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The purpose of this study was to examine whether anti-transgender rights legislation among state legislators is associated with increased suicide- and depression-related Internet searches. Employing a quasi-experimental non-equivalent control group design, we focused on bills that were introduced to state legislatures from July 2019 to July 2020. As our panel is constructed of 51 states/territories over a 52-week time frame, our final dataset is composed of 2,652 observations. Results showed that states’ passing of anti-transgender rights bills were linked with suicide- and depression-related Internet searches. Second, introducing or debating the bills did not have an association with Internet searches. Third, the defeat of anti-transgender bills was linked with fewer depression-related searches. Finally, the LGBT context in the state affected the results: anti-transgender legislation had a particularly strong association with suicide-related Internet searches when the state had a high LGBT population density.

Although outpatient specialty palliative-care clinics improve outcomes, there is no consensus on who should be referred or the optimal timing for referral. In response to this issue, we did a Delphi study to develop consensus on a list of criteria for referral of patients with advanced cancer at secondary or tertiary care hospitals to outpatient palliative care. 60 international experts (26 from North America, 19 from Asia and Australia, and 11 from Europe) on palliative cancer care rated 39 needs-based criteria and 22 time-based criteria in three iterative rounds. Nearly all experts responded in each round. Consensus was defined by an a-priori agreement of 70% or more. Panellists reached consensus on 11 major criteria for referral: severe physical symptoms, severe emotional symptoms, request for hastened death, spiritual or existential crisis, assistance with decision making or care planning, patient request for referral, delirium, spinal cord compression, brain or leptomeningeal metastases, within 3 months of advanced cancer diagnosis for patients with median survival of 1 year or less, and progressive disease despite second-line therapy. Consensus was also reached on 36 minor criteria for specialist palliative-care referral. These criteria, if validated, could provide guidance for identification of patients suitable for outpatient specialty palliative care.

Tissue-clearing techniques are powerful tools for biological research and pathological diagnosis. Here, we describe advanced clear, unobstructed brain imaging cocktails and computational analysis (CUBIC) procedures that can be applied to biomedical research. This protocol enables preparation of high-transparency organs that retain fluorescent protein signals within 7–21 d by immersion in CUBIC reagents. A transparent mouse organ can then be imaged by a high-speed imaging system (>0.5 TB/h/color). In addition, to improve the understanding and simplify handling of the data, the positions of all detected cells in an organ (3–12 GB) can be extracted from a large image dataset (2.5–14 TB) within 3–12 h. As an example of how the protocol can be used, we counted the number of cells in an adult whole mouse brain and other distinct anatomical regions and determined the number of cells transduced with mCherry following whole-brain infection with adeno-associated virus (AAV)-PHP.eB. The improved throughput offered by this protocol allows analysis of numerous samples (e.g., >100 mouse brains per study), providing a platform for next-generation biomedical research. Transparent organs are obtained, with retained fluorescent protein signals, upon clearing by immersion in the appropriate CUBIC reagent. The positions of all the cells can be determined using the described software.

In a trial involving 1033 patients hospitalized with Covid-19, the addition of baricitinib to remdesivir was associated with shorter recovery time, particularly among patients receiving high-flow oxygen, and with a 30% higher odds of improvement at day 15 than remdesivir alone. Adverse events were less frequent with the combination therapy.

Sunlight exposure contributes to human health; however, excessive light exposure to skin, especially ultraviolet B (UV-B), can produce high amounts of reactive oxygen species (ROS) and induce inflammation. Some antioxidants, such as squalene, can prevent UV-B-induced inflammation. C34H58 botryococcene is the most common triterpene hydrocarbon produced by green alga Botryococcus braunii; it is biosynthesized via a pathway similar to squalene and appears to have a similar chemical structure to squalene. However, there are no reports on the bioactivity of botryococcene. In this study, we evaluated that botryococcene can prevent the skin photoaging. Using ESR assay, botryococcene could not scavenge any ROS. However, treatment of epidermis cells with the botryococcene significantly suppressed intracellular ROS production by hydrogen peroxide (H2O2) and attenuated H2O2 cytotoxicity. Botryococcene enhanced the antioxidant enzymes in gastric cells, thus botryococcene may scavenge ROS indirectly, not directly. Moreover, botryococcene inhibited production of intracellular interleukin-1 and exhibited suppression of melanogenesis activity by UV-B irradiation. Addition of botryococcene-treated epidermal cells culture medium mitigated the increase in matrix metalloproteinase-1 production and the decrease in type I collagen production induced by UV-B irradiation in dermis cells. These results showed that botryococcene has anti-photoaging effects, including preventing wrinkles and blemishes on the skin.

Microalgae have attracted significant attention worldwide as one of the most promising feedstock fossil fuel alternatives. However, there are a few challenges for algal fuels to compete with fossil fuels that need to be addressed. Therefore, this study reviews the R&D status of microalgae-based polyculture and biocrude oil production, along with wastewater treatment. Mixotrophic algae are free to some extent from light restrictions using organic matter and have the ability to grow well even in deep water-depth cultivation. It is proposed that integrating the mixotrophic microalgae polyculture and wastewater treatment process is the most promising and harmonizing means to simultaneously increase capacities of microalgae biomass production and wastewater treatment with a low land footprint and high robustness to perturbations. A large amount of mixotrophic algae biomass is harvested, concentrated, and dewatered by combining highly efficient sedimentation through flocculation and energy efficient filtration, which reduce the carbon footprint for algae fuel production and coincide with the subsequent hydrothermal liquefaction (HTL) conversion. HTL products are obtained with a relatively low carbon footprint and separated into biocrude oil, solid, aqueous, and gas fractions. Algae biomass feedstock-based HTL conversion has a high biocrude oil yield and quality available for existing oil refineries; it also has a bioavailability of the recycled nitrogen and phosphorus from the aqueous phase of algae community HTL. The HTL biocrude oil represents higher sustainability than conventional liquid fuels and other biofuels for the combination of greenhouse gas (GHG) and energy return on investment (EROI). Deep water-depth polyculture of mixotrophic microalgae using sewage has a high potential to produce sustainable biocrude oil within the land area of existing sewage treatment plants in Japan to fulfill imported crude oil.

Radiation induces various changes in biological specimens; however, the evaluation of these changes is usually complicated and can be achieved only through investment in time and labor. Optical methods reduce the cost of such evaluations as they require less pretreatment of the sample, are adaptable to high-throughput screening and are easy to automate. Optical methods are also advantageous, owing to their real-time and onsite evaluation capabilities. Here, we discuss three optical technologies to evaluate the effects of radiation on biological samples: single-molecule tracking microscopy to evaluate the changes in the physical properties of DNA, Raman spectral microscopy for dosimetry using human hair and second-harmonic generation microscopy to evaluate the effect of radiation on the differentiation of stem cells. These technologies can also be combined for more detailed information and are applicable to other biological samples. Although optical methods are not commonly used to evaluate the effects of radiation, advances in this technology may facilitate the easy and rapid assessment of radiation effects on biological samples.

System level understanding of the cell requires detailed description of the cell state, which is often characterized by the expression levels of proteins. However, understanding the cell state requires comprehensive information of the cell, which is usually obtained from a large number of cells and their disruption. In this study, we used Raman spectroscopy, which can report changes in the cell state without introducing any label, as a non-invasive method with single cell capability. Significant differences in Raman spectra were observed at the levels of both the cytosol and nucleus in different cell-lines from mouse, indicating that Raman spectra reflect differences in the cell state. Difference in cell state was observed before and after the induction of differentiation in neuroblastoma and adipocytes, showing that Raman spectra can detect subtle changes in the cell state. Cell state transitions during embryonic stem cell (ESC) differentiation were visualized when Raman spectroscopy was coupled with principal component analysis (PCA), which showed gradual transition in the cell states during differentiation. Detailed analysis showed that the diversity between cells are large in undifferentiated ESC and in mesenchymal stem cells compared with terminally differentiated cells, implying that the cell state in stem cells stochastically fluctuates during the self-renewal process. The present study strongly indicates that Raman spectral morphology, in combination with PCA, can be used to establish cells' fingerprints, which can be useful for distinguishing and identifying different cellular states.

Fluorescence live imaging has become an essential methodology in modern cell biology. However, fluorescence requires excitation light, which can sometimes cause potential problems, such as autofluorescence, phototoxicity, and photobleaching. Furthermore, combined with recent optogenetic tools, the light illumination can trigger their unintended activation. Because luminescence imaging does not require excitation light, it is a good candidate as an alternative imaging modality to circumvent these problems. The application of luminescence imaging, however, has been limited by the two drawbacks of existing luminescent protein probes, such as luciferases: namely, low brightness and poor color variants. Here, we report the development of bright cyan and orange luminescent proteins by extending our previous development of the bright yellowish-green luminescent protein Nano-lantern. The color change and the enhancement of brightness were both achieved by bioluminescence resonance energy transfer (BRET) from enhancedRenillaluciferase to a fluorescent protein. The brightness of these cyan and orange Nano-lanterns was ∼20 times brighter than wildtypeRenillaluciferase, which allowed us to perform multicolor live imaging of intracellular submicron structures. The rapid dynamics of endosomes and peroxisomes were visualized at around 1-s temporal resolution, and the slow dynamics of focal adhesions were continuously imaged for longer than a few hours without photobleaching or photodamage. In addition, we extended the application of these multicolor Nano-lanterns to simultaneous monitoring of multiple gene expression or Ca²⁺ dynamics in different cellular compartments in a single cell.