[Understanding public health nurses' experience, knowledge, and perception of bioterrorism training in Japan].

Objectives　Bioterrorism is a disease in which the attacks can be covert and latent, take time to manifest, and, when discovered, can result in large outbreaks. When detected, they can become large-scale outbreaks. Therefore, preparedness is essential for early detection and response for damage mitigation. Although public health nurses (PHNs) of public health centers are responsible for detecting and dealing with bioterrorism, their preparation status is not clear. Therefore, this study aimed to elucidate the status of training experience, knowledge, and awareness of bioterrorism among PHNs working in public health centers' infectious disease control departments in Japan's metropolitan areas.Methods　This was a cross-sectional, descriptive study. The participants were PHNs in the infectious disease control departments of 88 public health centers in Tokyo and its three neighboring prefectures. This are densely populated metropolitan areas with many mass gathering events and a high probability of bioterrorism. An anonymous, self-administered questionnaire survey was conducted by mail in 2019. Two PHNs per location were surveyed regarding their attributes, training experience, knowledge, and perceptions.Results　We received 71 responses (40.3%). A total of 10 (14.1%) PHNs reported having experience with bioterrorism training in the workplace. Regarding the knowledge of the four infectious diseases with a high probability of bioterrorism, more than 95% of the respondents answered they had heard of them. However, few were aware of related treatments or terrorist responses. Although survey respondents were aware of the seriousness of bioterrorism and the effectiveness of preparedness, they did not have sufficient opportunity or time to have training. They tended to be less confident in their response and were willing to have training.Conclusion　PHNs who responded to this survey do not have sufficient training experience, knowledge, opportunities, and time for bioterrorism training, and establishing a training system to improve bioterrorism preparedness is a challenge. It will be necessary for PHN students to learn about the existence of bioterrorism. Further, PHNs in public health centers need to be trained at least once using online services, including brushing up on information as needed. Based on the lessons learned from the coronavirus disease 2019 pandemic, health centers will formulate a Health Crisis Management Plan. This will be effective in fiscal 2024, and we believe that it is essential that preparedness against bioterrorism be a part of this plan.

Analysis of the Effectiveness of Metaheuristic Methods on Bayesian Optimization in the Classification of Visual Field Defects.

Bayesian optimization (BO) is commonly used to optimize the hyperparameters of transfer learning models to improve the model's performance significantly. In BO, the acquisition functions direct the hyperparameter space exploration during the optimization. However, the computational cost of evaluating the acquisition function and updating the surrogate model can become prohibitively expensive due to increasing dimensionality, making it more challenging to achieve the global optimum, particularly in image classification tasks. Therefore, this study investigates and analyses the effect of incorporating metaheuristic methods into BO to improve the performance of acquisition functions in transfer learning. By incorporating four different metaheuristic methods, namely Particle Swarm Optimization (PSO), Artificial Bee Colony (ABC) Optimization, Harris Hawks Optimization, and Sailfish Optimization (SFO), the performance of acquisition function, Expected Improvement (EI), was observed in the VGGNet models for visual field defect multi-class classification. Other than EI, comparative observations were also conducted using different acquisition functions, such as Probability Improvement (PI), Upper Confidence Bound (UCB), and Lower Confidence Bound (LCB). The analysis demonstrates that SFO significantly enhanced BO optimization by increasing mean accuracy by 9.6% for VGG-16 and 27.54% for VGG-19. As a result, the best validation accuracy obtained for VGG-16 and VGG-19 is 98.6% and 98.34%, respectively.

Physiological responses (Hsps 60 and 32, caspase 3, H2O2 scavenging, and photosynthetic activity) of the coral Pocillopora damicornis under thermal and high nitrate stresses.

This study explored the physiological responses of the coral Pocillopora damicornis to high nitrate concentrations and thermal stresses. The expression of heat shock proteins Hsp60 and Hsp32, Symbiodiniaceae density, Chl a concentration, Fv/Fm, H2O2 scavenging, and caspase 3 activity varied during 60 h incubations at 28 °C or 32 °C, ambient or high nitrate (~10 µM) concentrations, and their combinations. In combined stresses, corals showed a rapid and high oxidation level negatively affecting the Symbiodiniaceae density and Chl a concentration at 12 h, followed by caspase 3 and Hsps upregulations that induced apoptosis, bleaching and tissue detachment. Corals under thermal stress showed the highest oxidation and upregulation of Hsps and caspase 3 resulting in coral discoloration. High nitrate treatment alone did not seriously affect the coral function. Results showed that combined stress treatment severely affected coral physiology and, judging from the condition of detached tissues, these corals might have lower chances to recover.

Chlorophyll fluorescence - A tool to assess photosynthetic performance and stress photophysiology in symbiotic marine invertebrates and seaplants.

Chlorophyll a fluorescence is increasingly being used as a rapid, non-invasive, sensitive and convenient indicator of photosynthetic performance in marine autotrophs. This review presents the methodology, applications and limitations of chlorophyll fluorescence in marine studies. The various chlorophyll fluorescence tools such as Pulse-Amplitude-Modulated (PAM) and Fast Repetition Rate (FRR) fluorometry used in marine scientific studies are discussed. Various commonly employed chlorophyll fluorescence parameters are elaborated. The application of chlorophyll fluorescence in measuring natural variations, stress, stress tolerance and acclimation/adaptation to changing environment in primary producers such as microalgae, macroalgae, seagrasses and mangroves, and marine symbiotic invertebrates, namely symbiotic sponges, hard corals and sea anemones, kleptoplastic sea slugs and giant clams is critically assessed. Stressors include environmental, biological, physical and chemical ones. The strengths, limitations and future perspectives of the use of chlorophyll fluorescence technique as an assessment tool in symbiotic marine organisms and seaplants are discussed.

Xylan utilisation promotes adaptation of Bifidobacterium pseudocatenulatum to the human gastrointestinal tract.

Dietary carbohydrates impact the composition of the human gut microbiota. However, the relationship between carbohydrate availability for individual bacteria and their growth in the intestinal environment remains unclear. Here, we show that the availability of long-chain xylans (LCX), one of the most abundant dietary fibres in the human diet, promotes the growth of Bifidobacterium pseudocatenulatum in the adult human gut. Genomic and phenotypic analyses revealed that the availability of LCX-derived oligosaccharides is a fundamental feature of B. pseudocatenulatum, and that some but not all strains possessing the endo-1,4-ß-xylanase (BpXyn10A) gene grow on LCX by cleaving the xylose backbone. The BpXyn10A gene, likely acquired by horizontal transfer, was incorporated into the gene cluster for LCX-derived oligosaccharide utilisation. Co-culturing with xylanolytic Bacteroides spp. demonstrated that LCX-utilising strains are more competitive than LCX non-utilising strains even when LCX-derived oligosaccharides were supplied. In LCX-rich dietary interventions in adult humans, levels of endogenous B. pseudocatenulatum increased only when BpXyn10A was detected, indicating that LCX availability is a fitness determinant in the human gut. Our findings highlight the enhanced intestinal adaptability of bifidobacteria via polysaccharide utilisation, and provide a cornerstone for systematic manipulation of the intestinal microbiota through dietary intervention using key enzymes that degrade polysaccharide as biomarkers.

Influence of thermal stress and bleaching on heterotrophic feeding of two scleractinian corals on pico-nanoplankton.

The feeding strategies of Montipora digitata and Porites lutea, two dominant corals in the Okinawan reefs, were investigated. The focus was on pico- and nanoplankton feeding efficiencies, using 6-h incubations. Although healthy M. digitata consumed from 72% to 87% more pico-nanoplankton cells than P. lutea, feeding rates of bleached corals of both species were similarly low at heat stress (33 °C). Heterotrophic carbon acquisition with respect to dark respiration varied from 3% to 65% in M. digitata and from 7% to 68% in P. lutea. A decrease in the feeding efficiency of bleached M. digitata under heat stress shows its vulnerability to water heating events. Feeding rates of P. lutea were low under all conditions and treatments; therefore, this species is less vulnerable to heat stress due to the strategy of meeting metabolic costs by using translocated organic matter from endoliths and selecting pico-nanoplankton cells with a high C/N ratio.

Identification of Smad-dependent and -independent signaling with transforming growth factor-ß type 1/2 receptor inhibition in palatogenesis.

TGF-ß signaling is one of important function during palatal fusion. Three types of TGF-ß receptor (TßR1, TßR2, and TßR3) have been identified, and play essential roles in mechanisms leading to palatal fusion. However, the balance between Smad-dependent/-independent signaling during palatal fusion with inhibited TßR1/2 functions is not fully understood. The objective of this study was to investigate palatal fusion via TGF-ß signaling when TßR1 and TßR2, but not TßR3, were inhibited. In addition, the present study examined the functional balance between Smad-dependent/-independent signaling and related gene expression. Palatal organ cultures were treated with TßR1/2 inhibitor in vitro. Control palates were cultured without inhibitor. We observed histological phenotype of palatal fusion, and evaluation of expression pattern by Western blot or real time RT-PCR. Palatal organ cultures treated with the inhibitor did not fuse and the medial edge epithelium remained at embryonic 13 day +72 h in culture. The inhibitor decreased TßR1 and TßR2 expression by approximately 90%, but did not affect TßR3 expression. The expression of p-Smad2 and p-Smad3 was significantly decreased in treated palates compared with controls. The expression of p-Smad4 was slightly decreased in treated palates compared with controls. Smad-independent signaling was also affected by the inhibitor; p-ERK, p-JNK, and p-p38 expressions was significantly reduced in treated palates compared with controls. The expression of transcription factors (Runx1 and Msx1) and extracellular matrix proteins (MMP2/13) was also significantly decreased by inhibitor exposure. Treatment with TßR1/2 inhibitor altered the patterns of the Smad-dependent and -independent signaling pathways during palatal fusion.

Draft Genome Sequence of Streptomyces spongiicola Strain 531S, an Actinobacterium Isolated from Marine Sediment.

Streptomyces spongiicola strain 531S (NBRC 113560) was isolated from marine sediment on a beach on Sesoko Island (Okinawa, Japan). We report here the draft genome sequence of S. spongiicola 531S, in which 24 potential secondary metabolite gene clusters were predicted with antiSMASH.

Localization and density of Porphyromonas gingivalis and Tannerella forsythia in gingival and subgingival granulation tissues affected by chronic or aggressive periodontitis.

Porphyromonas gingivalis and Tannerella forsythia have been thought to be associated with periodontitis; however comprehensive histopathological localization of bacteria in affected human periodontal tissues is not well documented. In the present study, we examined formalin-fixed paraffin-embedded gingival and subgingival granulation tissues from 71 patients with chronic periodontitis and 11 patients with aggressive periodontitis, using immunohistochemistry with novel monoclonal antibodies specific to P. gingivalis or T. forsythia, together with quantitative real-time polymerase chain reaction for each bacterial DNA. Immunohistochemisty revealed both bacterial species extracellularly, as aggregates or within bacterial plaque, and intracellularly in stromal inflammatory cells, squamous epithelium, and capillary endothelium of granulation tissue. Combined analysis with the results from polymerase chain reaction suggested that localization and density of T. forsythia is closely associated with those of P. gingivalis, and that bacterial density is a factor responsible for the cell-invasiveness and tissue-invasiveness of these periodontal bacteria. Detection of these bacteria in the capillary endothelium in some samples suggested possible bacterial translocation into the systemic circulation from inflamed gingival and subgingival granulation tissues. Immunohistochemistry with the novel antibodies showed high specificity and sensitivity, and can be used to locate these periodontal bacteria in routinely-used formalin-fixed paraffin-embedded human tissue sections from systemic locations.

Propionibacterium acnes-derived insoluble immune complexes in sinus macrophages of lymph nodes affected by sarcoidosis.

BACKGROUND: Propionibacterium acnes is thought to be a causative agent of sarcoidosis. Patients with sarcoidosis have circulating immune complexes. We attempted to detect P. acnes-derived immune complexes in sarcoid lesions. METHODS: We evaluated formalin-fixed and paraffin-embedded lymph node samples from 38 sarcoidosis patients and 90 non-sarcoidosis patients (27 patients with necrotizing lymphadenitis, 28 patients with reactive lymphadenitis, 16 patients with colon cancer, 19 patients with gastric cancer) by immunohistochemistry using anti-human immunoglobulins (IgG, IgA, and IgM) and complement (C1q and C3c) antibodies, and a P. acnes-specific monoclonal antibody (PAB antibody) that reacts with the membrane-bound lipoteichoic acid of P. acnes. RESULTS: Small round bodies (SRBs) bound to IgA, IgM, or IgG were detected in sinus macrophages, in 32 (84%), 32 (84%), or 11 (29%) sarcoid samples, respectively, and in 19 (21%), 26 (29%), or no (0%) control samples, respectively. Some of these insoluble immune complexes (IICs) also bound to C1q and C3c. We developed a microwave treatment followed by brief trypsin digestion (MT treatment) to detect PAB-reactive SRBs bound to immunoglobulins (IIC-forming P. acnes). MT treatment revealed abundant IIC-forming P. acnes in most (89%) of the sarcoid samples and sparse distribution in some (20%) of the control samples with lymphadenitis, but no IIC-forming P. acnes was detected in control samples without inflammation. IIC-forming P. acnes were mostly bound to both IgA and IgM. The PAB-reactive antigen and immunoglobulins were both located at the peripheral rim of the IIC-forming P. acnes. Conventional electron microscopy identified many SRBs (0.5-2.0 µm diameter) in sinus macrophages of sarcoid lymph nodes with many IIC-forming P. acnes, some of which were in phagolysosomes with a degraded and lamellar appearance. CONCLUSIONS: P. acnes-derived IICs in sinus macrophages were frequent and abundant in sarcoid lymph nodes, suggesting a potential etiologic link between sarcoidosis and this commensal bacterium.

The evaluation of public health nursing students' skill achievement levels by educational improvement.

OBJECTIVE: This study aimed to evaluate the educational improvement outcomes after introducing undergraduate public health nursing electives from the perspective of students, teachers, and public health nurses. DESIGN AND SAMPLE: Post-test design with control group was used. We surveyed three groups of participants on the relevance of achieved skill items. These groups were as follows: fourth-year students, teaching staff from seven universities, and public health nurses from special districts of Tokyo who had pioneered the introduction of public health nursing electives. MEASURES: The three participant groups were surveyed in 2013 and 2014 to compare the students' levels of achievement in the 98 skill items presented by the Japanese Ministry of Health, Labour, and Welfare. Practical training introduced educational reform in 2014. Anonymous self-administrated questionnaires were used. RESULTS: The mean achievement rates of the 98 items in 2013 and 2014 were 67.9% and 72.6%, respectively, for students; 40.9% and 59.7% for teachers; and 44.4% and 59.4% for PHNs. A significant difference was observed between ratios of 2013 and 2014 in all three groups. CONCLUSIONS: The results demonstrated that the introduction of educational reform increased the achievement rates of students through the electives.

The different roles of innate immune receptors in inflammation and carcinogenesis between races.

Innate immune factors exert widespread effects on cytokine secretion, cell survival, autophagy, and apoptosis. Nucleotide-binding and oligomerization domain-like receptors (NLRs) are members of the innate immune system in the cytosol that sense pathogens, endogenous danger molecules such as uric acid, and pollutants. Nucleotide-binding oligomerization domain-containing protein 1 and 2 (NOD1 and NOD2) are components of NLR family, and ligands of these factors are γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP) and muramyl dipeptide (MDP), respectively. Upon recognition of ligands, NOD1 and NOD2 induce the production of inflammatory cytokines and transcription factors including interleukin-6 (IL-6) and nuclear factor-κB (NF-κB). We examined the function of NOD1 and NOD2 in innate immunity, with a focus on their differing roles in disease pathogenesis between Japanese and Caucasian populations. Susceptibility to several immune-related diseases, including Crohn's disease, colorectal and breast cancers, and graft-versus-host-disease (GVHD) showed a correlation with genetic variants of NOD2 in Caucasian, but not in Japanese, populations. This difference may be primarily due to the fact that three major NOD2 SNPs (R702W, G908R, L1007insC) prevalent in Caucasians are rare or absent in Japanese populations. Because NLR has diverse effects on immune function, it is possible that many as yet uncharacterized immune-related diseases will also show different susceptibilities between races due to the different ratio of genetic variants in innate immune genes.

Genome analysis of three novel lytic Vibrio coralliilyticus phages isolated from seawater, Okinawa, Japan.

Three novel Vibrio phages were isolated from seawater in Okinawa. The Vibrio phage RYC infected Vibrio coralliilyticus SWA 07, while Vibrio phages CKB-S1 and CKB-S2 infected the coral pathogen V. coralliilyticus P1 (LMG 23696). The Vibrio phages CKB-S1 and CKB-S2 displayed head-tail structures whereas the Vibrio phage RYC showed a tailless non-enveloped capsid. All these Vibrio phages contained linear and double-stranded DNA. The whole genome sequencing revealed that Vibrio phage RYC has a larger genome size compared to Vibrio phages CKB-S1 and CKB-S2, and six tRNAs genes were found only in Vibrio phage RYC. Genome-wide comparison showed that Vibrio phage CKB-S1 was closely related, but was not identical, to Vibrio parahaemolyticus phages VP16T and VP16C. Meanwhile, the Vibrio phages RYC and CKB-S2 did not show high genome-wide similarity to any phages. These results suggest that the Vibrio phages CKB-S1, CKB-S2 and RYC are novel phages, which need further exploration, especially for their potential applications in phage therapy.

Radiopathological Features and Identification of Mycobacterial Infections in Granulomatous Nodules Resected from the Lung.

BACKGROUND: Pulmonary granulomas are sometimes resected because they resemble lung cancer and false-positive findings come through from positron emission tomography (PET) using 18fluorine-fluorodeoxyglucose (18F-FDG). Mycobacterial infection is a common cause of granulomas. OBJECTIVE: The purpose of this study was to evaluate the radiopathological features and the methods for identifying mycobacterial infections in granulomatous nodules resected from the lung. METHODS: Thirty-five solitary lesions resected because of suspected lung cancer were enrolled, including 22 nonfungal granulomatous lesions and 13 benign lesions as controls. The radiological, microbiological, and histological findings were reviewed. To identify mycobacterial infection, immunohistochemical (IHC) staining, IS6110 polymerase chain reaction (PCR), and real-time PCR for the detection of Mycobacterium tuberculosis (TB) were performed using formalin-fixed and paraffin-embedded tissue specimens. The correlations between the radiopathological features and the median maximum standardized uptake value (SUVmax) of 18F-FDG PET were also evaluated. RESULTS: Mycobacteria were isolated from the cultures of 10 of the granulomatous lesions, including TB from 2 and Mycobacterium avium complex from 8. The mean size of the nodules in the culture-positive group was significantly larger than that of those in the culture-negative group (30.5 ± 13.1 vs. 15.1 ± 6.3 mm, p = 0.003). IHC stainings were positive in 15 granulomas. Eight granulomas were positive in IS6110 PCR, and 7 of them were also positive in real-time PCR. SUVmax was ≥2.5 in all of the PCR-positive granulomas. CONCLUSION: The most frequent cause of granulomatous lesions was mycobacterial infection. It seemed that the culture result was associated with nodule size and that the results of IS6110 were associated with 18F-FDG-uptake.

Frequency of Propionibacterium acnes Infection in Prostate Glands with Negative Biopsy Results Is an Independent Risk Factor for Prostate Cancer in Patients with Increased Serum PSA Titers.

BACKGROUND: Propionibacterium acnes has recently been implicated as a cause of chronic prostatitis and this commensal bacterium may be linked to prostate carcinogenesis. The occurrence of intracellular P. acnes infection in prostate glands and the higher frequency of P. acnes-positive glands in radical prostatectomy specimens from patients with prostate cancer (PCa) than in those from patients without PCa led us to examine whether the P. acnes-positive gland frequency can be used to assess the risk for PCa in patients whose first prostate biopsy, performed due to an increased prostate-specific antigen (PSA) titer, was negative. METHODS: We retrospectively collected the first and last prostate biopsy samples from 44 patients that were diagnosed PCa within 4 years after the first negative biopsy and from 36 control patients with no PCa found in repeated biopsy for at least 3 years after the first biopsy. We evaluated P. acnes-positive gland frequency and P. acnes-positive macrophage number using enzyme-immunohistochemistry with a P. acnes-specific monoclonal antibody (PAL antibody). RESULTS: The frequency of P. acnes-positive glands was higher in PCa samples than in control samples in both first biopsy samples and in combined first and last biopsy samples (P < 0.001). A frequency greater than the threshold (18.5 and 17.7, respectively) obtained by each receiver operating characteristic curve was an independent risk factor for PCa (P = 0.003 and 0.001, respectively) with odds ratios (14.8 and 13.9, respectively) higher than those of serum PSA titers of patients just before each biopsy (4.6 and 2.3, respectively). The number of P. acnes-positive macrophages did not differ significantly between PCa and control samples. CONCLUSIONS: These results suggested that the frequency of P. acnes-positive glands in the first negative prostate biopsy performed due to increased PSA titers can be supportive information for urologists in planning repeated biopsy or follow-up strategies.

13C and 15N assimilation and organic matter translocation by the endolithic community in the massive coral Porites lutea.

Corals evolved by establishing symbiotic relationships with various microorganisms (the zooxanthellae, filamentous algae, cyanobacteria, bacteria, archaea, fungi and viruses), forming the 'coral holobiont'. Among them, the endolithic community is the least studied. Its main function was considered to be translocation of photo-assimilates to the coral host, particularly during bleaching. Here, we hypothesize that (i) endolithic algae may show similar primary production rates in healthy or bleached corals by changing their pigment ratios, and therefore that similar production and translocation of organic matter may occur at both conditions and (ii) diazotrophs are components of the endolithic community; therefore, N2 fixation and translocation of organic nitrogen may occur. We tested these hypotheses in incubation of Porites lutea with 13C and 15N tracers to measure primary production and N2 fixation in coral tissues and endoliths. Assimilation of the 13C atom (%) was observed in healthy and bleached corals when the tracer was injected in the endolithic band, showing translocation in both conditions. N2 fixation was found in coral tissues and endolithic communities with translocation of organic nitrogen. Thus, the endolithic community plays an important role in supporting the C and N metabolism of the holobiont, which may be crucial under changing environmental conditions.

Cortical Actin Alteration at the Matrix-Side Cytoplasm in Lung Adenocarcinoma Cells and Its Significance in Invasion.

OBJECTIVES: Cortical actin is a thin layer of filamentous (F-)actin that lies beneath the plasma membrane, and its role in pathophysiology remains unclear. We investigated the subcellular localization of cortical actin by the histopathological and experimental studies of lung adenocarcinomas. MATERIALS AND METHODS: The subcellular localization of cortical actin was studied in surgically resected lung adenocarcinomas tissues and in 3-dimensionally cultured lung adenocarcinoma A549 cells. RESULTS: In normal type II alveolar cells and the bronchiolar epithelium, cortical actin was localized to the apical-side cytoplasm. In invasive adenocarcinoma cells, cortical actin was frequently localized to the matrix side. The degree of cortical actin localized to the matrix side was associated with the loss of basement membrane and a poor prognosis. In A549 cell spheroids cultured in a type I collagen and basement membrane extract Matrigel™ mixed gel, cortical F-actin was localized to the matrix side with phosphorylated myosin light chain. Super-resolution and electron microscopy results suggest that compact wrinkling of the plasma membrane by myosin-mediated F-actin contraction is an explanation for cortical actin accumulation at the matrix side. The myosin II inhibitor blebbistatin suppressed the 3-dimensional collective migration of A549 cells induced by constitutively active Cdc42 and MT1-MMP. CONCLUSION: Cortical actin accumulation at the matrix-side cytoplasm of cancer cells occurs in invasive lung adenocarcinomas and it possibly participates in the migration of cancer cells through myosin-mediated contraction.

Evaluation of skill achievement levels and practical experiences of public health nursing students before and after the introduction of the public health nursing course as an elective.

Objective To equip public health nurses (PHNs) with higher qualifications, PHN education is shifting from an integrated curriculum for PHNs and registered nurses to a specific elective system of undergraduate or postgraduate programs. Most colleges in the special wards of Tokyo introduced the elective system in 2014 before the remaining areas. The outcomes of this must be evaluated. This study aimed to evaluate the achievement levels and practical experiences of PHN students at seven colleges in the special wards before and after introduction of the PHN course as an elective.Method Self-administered, anonymous questionnaires were completed by senior PHN students at seven colleges in the special wards who underwent training in 2013, the last year of an integrated curriculum, and in 2014, the first year of the elective system. The target numbers of participants were 663 in 2013 and 136 in 2014 with 20 students from each school exposed to the elective system. Our study focused on whether they achieved the 98 "technical items of PHN training and achievement levels at the time of graduation" required by the Ministry of Health, Labour and Welfare. The study also determined whether participants obtained practical experience in 15 items developed by the special wards based on the standards set for training.Results In 2013, there were 348 total responses (52.5%) and 310 valid responses. In 2014, there were 136 total responses (88.2%) and 120 valid responses. The average achievement rate at which the student answered, "I was able to arrive at it," at an arrival degree level for the 98 technical items was 72.6% in 2014, an increase compared to the 67.9% obtained in 2013. Moreover, the average practical experience rate at which the student answered, "I was able to have an experience," regarding the 15 items was 85.7% in 2014, which constituted an increase compared to 70.5% attained in 2013. However, the number of items with an achievement rate of more than 80% remained at 31 (31.6%) in 2014.Conclusion Increasing percentages of average achievement and experience rate suggest that the lecture and training have improved, and highly motivated students could have been selected in the PHN program. However, students did not achieve the target rate for many of the items. Thus, there was a time limit to experience the technical item/the experience item.

Autophagy Induced by Intracellular Infection of Propionibacterium acnes.

BACKGROUND: Sarcoidosis is caused by Th1-type immune responses to unknown agents, and is linked to the infectious agent Propionibacterium acnes. Many strains of P. acnes isolated from sarcoid lesions cause intracellular infection and autophagy may contribute to the pathogenesis of sarcoidosis. We examined whether P. acnes induces autophagy. METHODS: Three cell lines from macrophages (Raw264.7), mesenchymal cells (MEF), and epithelial cells (HeLa) were infected by viable or heat-killed P. acnes (clinical isolate from sarcoid lymph node) at a multiplicity of infection (MOI) of 100 or 1000 for 1 h. Extracellular bacteria were killed by washing and culturing infected cells with antibiotics. Samples were examined by colony assay, electron-microscopy, and fluorescence-microscopy with anti-LC3 and anti-LAMP1 antibodies. Autophagy-deficient (Atg5-/-) MEF cells were also used. RESULTS: Small and large (≥5 µm in diameter) LC3-positive vacuoles containing few or many P. acnes cells (LC3-positive P. acnes) were frequently found in the three cell lines when infected by viable P. acnes at MOI 1000. LC3-positive large vacuoles were mostly LAMP1-positive. A few small LC3-positive/LAMP1-negative vacuoles were consistently observed in some infected cells for 24 h postinfection. The number of LC3-positive P. acnes was decreased at MOI 100 and completely abolished when heat-killed P. acnes was used. LC3-positive P. acnes was not found in autophagy-deficient Atg5-/- cells where the rate of infection was 25.3 and 17.6 times greater than that in wild-type Atg5+/+ cells at 48 h postinfection at MOI 100 and 1000, respectively. Electron-microscopic examination revealed bacterial cells surrounded mostly by a single-membrane including the large vacuoles and sometimes a double or multi-layered membrane, with occasional undigested bacterial cells in ruptured late endosomes or in the cytoplasm. CONCLUSION: Autophagy was induced by intracellular P. acnes infection and contributed to intracellular bacterial killing as an additional host defense mechanism to endocytosis or phagocytosis.

The northern limit of corals of the genus Acropora in temperate zones is determined by their resilience to cold bleaching.

The distribution of corals in Japan covers a wide range of latitudes, encompassing tropical to temperate zones. However, coral communities in temperate zones contain only a small subset of species. Among the parameters that determine the distribution of corals, temperature plays an important role. We tested the resilience to cold stress of three coral species belonging to the genus Acropora in incubation experiments. Acropora pruinosa, which is the northernmost of the three species, bleached at 13 °C, but recovered once temperatures were increased. The two other species, A. hyacinthus and A. solitaryensis, which has a more southerly range than A. pruinosa, died rapidly after bleaching at 13 °C. The physiological effects of cold bleaching on the corals included decreased rates of photosynthesis, respiration, and calcification, similar to the physiological effects observed with bleaching due to high temperature stress. Contrasting hot bleaching, no increases in antioxidant enzyme activities were observed, suggesting that reactive oxygen species play a less important role in bleaching under cold stress. These results confirmed the importance of resilience to cold stress in determining the distribution and northern limits of coral species, as cold events causing coral bleaching and high mortality occur regularly in temperate zones.