Efficient virus-induced gene silencing in apple, pear and Japanese pear using Apple latent spherical virus vectors

<p>Abstract</p> <p>Background</p> <p>Virus-induced gene silencing (VIGS) is an effective technology for the analysis of gene functions in plants. Though there are many reports on virus vectors for VIGS in plants, no VIGS vectors available for <it>Rosaceae </it>fruit trees were reported so far. We present an effective VIGS system in apple, pear, and Japanese pear using <it>Apple latent spherical virus </it>(ALSV) vectors.</p> <p>Results</p> <p>Inoculation of ALSV vectors carrying a partial sequence of endogenous genes from apple [ribulose-1, 5-bisphosphate carboxylase small subunit (<it>rbcS</it>), alpha subunit of chloroplast chaperonin (<it>CPN60a</it>), elongation factor 1 alpha (<it>EF-1a</it>), or actin] to the cotyledons of seeds by a particle bombardment induced highly uniform knock-down phenotypes of each gene on the true leaves of seedlings from 2~3 weeks after inoculation. These silencing phenotypes continued for several months. Northern blot and RT-PCR analyses of leaves infected with ALSV containing a fragment of <it>rbcS </it>gene showed that the levels of <it>rbcS</it>-mRNA drastically decreased in the infected apple and pear leaves, and, in reverse, <it>rbcS-</it>siRNAs were generated in the infected leaves. In addition, some of apple seedlings inoculated with ALSV vector carrying a partial sequence of a <it>TERMINAL FLOWER 1 </it>gene of apple (<it>MdTFL1</it>) showed precocious flowering which is expected as a knock-down phenotype of the silencing of <it>MdTFL1 </it>gene.</p> <p>Conclusions</p> <p>The ALSV-based VIGS system developed have provides a valuable new addition to the tool box for functional genomics in apple, pear, and Japanese pear.</p

Identifying bereaved grievers with greatest medical or social service needs in Japan

[Objective] Severe grief adversely affects the health of bereaved families, potentially burdening medical and community health services. Interventions for effective community health maintenance must identify the people likely to face severe effects of bereavement. The present study identified characteristics of mourners who experience severe grief within a year of bereavement to confirm whether this grief increased their reliance on Japanese medical and social services. [Design] We conducted a nationwide postal survey of Japanese bereaved within the previous year, to compare those reporting daily or overwhelming ‘heavy’ grief to those with less heavy grief, in terms of demographic and socioeconomic details, daily work and non-work activity, frequency of medical and social service use. [Setting/participants] In 2019, with the support of the Ministry of Education and the All Japan Funeral Co-Operation, we distributed approximately 5500 questionnaires to Japanese who had presided at funerals within the past year for anonymous return. By January of 2020, we received 1078 complete voluntary responses from bereaved Japanese. [Results] Half of the ‘heavy grief’ group (n=143) reported adverse effects on health and daily life, including needs for pharmacological, medical or welfare support. Losses of husbands or children were particularly connected to severe grief; ‘unexpected’ death from cancer caused the greatest shock. Employment (even part-time) buffered against severe grief; grief was greater for the unemployed and substantially worse for those who lost significant income at the same time as they lost loved ones. [Conclusion] These findings suggest that prior counselling should reduce the shock of bereavement and economic loss, which increases subsequent medical dependence. Medical professionals and community health workers can use the above factors to target in advance the family members in greatest danger of heavy grief, to intervene lest grief adversely affect their physical and psychological health after bereavement

Neuroprotective Effects of Lutein in the Retina

Although a large variety of pharmaceutical therapies for treating disease have been developed in recent years, there has been little progress in disease prevention. In particular, the protection of neural tissue is essential, because it is hardly regenerated. The use of nutraceuticals for maintaining the health has been supported by several clinical studies, including cross-sectional and interventional studies for age-related macular disease. However, mechanistic evidence for their effects at the molecular level has been very limited. In this review, we focus on lutein, which is a xanthophyll type of carotenoid. Lutein is not synthesized in mammals, and must be obtained from the diet. It is delivered to the retina, and in humans, it is concentrated in the macula. Here, we describe the neuroprotective effects of lutein and their underlying molecular mechanisms in animal models of vision-threatening diseases, such as innate retinal inflammation, diabetic retinopathy, and light-induced retinal degeneration. In lutein-treated mouse ocular disease models, oxidative stress in the retina is reduced, and its downstream pathological signals are inhibited. Furthermore, degradation of the functional proteins, rhodopsin (a visual substance) and synaptophysin (a synaptic vesicle protein also influenced in other neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease), the depletion of brain-derived neurotrophic factor (BDNF), and DNA damage are prevented by lutein, which preserves visual function. We discuss the possibility of using lutein, an antioxidant, as a neuroprotective treatment for humans

Unexplored Costs of Bereavement Grief in Japan: Patterns of Increased Use of Medical, Pharmaceutical, and Financial Services

Our Japan-wide survey (n=1, 078) revealed that bereaved who showed increased reliance on medical, pharmaceutical, and financial/legal services was in the ‘50’s age bracket, and for unemployed widows; it corresponded less with low annual income than with high income. Increased users reported their decline in physical health seriously influencing their work and lives, suggesting “presenteeism”. Moreover, they spent 2.7 times more for medical and pharmaceutical services than those reporting continual use, portending 4 to 10 times more Japanese government expense for this group, half of whom considered their own out-of-pocket expenses a financial burden