Potential Use of Folate-polyethylene glycol (PEG)-Appended Dendrimer (G3) Conjugate with alpha-Cyclodextrin as DNA Carriers to Tumor Cells

We previously reported that polyamidoamine STARBURST dendrimer (generation 3, G3) (dendrimer) conjugate with alpha-cyclodextrin (alpha-CyD) having an average degree of substitution of 2.4 of alpha-CyD (alpha-CDE) provided remarkable aspects as novel carriers for DNA and siRNA. To develop novel alpha-CDE derivatives with tumor cell specificity, we prepared folate-appended alpha-CDEs (Fol-alpha-CDEs) and folate-polyethylene glycol (PEG)-appended alpha-CDEs (Fol-PalphaCs) with the various degrees of substitution of folate (DSF), and evaluated in vitro and in vivo gene transfer activity, cytotoxicity, cellular association and physicochemical properties. In vitro gene transfer activity of Fol-alpha-CDEs (G3, DSF 2, 5 or 7) was lower than that of α-CDE (G3) in KB cells, folate receptor (FR)-overexpressing cancer cells. Of the three Fol-PalphaCs (G3, DSF 2, 5 or 7), Fol-PalphaC (G3, DSF 5) had the highest gene transfer activity in KB cells. The activity of Fol-PalphaC (G3, DSF 5) was significantly higher than that of alpha-CDE (G3) in KB cells, but not in A549 cells, FR-negative cells. Negligible cytotoxicity of the pDNA complex with Fol-PalphaC (G3, DSF 5) was observed in KB cells or A549 cells up to a charge ratio of 100/1 (carrier/pDNA). The cellular association of the pDNA complex with Fol-PalphaC (G3, DSF 5) could be mediated by FR on KB cells, resulting in its efficient cellular uptake. Fol-PalphaC (G3, DSF 5) had higher binding affinity with folate binding protein (FBP) than alpha-CDE (G3), although the physicochemical properties of pDNA complex with Fol-PalphaC (G3, DSF 5) were almost comparable to that with alpha-CDE (G3), although the onset charge ratio and the compaction ability of Fol-PalphaC (G3, DSF 5) were slightly different. Fol-PalphaC (G3, DSF 5) tended to show higher gene transfer activity than alpha-CDE (G3) 12 h after intratumoral administration in mice. These results suggest that Fol-PalphaC (G3, DSF 5), not Fol-alpha-CDEs, could be potentially used as a FR-overexpressing cancer cell-selective DNA carrier

ゲンパツ フメイ ガン ニオケル PET/CT ケンサ ノ ユウヨウセイ ニツイテ

We reported the utility of１８F-FDG-PET/CT examination for patients with cancer of unknownprimary origin. Twenty six patients（13 men, 13 women, aged 27-91 years, mean 71）were examined.The indication for PET/CT examination was tumor maker elevation（14 patients）, suspectedmetastatic tumor（14）and metastasis diagnosed histopathologically（3）. Patients weretold not to eat for at least four hours and a PET/CT image was obtained one hour after theadministration of 3.7MBq/kg FDG. From April to August 2006, 33 patients diagnosed with a cancerof unknown primary origin were referred to our hospital for PET/CT examination from anoutside institution. Twenty six patients could be investigated for outcomes. Seventeen patientsshowed an abnormal accumulation, with 14 of the 17 having their primary regions detected histopathologicallyor clinically. For one patient, the abnormal accumulation could not be determined toshow the origin. For 2 patients, it was difficult to diagnose if these abnormal accumulationsshowed the primary region or not, but CT examinations were helpful for a diagnosis. Seven of the9 patients who showed no abnormal accumulation were treated conservatively and the primaryregion for their cancer could not be detected during the follow up study. In 21 of 26 patients, theseresults were useful to select an appropriate therapy to be applied or a relevant examination. Weconsidered PET/CT examination, where it is possible to scan the whole body at one time, was veryuseful to get both morphologic and metabolic information. PET/CT examination showed a highersensitivity for detecting abnormal lesions than other imaging modalities

Development and Evaluation of Quantitative Immunoglobulin G Enzyme-Linked Immunosorbent Assay for the Diagnosis of Coronavirus Disease 2019 Using Truncated Recombinant Nucleocapsid Protein as Assay Antigen

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). Real-time RT-PCR is the most commonly used method for COVID-19 diagnosis. However, serological assays are urgently needed as complementary tools to RT-PCR. Hachim et al. 2020 and Burbelo et al. 2020 demonstrated that anti-nucleocapsid(N) SARS-CoV-2 antibodies are higher and appear earlier than the spike antibodies. Additionally, cross-reactive antibodies against N protein are more prevalent than those against spike protein. We developed a less cross-reactive immunoglobulin G (IgG) indirect ELISA by using a truncated recombinant SARS-CoV-2 N protein as assay antigen. A highly conserved region of coronaviruses N protein was deleted and the protein was prepared using an E. coli protein expression system. A total of 177 samples collected from COVID-19 suspected cases and 155 negative control sera collected during the pre-COVID-19 period were applied to evaluate the assay’s performance, with the plaque reduction neutralization test and the commercial SARS-CoV-2 spike protein IgG ELISA as gold standards. The SARS-CoV-2 N truncated protein-based ELISA showed similar sensitivity (91.1% vs. 91.9%) and specificity (93.8% vs. 93.8%) between the PRNT and spike IgG ELISA, as well as also higher specificity compared to the full-length N protein (93.8% vs. 89.9%). Our ELISA can be used for the diagnosis and surveillance of COVID-19

Evaluation of a Triage Checklist for Mild COVID-19 Outpatients in Predicting Subsequent Emergency Department Visits and Hospitalization during the Isolation Period: A Single-Center Retrospective Study

Managing mild illness in COVID-19 and predicting progression to severe disease are concerning issues. Here, we investigated the outcomes of Japanese patients with mild COVID-19, and identified triage risk factors for further hospitalization and emergency department (ED) visits at a single tertiary hospital. A triage checklist with 30 factors was used. Patients recommended for isolation were followed up for 10 days for subsequent ED visits or hospital admission. Overall, 338 patients (median age, 44.0; 45% women) visited the clinic 5.0 days (median) after symptom onset. Thirty-six patients were immediately hospitalized following triage; others were isolated. In total, 72 non-hospitalized patients visited the ED during their isolation, and 30 were hospitalized after evaluation for oxygen desaturation. The median ED visit and hospitalization durations after symptom onset were 5.0 and 8.0 days, respectively. The checklist factors associated with hospitalization during isolation were age > 50 years, body mass index > 25 kg/m2, hypertension, tachycardia with pulse rate > 100/min or blood pressure > 135 mmHg at triage, and >3-day delay in hospital visit after symptom onset. No patients died. Altogether, 80% of patients with mild COVID-19 could be safely isolated at home. Age, BMI, underlying hypertension, date after symptom onset, tachycardia, and systolic blood pressure at triage might be related to later hospitalization

A Practical Report on Follow-up Training of Pre-Graduation Nursing Skills in the COVID-19 Pandemic

COVID-19流行前と比べ,臨地実習の場で過ごす時間や経験が少ない状況下で臨床現場へと就職する学生たちの不安は計り知れず,このような不安は,リアリティショックや職場不適応,早期離職へとつながることが懸念される.そこで,卒業を間近に控えた学生たちを対象に「卒業前看護技術フォローアップ研修」を実施した.また,参加した学生たちからの評価や取り組みに対する意見を調査し,今後のフォローアップ研修内容の改善や研修内容の充実について検討した.結果として,「卒業前フォローアップ研修」全体および各看護技術項目については,満足度の高い評価を得ることができた.特に,身体侵襲を伴う看護技術や,就職後の早期から必要となる看護技術の習得に対する意識や期待感が高いことが伺えた.また「慣れない人との看護実践の経験」は,卒業前フォローアップ研修として効果的であり,「楽しさ」「新鮮さ」「緊張感」「学びの充足」につながると考えられた.(著者抄録)Compared to the situation before the COVID- 19 pandemic, students who start working in clinical settings with less time and experience spent in the field are immensely anxious. Such anxiety may lead to reality shock, workplace maladjustment, and early retirement. A “follow- up training for pre- graduation nursing skills” was conducted for students about to graduate. In addition, the evaluation and opinions of the students who participated in the training were surveyed, and future improvements and enhancement of the content of the follow- up training are discussed. As a result, the overall follow- up training before graduation and each nursing skill item received high student satisfaction ratings. In particular, they showed a high level of awareness and expectation for learning nursing skills that involve physical invasion and nursing skills that are necessary from the early stage after employment. In addition, the “experience of practicing nursing with unfamiliar people” is considered effective as follow- up training before graduation, leading to “enjoyment,” “freshness,” “tension,” and “fulfillment of learning.”departmental bulletin pape

〈Original〉 Comparison with regional glucose metabolism, amyloid deposition and tau deposition between dementia with Lewy bodies and Alzheimer disease brains

[Abstract] Purpose: Although some dementia with Lewy bodies (DLB) patients have Alzheimer disease (AD) pathology, there are no reports comparing AD and DLB by amyloid PET and tau PET in the same patient. The aim of this study was to investigate the regional difference of metabolic reduction, amyloid deposition and tau deposition between DLB and AD brain.Methods: Five consecutive subjects with DLB and five age and Mini-mental state examination score matched AD patients, who underwent both fluoro-2-deoxyglucose (FDG)-PET, Pittsburgh compound B (PiB)-amyloid PET and THK5351 (THK)-tau PET, were included in this study. Voxel-based statistical analysis was performed with Statistical Parametric Mapping for comparison with FDG, amyloid and tau accumulation between the DLB and AD brains.Results: FDG PET demonstrated that occipital metabolism in DLB group was significantly lower than that in AD group. PiB PET demonstrated that in the AD group the anterior to posterior cingulate cortices, supplementary motor area, and thalamic amyloid depositions were significantly higher than those in the DLB group. THK PET demonstrated that in the AD group the posterior cingulate cortices, posterior internal capsules, thalamic and peri-brainstem regions tau depositions were significantly higher than those in the DLB group.Conclusions: In the DLB brain, it is suggested that decreased regional glucose metabolism is unrelated to amyloid and tau depositions, and the amyloid and tau depositions in the cingulate gyri and thalamic regions in the AD brain are higher than those in DLB brain