Long-term storage under pressure in deep sea improved the microbiological safety and physical properties of whale meat.

This study aimed to clarify the effects of deep-sea pressure storage on the quality of whale meat, especially microbiological safety and physical properties, to examine the effectiveness of deep-sea storage for long-term aging of whale meat. Microbiological safety, physical properties, color and appearance, water content, water activity, and pH of whale meat were examined after storage in the deep sea at depths of 2200-6000 m (22-60 MPa) for 4 months. During storage under high pressure at a depth of >4000 m (40 MPa), the growth of aerobic bacteria was inhibited in whale meat. The toughness of whale meat stored in deep sea at a depth of >4000 m became significantly tender than that before deep-sea storage. Long-term storage of whale meat under high pressure and low-temperature conditions in the deep sea at a depth of >4000 m was clarified to improve the microbiological safety and tenderness of whale meat.

[Large-vessel vasculitis developed after use of long-acting granulocyte colony-stimulating factor in a patient with diffuse large B-cell lymphoma].

A 75-year-old man was diagnosed with diffuse large B-cell lymphoma originating from the paranasal sinuses. Curative induction chemotherapy was initiated and pegfilgrastim was administered on day5 of the first cycle as primary prophylaxis. The patient developed headache on day7 and fever on day11. These symptoms persisted despite treatment with antibiotics and antifungal agents. Computed tomography (CT) after admission revealed wall thickening in the aortic arch. Chest contrast-enhanced CT also revealed contrast enhancement in the thickened aortic wall. Results of blood cultures and serological tests for autoantibodies were negative, indicating that the clinical manifestations were not due to infection or a specific collagen disease. The final diagnosis was drag-induced large vessel vasculitis induced by long-acting granulocyte colony-stimulating factor (G-CSF). The patient's symptoms and large-vessel wall thickening immediately resolved after treatment with a glucocorticoid (prednisolone, 0.6 mg/kg/day). Aortitis should be considered as a differential diagnosis when fever is observed in a patient who received long-acting G-CSF during chemotherapy.

Cold-Restraint Stress-Induced Ultrasonic Vocalization as a Novel Tool to Measure Anxiety in Mice.

Ultrasonic vocalization (USVs) is a promising tool to measure behavioral anxiety in rodents as USV recording is noninvasive, behaviorally relevant, ethological, and reproducible. Studies reporting the effects of stress-induced USVs in adult mice remain limited and debated. We investigated the conditions under which mice emit aversive USVs and evaluated the effects of psychiatric drugs on stress-induced USVs. Male C57BL/6J mice were used. USVs during entire stress sessions were recorded according to their frequency. To investigate the effect of psychiatric drugs on USVs, the number of USVs under cold-restraint stress conditions before and after drug administration was compared. Immediately after stress exposure, blood samples were collected and plasma corticosterone levels were measured. The combination of cold and restraint stress conditions significantly increased the USV numbers and plasma corticosterone levels compared with each stress alone. A benzodiazepine anxiolytic (midazolam) and δ-opioid receptor agonist putative anxiolytic (KNT-127) significantly reduced the stress-induced USV number and plasma corticosterone levels; however, a monoaminergic antidepressant (duloxetine) and N-methyl-D-aspartic acid receptor antagonist antidepressant (ketamine) did not reduce the USV numbers. No changes were noted in the USV numbers after repeated exposure to cold-restraint stress on days 1 and 3. The suppressive effect of midazolam on day 3 was comparable to that on day 1. Stress-induced USV may be used as a quantitative measure of anxiety to systematically assess the effects of anxiolytics. Therefore, cold-restraint stress-induced USVs may be used as a novel tool to measure rodent anxiety and as a useful anxiolytic-screening system.

An infectivity-enhancing site on the SARS-CoV-2 spike protein targeted by antibodies.

Antibodies against the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein prevent SARS-CoV-2 infection. However, the effects of antibodies against other spike protein domains are largely unknown. Here, we screened a series of anti-spike monoclonal antibodies from coronavirus disease 2019 (COVID-19) patients and found that some of antibodies against the N-terminal domain (NTD) induced the open conformation of RBD and thus enhanced the binding capacity of the spike protein to ACE2 and infectivity of SARS-CoV-2. Mutational analysis revealed that all of the infectivity-enhancing antibodies recognized a specific site on the NTD. Structural analysis demonstrated that all infectivity-enhancing antibodies bound to NTD in a similar manner. The antibodies against this infectivity-enhancing site were detected at high levels in severe patients. Moreover, we identified antibodies against the infectivity-enhancing site in uninfected donors, albeit at a lower frequency. These findings demonstrate that not only neutralizing antibodies but also enhancing antibodies are produced during SARS-CoV-2 infection.

[The Strength of the Sense of Shame of the Aged in Care Scenes: Comparison Between Whether or not There is Anyone Else Around Them].

Medical staff in a hospital or nursing facility should take care of aged individuals with dignity and respect. We conducted a survey on aged individuals to derive under what care circumstances they had a sense of shame, using 12 illustrations, drawn by ourselves, which were common daily care scenes where nurses and patients meet. This survey was conducted at 4 care facilities in A prefecture, Japan. The number of surveyed persons was 43, with the following exclusion criteria: over 60 years old, more than third level of care needed, and non suspected of having dementia. We got the following results from the answers of 41 persons: 1. When elder persons are surrounded by people other than the care staff, they feel more of a sense of shame than when alone; 2. They feel more sense of shame when they use a wheelchair than when they use crutches; 3. They do not feel much shame when they get a bed-bath, even if other persons are there; and 4. Male patients feel more shame than females when they meet their family. These results suggest that elderly patients feel a stronger sense of shame when they are seen by others than when they are seen by care staff. The result 2 suggests that the use of a wheelchair exposes their physical weakness to others. Males feel a stronger sense of shame when they show a weakness in their gender role. We conclude that the sense of shame of aged individuals in daily life scenes in a care facility depends on their gender and whether or not they are surrounded by other persons.

A TCR-like antibody against a proinsulin-containing fusion peptide ameliorates type 1 diabetes in NOD mice.

Type 1 diabetes (T1D) is an autoimmune disease caused by destruction of insulin-producing ß cells. The response of autoreactive T cells to ß cell antigens plays a central role in the development of T1D. Recently, fusion peptides composed by insulin C-peptide fragments and other proteins were reported as ß cell target antigens for diabetogenic CD4+ T cells in non-obese diabetic (NOD) mice. In this study, we generated a T cell-receptor (TCR)-like monoclonal antibody (mAb) against a fusion peptide bound to major histocompatibility complex (MHC) class II component to elucidate the function of the fusion peptides in T1D. In addition, we developed a novel NFAT-GFP TCR reporter system to evaluate the TCR-like mAb. The NFAT-GFP reporter T cells expressing the diabetogenic TCR were specifically activated by the fusion peptide presented on the MHC class II molecules. By using the NFAT-GFP reporter T cells, we showed that the TCR-like mAb blocks the diabetogenic T cell response against the fusion peptide presented on the MHC class II molecules. Furthermore, the development of T1D was ameliorated when pre-diabetic NOD mice were treated with this mAb. These findings suggest that NFAT-GFP reporter T cells are useful to assess the function of specific TCR and the recognition of fusion peptides by T cells is crucial for the pathogenesis of T1D.

[Nursing Care Students' Image of Shame -Comparison Between First and Second Year Nursing Students].

It is not easy for nurses to estimate a patient's degree of shame, as the sense of shame depends on each person's personality, but nurses are requested to evaluate it as correctly as possible and to reduce the patient's mental load. We presume that most of the sense of shame is generated by body defects or disadvantages recognized by the patient. In this study, we tried to measure the degree of shame and to improve the basic nursing curriculum, depending on students' school year, under the assumption of what cases the nurses would frequently meet in a hospital. We prepared 13 figures that show common cases in hospitals. In these figures: 1) 6 figures show cases in which a nurse touched a patient's body; 2) 3 figures show common daily life; and 3) 4 figures show cases in which there are other people around the patient. A questionnaire was given to the first and second year students in A Nursing University, and we allocated scores of 1 to 10: 1 is "no-shame", and 10 is "very much shame". The students answered that patients must feel shame when: 1) they take off their clothes, 2) they show their disability to another person even without taking their clothes off, and 3) having people other than medical staff around them. In the results, as 2) appeared more strongly in the second year students than in the first, we thought that the second year students could surmise a patient's position in a hospital through the effect of the nursing education.

C/EBPß regulates Vegf gene expression in granulosa cells undergoing luteinization during ovulation in female rats.

The ovulatory LH-surge increases Vegf gene expression in granulosa cells (GCs) undergoing luteinization during ovulation. To understand the factors involved in this increase, we examined the roles of two transcription factors and epigenetic mechanisms in rat GCs. GCs were obtained from rats treated with eCG before, 4 h, 8 h, 12 h and 24 h after hCG injection. Vegf mRNA levels gradually increased after hCG injection and reached a peak at 12 h. To investigate the mechanism by which Vegf is up-regulated after hCG injection, we focused on C/EBPß and HIF1α. Their protein expression levels were increased at 12 h. The binding activity of C/EBPß to the Vegf promoter region increased after hCG injection whereas that of HIF1α did not at this time point. The C/EBPß binding site had transcriptional activities whereas the HIF1α binding sites did not have transcriptional activities under cAMP stimulation. The levels of H3K9me3 and H3K27me3, which are transcriptional repression markers, decreased in the C/EBPß binding region after hCG injection. The chromatin structure of this region becomes looser after hCG injection. These results show that C/EBPß regulates Vegf gene expression with changes in histone modifications and chromatin structure of the promoter region in GCs undergoing luteinization during ovulation.

Invariant chain p41 mediates production of soluble MHC class II molecules.

Major histocompatibility complex class II (MHC II) molecules are mainly expressed on antigen presentation cells and play an important role in immune response. It has been reported that MHC II molecules are also detected in serum as a soluble form (sMHC II molecules), and they are considered to be involved in the maintenance of self-tolerance. However, the mechanism by which sMHC II molecules are produced remains unclear. Invariant chain (Ii), also called CD74, plays an important role in antigen presentation of MHC II molecules. In the present study, we analyzed the role of Ii on the production of sMHC II molecules. We found that the amount of sMHC II molecules in serum was decreased in Ii-deficient mice compared to wild-type mice. sMHC II molecules were secreted from cells transfected with MHC II molecules and Ii but not from cells transfected with MHC II molecules alone. Moreover, isoform p41 of Ii-transfected cells induced more sMHC II molecules compared to isoform p31-transfected cells. The molecular weight of sMHC II molecules from MHC II and Ii p41-transfected cells was approximately 60 kDa, indicating that sMHC II molecules are a single heterodimer of α and ß chains that is not associated with micro-vesicles. From the analysis of Ii-deletion mutants, we found that the luminal domain of Ii p41 is crucial for the production of sMHC II molecules. These results suggested that Ii has an important role in production of sMHC II molecules.

Nursing Education Trial Using a Virtual Nightingale Ward.

Nursing department students are expected to correctly grasp the entire concept of nursing through their education. The authors created a movie of a Nightingale ward (virtual ward, hereafter) with an architectural computer design software for education. The students' reaction to the virtual ward was categorized into three viewpoints: that of nurses, of patients, and of nurses and patients in common. Most of the reactions in each viewpoint were: "easy to observe patients" in the nurses' viewpoint; "no privacy" in the patients' viewpoint; and "wide room" in the common viewpoint, respectively. These reactions show the effectiveness of using a virtual ward in nursing education. Because these reactions are characteristics of a Nightingale ward, and even students, who have generally less experiences, recognized these characteristics from the both viewpoints of nurses and patients.

Development of Teaching Materials in Basic Nursing Using Virtual Reality －Ceiling Image Teaching Materials of a Hospital Room－.

It is difficult to understand nursing practically for students who have just started studying nursing, because they cannot imagine the actual medical scene. The authors expect that they can improve nursing learning through experiences in virtual medical environments supported by ICT (information and commnication technology). We developed teaching materials in nursing education, and in a previous study we reported that the use of a virtual hospital was useful for students who had not had medical experience. The aim of the present study was to have students consider the meaning of a hospital room ceiling. First, we showed them a few ceiling patterns, and they selected the best one among the patterns. After that, we showed the same one in a virtual hospital room. They had negative opinions about what they had thought was the best one at this time. This demonstrates that the virtual hospital room is effective when students evaluate the ceiling as a part of the hospital room.

Sialic Acids on Varicella-Zoster Virus Glycoprotein B Are Required for Cell-Cell Fusion.

Varicella-zoster virus (VZV) is a member of the human Herpesvirus family that causes varicella (chicken pox) and zoster (shingles). VZV latently infects sensory ganglia and is also responsible for encephalomyelitis. Myelin-associated glycoprotein (MAG), a member of the sialic acid (SA)-binding immunoglobulin-like lectin family, is mainly expressed in neural tissues. VZV glycoprotein B (gB) associates with MAG and mediates membrane fusion during VZV entry into host cells. The SA requirements of MAG when associating with its ligands vary depending on the specific ligand, but it is unclear whether the SAs on gB are involved in the association with MAG. In this study, we found that SAs on gB are essential for the association with MAG as well as for membrane fusion during VZV infection. MAG with a point mutation in the SA-binding site did not bind to gB and did not mediate cell-cell fusion or VZV entry. Cell-cell fusion and VZV entry mediated by the gB-MAG interaction were blocked by sialidase treatment. N-glycosylation or O-glycosylation inhibitors also inhibited the fusion and entry mediated by gB-MAG interaction. Furthermore, gB with mutations in N-glycosylation sites, i.e. asparagine residues 557 and 686, did not associate with MAG, and the cell-cell fusion efficiency was low. Fusion between the viral envelope and cellular membrane is essential for host cell entry by herpesviruses. Therefore, these results suggest that SAs on gB play important roles in MAG-mediated VZV infection.

[Nursing Education Utilizing Experiences in a Virtual Hospital Ward].

Environmental design should be required at medical facilities for conducting medical practice safely and for making hospitalization comfortable. Many medical nursing students cannot imagine medical facilities, especially hospital wards, when they study medical environments in a basic nursing lecture. As a result, they cannot connect well with patient assistance. We employed a computer assisted designing software, "3D My Home Designer" (Mega Soft Company) that runs on Windows 8, and considered the usefulness of it for lectures on environmental design showing how to design a hospital ward for patients' optimal hospital stay. We drew a medical facility in 2-D first, transformed it into 3D images, and then created movies of a virtual hospital ward in which a patient walked around. These movies consisted of 3 kinds: a) hospital room with changeable wall color, b) different allocations of hospital room and nurse station, and c) a blurred ward which corresponded to how a patient with poor eyesight (cataract) would see a ward. We prepared as controls: a') still images of a hospital room, b') still images of ward, and c') a documentation on how a ward is seen by a patient with a cataract. We gave a questionnaire to students and nurses about these movies and still images (controls). In a) and b), there were no differences between the movies and still images in both students and nurses. In c), both students and nurses had a viewpoint from the patient with poor eyesight. From these results, we consider that the students, who have fewer experiences in a hospital, may understand the environments well by movies and the application of a virtual movie ward to nursing education may be useful in a lecture, depending on the readiness of the students.

Transport of misfolded endoplasmic reticulum proteins to the cell surface by MHC class II molecules.

Nascent MHC class II molecules are associated with the invariant chain and are transported to the endolysosomal pathway, where MHC class II molecules acquire peptide antigens. On the other hand, misfolded endoplasmic reticulum (ER) proteins are generally degraded in the cells and are neither expressed on the cell surface nor secreted. Here, we found that MHC class II molecules associate with some misfolded ER proteins via the peptide-binding groove in competition with invariant chain. The misfolded proteins associated with MHC class II molecules are transported intact to the cell surface without processing to peptides. Furthermore, these complexes efficiently stimulate antigen-specific B cells. These findings reveal that MHC class II molecules function as a chaperone for the cell surface expression of misfolded ER proteins. In addition, we suggest that MHC class II molecules present not only peptides but also intact host-cell-derived proteins on the cell surface. These findings provide new insights into the function of MHC class II molecules.

Structure-activity relationship of anthelmintic cyclooctadepsipeptides.

The relationship between cyclooctadepsipeptides and their anthelmintic efficacy was examined by converting the natural products, PF1022A, PF1022E and PF1022H. Some analogues substituted at the para position of the phenyllactate moiety showed higher or equivalent activity against the parasitic nematode, Ascaridia galli in chicken when compared with the parent compounds. It is suggested that lipophilicity and the polar surface area, in addition to structural requirements of the derivatives, influenced the anthelmintic efficacy in vivo.