute abundantly on AdGDNF-VMs (green+blue), whereas only scarcely on Mock-VMs (green). Bars indicate 20 µm. <b>D)</b> The fraction of axon (NFM-positive area) distributing on VMs (AA-positive area). Values are means±SD of 5 co-cultures in each group. *significantly different from mock-VMs.</p