Used theme word : deer(main characters), soulmate (Bamboo and Bambi), traffic (the turtle), Falling (falling to under the sea) Goal : I want to show unexpected but structured stories to players like Quentin Tarantino's movie.

This is Mt. Vesuvius in Italy. One day, Mount Vesuvius, who thought it was a dormant volcano, suddenly began to explode. The mountain was accompanied by a huge earthquake and volcanic ash. Many of the animals and plants in the mountain were blown away by the explosion. In particular, there were many deer on Mount Vesuvius, many of which fell from the sky. Among the drops were a mix of leaves and shoes as well as deer. At the time of the eruption, a woodcutter remained unable to escape the eruption at Vesuvius. He had to avoid the rain of deer falling from the sky.. When the game starts, you can see the setting of moon night. In here, press the 'r' key on the initial page. The background music of the early pages begins after a brief rainstorm. The sound of the rain was put in because our game name is Rainy Deer. When the game starts, the time of survival and the number of lives remaining are marked on the left end. If you get hit by a deer, the number of lives decreases. Deer drops more and more quickly over time goes by. The goal of the game is to survive the falling deer.There are three items that help you survive. Heart increases the number of lives. Shoes increases the speed of the player. Leaves makes slow down the deer. Three items fall at once at the start of the game. You have to carefully choose what to eat as an initial item. After that, each item falls into a random position on a different time basis. Player should be careful not to incur the deer while eating the item. When the game is over, you can see the time you recorded. If you press the 'r' key once more on the ending page, you can play it again. When designing the game, I was inspired by the "Dung-Boo Game," which I often played as a child. Games like this are a form of game that you can often see when web games developed in the past. Nevertheless, what's unique about our games is that they can create dynamic cases, not just fall. The deer's speed is getting faster. Deer grows gradually every second. The amount falling also increases. Deer can control the amount of drop as an item. Games like this used to be one life, but we provide players with many lives. They can also fill their lives through items. The speed of the player can also be adjusted as an item. By adding multiple items to the existing game format, people often die trying to eat items. This situation increases the fear of avoiding deer falling from the top. This type of game usually uses a lot of cute images, where deer fall and bleed. This effect has increased the tension of the game. It also caused the blood to remain so, requiring more concentration to avoid the deer that continued to fall. When people played an alpha version of the game, the feedback we received most was that players didn't know whether they were hit by a deer or not. To solve this problem, we added a sound that matches an object that fits when the player is hit by a deer. TA got feedback asking us to add background music. I picked two pieces of music that fit our situation from my usual favorite game, Maple Story background music, and added music that my team mate liked to set up as starting music, game-going music and ending music. TA also advised that the deer's blood should not be removed quickly, but left to remain, giving it a grotteptic look. We deeply sympathized with this feedback and adjusted the options so that the deer remained in the dead position. I made a game for the first time in my life. By making this game, I realized how complicated the game I play is to make. Thinking that every move of the player was coded, I realized why the game company needed so many developers. As I thought that the game company made all the resources necessary for the game, designers and musicians belonging to the game production company respected me. Especially, for music, it is not easy to be inspired and you have to create different music from map to map. The music also has to go well with the artistic part of the game. Therefore, game companies should make efforts outside of game production, such as having to adjust each department's schedule well. Recognizing this process, I thought that all the resources that I could see in the game were made for this game, and I thought it was not a game I used to play. Through this project, I can enjoy games more. Imagery sources: deer : https://www.istockphoto.com/illustrations/clip-art-of-christmas-pixel-art?assettype=image&sort=mostpopular&mediatype=illustration&family=creative&phrase=clip%20art%20of%20christmas%20pixel%20art extra_life : https://tchol.org/explore/transparent-pixel.html blood : https://opengameart.org/content/bloodsplatter-and-bloodsplash-animation ground : https://www.reddit.com/r/PixelArt/comments/8ql7ol/ocnewbieccwip_ground_tiles_ive_been_working_on/ hit : https://www.reddit.com/r/PixelArt/comments/8ql7ol/ocnewbieccwip_ground_tiles_ive_been_working_on/ player : http://pixeljoint.com/forum/forum_posts.asp?TID=19431 slow_down : http://pixelartmaker.com/art/a13c31b62d93882 speed_up : http://pixelartmaker.com/art/63862e153250ee3 TiledBackground3 : https://arludus.itch.io/pixel-art-backgrounds-10-sky-cloud TiledBackground2 : https://busy.org/@dksart/alien-planet-pixel-art-weekly-contest-entry TiledBackground: https://www.deviantart.com/lord-lorzd/favourites/78826071/Insp Music sources : initial BGM : Maple story "forest of trials" https://www.youtube.com/watch?v=i5cQirTYXQk During game BGM : Maple story "Cunning City" https://www.youtube.com/watch?v=ryZJGm1XaIA Ending BGM : Dead silence https://www.youtube.com/watch?v=Wfp2KkDioi8 Item obtaining sound : The sound of the overwatch occupation Heating sound : https://effectdown.tistory.com/1 My team member : Sihwan Park

Varying pH of luminal fluid along the female reproductive tract is a physiological cue that modulates sperm motility. CatSper is a sperm-specific, pH-sensitive calcium channel essential for hyperactivated motility and male fertility. Multi-subunit CatSper channel complexes organize linear Ca²⁺ signaling nanodomains along the sperm tail. Here, we identify EF-hand calcium-binding domain-containing protein 9 (EFCAB9) as a bifunctional, cytoplasmic machine modulating the channel activity and the domain organization of CatSper. Knockout mice studies demonstrate that EFCAB9, in complex with the CatSper subunit, CATSPERζ, is essential for pH-dependent and Ca²⁺-sensitive activation of the CatSper channel. In the absence of EFCAB9, sperm motility and fertility is compromised, and the linear arrangement of the Ca²⁺ signaling domains is disrupted. EFCAB9 interacts directly with CATSPERζ in a Ca²⁺-dependent manner and dissociates at elevated pH. These observations suggest that EFCAB9 is a long-sought, intracellular, pH-dependent Ca²⁺ sensor that triggers changes in sperm motility.

A 45-yr-old female patient was admitted with one-month history of headache and progressive left hemiparesis. Brain magnetic resonance imaging (MRI) demonstrated a mass lesion in her right frontal lobe. Her brain tumor was confirmed as a small cell glioblastoma. Her follow-up brain MRI, taken at 8 months after her initial surgery demonstrated tumor recurrence in the right frontal lobe. Contrast-enhanced 7.0T brain magnetic resonance imaging (MRI) was safely performed before surgery and at the time of recurrence. Compared with 1.5T and 3.0T brain MRI, 7.0T MRI showed sharpened images of the brain tumor contexture with detailed anatomical information. The fused images of 7.0T and 1.5T brain MRI taken at the time of recurrence demonstrated no significant discrepancy in the positions of the anterior and the posterior commissures. It is suggested that 7.0T MRI can be safely utilized for better images of the maligant gliomas before and after surgery.

We report fast, non-scanning, wide-field two-photon fluorescence excitation with spectral and lifetime detection for in vivo biomedical applications. We determined the optical characteristics of the technique, developed a Gaussian flat-field correction method to reduce artifacts resulting from non-uniform excitation such that contrast is enhanced, and showed that it can be used for ex vivo and in vivo cellular-level imaging. Two applications were demonstrated: (i) ex vivo measurements of beta-amyloid plaques in retinas of transgenic mice, and (ii) in vivo imaging of sulfonated gallium(III) corroles injected into tumors. We demonstrate that wide-field two photon fluorescence excitation with flat-field correction provides more penetration depth as well as better contrast and axial resolution than the corresponding one-photon wide field excitation for the same dye. Importantly, when this technique is used together with spectral and fluorescence lifetime detection modules, it offers improved discrimination between fluorescence from molecules of interest and autofluorescence, with higher sensitivity and specificity for in vivo applications.

Key message

The non-intrinsic ABC proteins ABCI20 and ABCI21 are induced by light under HY5 regulation, localize to the ER, and ameliorate cytokinin-driven growth inhibition in young Arabidopsis thaliana seedlings. The plant ATP-binding cassette (ABC) I subfamily (ABCIs) comprises heterogeneous proteins containing any of the domains found in other ABC proteins. Some ABCIs are known to function in basic metabolism and stress responses, but many remain functionally uncharacterized. ABCI19, ABCI20, and ABCI21 of Arabidopsis thaliana cluster together in a phylogenetic tree, and are suggested to be targets of the transcription factor ELONGATED HYPOCOTYL 5 (HY5). Here, we reveal that these three ABCIs are involved in modulating cytokinin responses during early seedling development. The ABCI19, ABCI20 and ABCI21 promoters harbor HY5-binding motifs, and ABCI20 and ABCI21 expression was induced by light in a HY5-dependent manner. abci19 abci20 abci21 triple and abci20 abci21 double knockout mutants were hypersensitive to cytokinin in seedling growth retardation assays, but did not show phenotypic differences from the wild type in either control medium or auxin-, ABA-, GA-, ACC- or BR-containing media. ABCI19, ABCI20, and ABCI21 were expressed in young seedlings and the three proteins interacted with each other, forming a large protein complex at the endoplasmic reticulum (ER) membrane. These results suggest that ABCI19, ABCI20, and ABCI21 fine-tune the cytokinin response at the ER under the control of HY5 at the young seedling stage.

The epithelial cells of the gut form a physical barrier against the luminal contents. The collapse of this barrier causes inflammation, and its therapeutic restoration can protect the gut against inflammation. EGF enhances mucosal barrier function and increases colonocyte proliferation, thereby ameliorating inflammatory responses in the gut. Based on our previous finding that the insect peptide CopA3 promotes neuronal growth, we herein tested whether CopA3 could increase the cell proliferation of colonocytes, enhance mucosal barrier function, and ameliorate gut inflammation. Our results revealed that CopA3 significantly increased epithelial cell proliferation in mouse colonic crypts and also enhanced colonic epithelial barrier function. Moreover, CopA3 treatment ameliorated Clostridium difficile toxin As-induced inflammation responses in the mouse small intestine (acute enteritis) and completely blocked inflammatory responses and subsequent lethality in the dextran sulfate sodium-induced mouse model of chronic colitis. The marked CopA3-induced increase of colonocyte proliferation was found to require rapid protein degradation of p21(Cip1/Waf1), and an in vitro ubiquitination assay revealed that CopA3 directly facilitated ubiquitin ligase activity against p21(Cip1/Waf1). Taken together, our findings indicate that the insect peptide CopA3 prevents gut inflammation by increasing epithelial cell proliferation and mucosal barrier function.

We report that the Gm7068 (CatSpere) and Tex40 (CatSperz) genes encode novel subunits of a 9-subunit CatSper ion channel complex. Targeted disruption of CatSperz reduces CatSper current and sperm rheotactic efficiency in mice, resulting in severe male subfertility. Normally distributed in linear quadrilateral nanodomains along the flagellum, the complex lacking CatSperζ is disrupted at ~0.8 μm intervals along the flagellum. This disruption renders the proximal flagellum inflexible and alters the 3D flagellar envelope, thus preventing sperm from reorienting against fluid flow in vitro and efficiently migrating in vivo. Ejaculated CatSperz-null sperm cells retrieved from the mated female uterus partially rescue in vitro fertilization (IVF) that failed with epididymal spermatozoa alone. Human CatSperε is quadrilaterally arranged along the flagella, similar to the CatSper complex in mouse sperm. We speculate that the newly identified CatSperζ subunit is a late evolutionary adaptation to maximize fertilization inside the mammalian female reproductive tract.

RNA interference represents a potent intervention for cancer treatment but requires a robust delivery agent for transporting gene-modulating molecules, such as small interfering RNAs (siRNAs). Although numerous molecular approaches for siRNA delivery are adequate in vitro, delivery to therapeutic targets in vivo is limited by payload integrity, cell targeting, efficient cell uptake, and membrane penetration. We constructed nonviral biomaterials to transport small nucleic acids to cell targets, including tumor cells, on the basis of the self-assembling and cell-penetrating activities of the adenovirus capsid penton base. Our recombinant penton base chimera contains polypeptide domains designed for noncovalent assembly with anionic molecules and tumor homing. Here, structural modeling, molecular dynamics simulations, and functional assays suggest that it forms pentameric units resembling viral capsomeres that assemble into larger capsid-like structures when combined with siRNA cargo. Pentamerization forms a barrel lined with charged residues mediating pH-responsive dissociation and exposing masked domains, providing insight on the endosomolytic mechanism. The therapeutic impact was examined on tumors expressing high levels of HER3/ErbB3 that are resistant to clinical inhibitors. Our findings suggest that our construct may utilize ligand mimicry to avoid host attack and target the siRNA to HER3+ tumors by forming multivalent capsid-like structures.

This study presents a new multimodal imaging approach that includes high-frequency ultrasound, fluorescence intensity, confocal, and spectral imaging to improve the preclinical evaluation of new therapeutics in vivo. Here we use this approach to assess in vivo the therapeutic efficacy of the novel chemotherapy construct, HerDox during and after treatment. HerDox is comprised of doxorubicin non-covalently assembled in a viral-like particle targeted to HER2+ tumor cells, causing tumor cell death at over 10-fold lower dose compared to the untargeted drug, while sparing the heart. Whereas our initial proof-of-principle studies on HerDox used tumor growth/shrinkage rates as a measure of therapeutic efficacy, here we show that multimodal imaging deployed during and after treatment can supplement traditional modes of tumor monitoring to further characterize the particle in tissues of treated mice. Specifically, we show here that tumor cell apoptosis elicited by HerDox can be monitored in vivo during treatment using high frequency ultrasound imaging, while in situ confocal imaging of excised tumors shows that HerDox indeed penetrated tumor tissue and can be detected at the subcellular level, including in the nucleus, via Dox fluorescence. In addition, ratiometric spectral imaging of the same tumor tissue enables quantitative discrimination of HerDox fluorescence from autofluorescence in situ. In contrast to standard approaches of preclinical assessment, this new method provides multiple/complementary information that may shorten the time required for initial evaluation of in vivo efficacy, thus potentially reducing the time and cost for translating new drug molecules into the clinic.