生物化學

The emergence of antibacterial resistance has necessitated the development of alternative treatments, such as antimicrobial peptides (AMPs). AMPs are part of the innate immune systems of various organisms such as Momordica charantia L., a known medicinal plant in Southeast Asia. In this study, potential novel AMPs from M. charantia were derived in silico to provide prospective antibiotic alternatives using promising plant-based peptides. M. charantia protein sequences that were 500 amino acids long were digested using proteolytic enzymes, resulting in 3,621 peptides. Each resulting sequence was characterized as either AMP or Non-AMP using four statistical analysis tools, and those identified as AMPs were analyzed. This led to 102 AMPs, 53 of which were unregistered on the Data Repository for Antimicrobial Peptides, indicating that they have yet to be derived from other species. Six of the eight studied physicochemical properties show strong correlations with each other, suggesting that subsequent AMP design studies may focus on these six properties. As such, M. charantia may be a rich source of potential AMPs and, thereby, alternative antibiotics. The in vitro examination of these novel AMPs is also recommended to further understand their potential as alternative antibiotics sourced from locally available plants.

非血基質鐵/α-酮戊二酸依賴型雙氧化酶為巨大酵素家族，其在生物體內進行許多重要反應。我們選用小巢狀麴菌之 AsqJ 蛋白，其與 Fe2+ 及 α-酮戊二酸能催化 cyclopeptin 的去飽和、環氧化，形成 cyclopenin。根據 AsqJ 蛋白結構，我們將與受質結合的 9 個胺基酸位置以不同鹼基對隨機取代，以轉譯不同胺基酸，構築酵素活性庫，送入大腸桿菌篩選並觀察 AsqJ 突變蛋白分解非典型受質之表現。 我們成功篩選出能分解 X-Gal 之 AsqJ 突變株，代表我們能以改變 AsqJ 特定位置胺基酸，進而改變其活性位點，並分解與原先截然不同的受質，故此篩選平台深具生化技術應用潛力。我們並進一步將上述突變株進行純化、晶體培養及結構解析，以釐清反應機制，並與奈米抗體結合以增強活性。目前正嘗試分解環境汙染物結構相似物，期望對環境做出貢獻。

TDP-43是漸凍症(ALS)以及額顳葉失智症(FTLD)等神經退化性疾病的致病蛋白，在患者腦內會有不正常的類澱粉蛋白TDP-43錯誤折疊而產生蛋白質堆積，而不同堆積階段依序為單體(monomer)、多倍體(oligomer)、纖維(fiber)。其中纖維為許多神經退行性疾病的病理標誌，且相較於已有的 TDP-43 oligomer 單株抗體(Yu-Sheng Fang et al.(2014))，目前沒有對於 TDP-43 纖維抗體的研究，因此本實驗利用單株抗體技術所製造出對應TDP-43纖維的不同抗體進行純化及濃縮，找出對TDP-43 fiber專一性最高、結合效率及結合率皆最好抗體，以作為能夠檢測漸凍症患者血液中TDP-43 fiber的生物標誌(Biomarker)。 同時為了增加偵測的靈敏度，本實驗成功創新利用PMCA ( protein misfolding cyclic amplification) 技術快速放大纖維的數量以提高檢測準確度，搭配高專一性、高結合效率的編號3-2自製抗體作為Biomarker，只需五個小時便可以將極少量纖維的訊號約提高至原先的17倍，具有極高的靈敏性可以準確辨認出有無TDP-43纖維的樣本差別。

本研究針對臺灣恆春半島海域所採集的櫟葉指形軟珊瑚Sinularla querciformis進行天然物化學成分研究，由此珊瑚中分離出兩個新型菸草烷類型天然化合物，分別是Querciformolide G (1)與Querciformolide H (2)，以及兩個已知菸草烷類型天然化合物，分別是Sinulaparvalide B (3)與3,4:8,11-Bisepoxy-7-hydroxycembra-15(17)-en-1,12-olide (4)。上述化合物的物理性質和化學結構皆是由核磁共振儀、紅外線光譜儀和質譜儀等數據，以及比對相關化合物的文獻來分析確認。 針對化合物1-4進行抗發炎測試，對超氧陰離子產生和人中性粒細胞彈性蛋白酶釋放的抑製作用，發現化合物2針對彈性蛋白酶，有顯著的抑制效果。

神經細胞成熟的過程中可分成數個階段，每個階段間的轉換都伴隨著蛋白質的種類，RNA異構體、細胞結構與功能等全面性的轉變。但控制神經細胞在確切的時序下成熟的分子機制尚待研究。本研究發現 Paxillin 的新功能：當腦神經細胞在活體外培養至第七天時， Paxillin 的位點 Serine119 會被磷酸化 (p-PaxillinS119)，並從細胞質轉位進入至細胞核。我們使用 N2a 細胞以神經分化的模式來探討 p-PaxillinS119 進核的分子機制與功能，發現 p-PaxillinS119 進入細胞核需要位點 Serine119 被磷酸化，且分析後確認 Paxillin 的 LIM 結構域中帶有 PY-NLS 序列，分別為 P516/Y517 及 P575/Y576。我們發現 Paxillin 藉由轉運蛋白 Importin β2 辨識其 PY-NLS序列，進行蛋白間的交互作用後進入細胞核中。從螢光影像的分析，我們觀察到神經細胞的 p-PaxillinS119 在細胞核中會呈現顆粒狀，並與 RNA 剪接因子 P-SR 共定位在核斑點上。經由免疫共沉澱與細胞轉染的方式，我們證實位點 Serine119 突變，會影響 Paxillin 與 RNA 剪接因子的交互作用，及降低細胞分化與 RNA 剪接的程度。

在幹細胞治療中，並無法去偵測打入之幹細胞在人體內的位置及存在狀態。本研究創新在於(1)藉由使用新型奈米氧化鐵(IOP, iron oxide nanoparticles)來標定間葉幹細胞(MSCs, mesenchymal stem cells)，並能順利在磁振造影下成像 (2)同時確認IOP其安全性及效能。在實驗中先以X光繞射確認IOP粒徑平均10.4 nm，磁滯曲線證實不會產生磁性，CCK-8細胞測試中證實高濃度IOP(400 μg/mL)對間葉幹細胞存活影響小於17%，接著以普魯士藍染色觀察到IOP濃度愈高被幹細胞吸收愈多，平均每個細胞攝入2.79顆粒增加至3.47顆粒，並同時在磁振造影成像證實。最後高濃度IOP (400 μg/mL)會造成PGE2顯著性的下降55%，證實IOP可影響免疫調節但其角色仍待進一步研究。

原始生命的形成被推論是由化學分子透過自發性化學反應組成簡單的代謝路徑，再由這些路徑組合成為複雜的代謝網絡，最終形成原始的生命。由於自營性的代謝路徑可將無機性的CO₂固定為有機物可謂形成生命的前提，因此推論在自營性微生物所擁有的一種生物固碳路徑，可將CO₂固定進入三羧酸循環的「逆向三羧酸循環」被認為是原始細胞最初形成的代謝路徑之一。然而，以化學反應的自由能考量，自2-Oxoglutarate到Isocitrate 此固碳反應並非自發性反應，並由異檸檬酸脫氫酶催化。異檸檬酸脫氫酶在逆三羧酸循環中是決定固碳反應速率的關鍵酵素之一，在當今正向三羧酸循環中亦具重要的調節細胞能量代謝的功能。有鑑於此，本研究探討源自古老地球環境的嗜熱自營菌Aquifex aeolicus 中異檸檬酸脫氫酶的酵素性質，並期待本研究成果對生命起源的探究及發展新穎固碳技術能有所助益。

Previous research observed increased TNTs formation between hypoxic and normoxic neuroblastoma cells, aiding hypoxic cell survival. CHGA was identified as a potential factor in this process. This study compared CHGA expression and whether CHGA exists in TNTs in five cell lines, with SH-SY5Y showing the highest levels, followed by SK-N-BE(2)C, while the other three showed lower expression. Future studies will focus on the impact of CHGA on cell survival and its mechanisms.

In radiation treatments and manned interplanetary space travel, radiation is one of the biggest problems. The radiotolerance of cancer cells makes it necessary to apply high doses to surrounding healthy tissues by subjecting the cells to heavy stress. With regard to space travel (which involves a minimum travel time of 6 months) (1,2) the danger concerns cosmic radiation which is capable of inducing genetic mutations that, in turn, can evolve into very serious pathologies, such as cancer, damage to dendrites consequently compromising synapses. The project is aimed at developing a technology that can address these issues and aims to make human DNA radioresistant. This study involves a nucleosome-binding protein called DSUP (Damage Suppressor Protein) unique to the tardigrade Ramazzottius Varieornatus and the subject of its radioresistance. It can theoretically safeguard genetic material damaged by radiation. *Internship theme at the Pino Torinese Astronomical Observatory and the DISIT-UPO Environmental Molecular Toxicology Laboratory. The study molecule: Numerous tests have been carried out through the "in silico" approach geared toward mathematical modeling of its protein structure and complex mechanisms of action simulated through artificial intelligence systems, followed by direct laboratory analysis involving biosynthesis of DSUP by genetically modified bacteria and related tests. Both approaches, applied synergistically, aim to make it accessible and useful for the protection of human health.

Most people in the country tend to consume soda as part of their daily lives, without thinking about the health consequences that this entails. And the fact is that 墨西哥 is considered the main consumer of soda in the world (Universidad Nacional Autónoma de México [UNAM], 2019), and not only that, it also tops the lists of obesity and diabetes (Procuraduría Federal del Consumidor [PROFECO], 2018). A bad diet is one of the main causes of these medical conditions, so it is necessary to rethink the foods of daily consumption. In this project, the development of a carbonated beverage with three main ingredients: purple corn, hibiscus, and pineapple, which contain diverse nutritional properties in addition to having significant amounts of antioxidants (Secretaría de Cultura,2020; Sumaya et al., 2014; Kongsuwan et al., 2009), is established as an objective. In addition, antioxidants act as regulators of cellular aging and prevent chronic diseases (Vilaplana, 2007). For this reason, the elaboration of a beverage with these characteristics is considered harmless and a good option for people with a chronic disease or who are prone to it, as well as a healthy alternative to the consumption of soft drinks. In this project, an experimental methodology was followed to prepare the nutritional beverage, which had nutritional properties, such as vitamin and antioxidant content, in addition to having a pleasant taste and texture.