生物化學

Successful calving is measured by the safe delivery of a healthy calf. Any factors that\r cause dystocia (difficult calving) are undesirable; as it will result in weak or dead calves, stress\r dams and a decrease in profit to the rancher. The most often identified reason for dystocia are\r calves that are too large at birth. Ranchers frequently use sires that will result in calves with\r small birth weights that will be born easy. This is especially true when breeding first calving\r heifers. The selection for small calves at birth results not only in less growth in the uterus but\r also less growth after birth, which means less beef to sell. This means ranchers try to balance\r reasonable growth and reasonable calf size when selecting their sires. Much data has been\r published on birth weight and its selection.

BCH (Bnip-2 and Cdc42GAP homology) domains are a novel class of protein-interacting domains. First identified in BNIP2 and in Cdc42GAP, a GTPaseactivating-protein (GAP), canonical BCH domains play important and varied roles in the control of cell morphology and apoptosis induction, amongst others. However, this domain is currently classified under lipid-interacting sec14 domains due to low sequence identity (~20%). Given this intriguing similarity, the relationship between BCH and sec14 domains is worth further study. As BCH/sec14 domains are closely associated with cell signaling and human disease, characterization of these domains and the elucidation of their functional diversity could better aid our understanding of their cellular functions and disease pathogenesis.

This research is mainly in observation with Caenorhabditis elegans ’s genetic mutation caused via nervous system abnormal character. In the study, I the sample have been cultivated purified and add some chemical material EMS to speed up C.elegans mutation. Then based on the character to further analysis what causeof gene deal with mutation and observe the effects in heredity. The research has two stages, on the first stage of study the mainly target is to both search and purify the mutation of C.elegans. The second stage is based on the exploration of mutation’s searching andpurifying. Because the certain mutation bodies aren’t easy to find out, the project is still on progress at the beginning of second stage, and we conclude some heredity special cases in preliminary of study. 這個實驗主要是觀察並針對線蟲因為基因的突變所產生的神經系統異常的變異性狀，在實驗中我先將樣品線蟲培養並純化至一定數量，並加入適當藥劑EMS造成其突變，經篩選並分析此性狀，進而找出造成其突變之基因，以及觀察此性狀對遺傳表現所造成的影響。 該計畫分成兩階段，第一階段的實驗重點是在突變株的搜尋以及純化上，第二階段則是在突變基因的探討上，由於特定突變株的搜尋並非容易，所以目前計畫只進展至第二階段的遺傳實驗初期，對於其遺傳特徵與突變形式上已有了初步的分析，但尚未定位出該基因的位置。

顆粒澱粉可和碘形成藍色錯合物(starch-iodine complex)，我從種子發芽澱粉酶實驗了解澱粉酶(amylase)具糖解酵素活性，會分解澱粉盤上的澱粉，並使澱粉失去與碘形成螺旋狀澱粉-碘錯合物的能力，讓藍色澱粉盤出現透明水解圈。通常澱粉酶之分子結構包含澱粉吸附區及催化區，分別負責吸附大分子或不可溶性澱粉糖(amylose)配體及分解澱粉糖成可溶性小分子糖。本計劃選用分子量、溶解度不同的澱粉糖分別與碘液反應，再加入澱粉酶(glucoamylase)或其澱粉吸附區(starch binding domain, SBD)，發現二者均會破壞澱粉糖與碘結合之螺旋狀結構。分析二者之反應效率，可比較不同蛋白質解開澱粉-碘錯合物之能力。此外，本研究亦比較正常與突變株二聚體澱粉吸附區(SBD dimer)之解開澱粉-碘錯合物反應活性，再查詢文獻、比對不同蛋白質的序列、結構、及功能特性進行綜合分析。根據本研究結果，澱粉吸附蛋白質序列中解開配體螺旋結構最重要的官能基為兩個具有苯環的胺基酸，且二者對於可溶性及不可溶性配體之影響不同。本研究提供科學家了解自然界具備重複澱粉吸附區(tandem repeat)功能的邏輯方法，未來可應用於澱粉代謝、合成相關之蛋白質設計、結構預測、及功能分析。.

CD157 is a 31kDa ectoenzyme that is expressed at elevated levels in patients\r with rheumatoid arthritis (RA). The full function of this protein has yet to be\r determined, and the correlation between CD157 and RA is not well\r understood.

茶品的發酵程度對其香氣與口感有決定性的影響，然而業界卻缺乏相關的客觀指標與其有效的量測方法。茶品發酵的主要化學過程是其兒茶素的氧化與聚合，這些茶單寧將隨著發酵過程逐漸由還原態轉變成氧化態。所以，不同發酵程度的茶，其氧化態單寧與還原態單寧的比將有所不同。這個比值預料可由氧化還原電位間接地量測得知。本研究使用白金絲與銀/氯化銀參考電極為電極對來量測此氧化還原電位。量具則使用具高輸入阻抗(大於十億歐姆)的pH 電極用電位計以取得茶汁中微弱的氧化還原電位訊號，可於十秒內取得高再現性(相對標準差小於3%)的電位訊號所得到的電位訊號與發酵程度呈高度的線性相關(相關係數大於0.9)。Degree of fermentation is pivotal for the aroma and the aftertaste of tea, but an objective scale and efficient evaluating methods are demanded for the important quality factor. Oxidation along with the polymerization of catechins is known to be the major chemical process during tea fermentation; the tea tannins will gradually change from their reduced forms to their oxidized formats. As the consequence, the ratio of oxidized tannins to reduced tannins will differ with the extent of fermentation. The ratio can be measured simply by the redox potential. In the present study, we used a platinum wire and an Ag/AgCl reference electrode as the electrode pair to measure the redox potential. A pH meter with high input impedance (over Giga ohm) served as our voltmeter for the weak voltage signals retrieved from tea infusions. The measurement was quick (less than 10 second) and with good reproducibility (CV0.9).

Entamoeba histolytica is a protozoan parasite known to cause infectious colitis and amoebic dysentery in humans. Its life cycle consist of two parts: the infectious cyst stage and the multiplying trophozoite stage. Epinephrine, a neurotransmitter in vertebrates, is released by the trophozoites during the process of cyst formation. The addition of epinephrine to in vitro cultures of amoebas causes them to encyst, and addition of compounds that prevent epinephrine’s activity inhibits encystations. Therefore, epinephrine plays a critical role in encystation in vitro. An understanding of the molecular intricacies of epinephrine-induced encystations may allow for pharmacological manipulation of epinephrine metabolism to control cyst formation in vitro. Drugs that either prevent cyst formation or induce it before a large amoebic population is present would result in the release of fewer cyst forms of the parasite, reducing parasite transmission from person to person. Although trophozoites release epinephrine, it is no known if E.histolytica synthesizes epinephrine or extracts it from the growth medium. Phenylethanolamine N-methyltransferase(PNMT) is the enzyme that catalyzes production of epinephrine norepinephrine. This study aims to determine the source of epinephrine by determining if E.histolytica contains a PNMT-type enzyme. PNMT amino acid sequences from several higher organisms were compared to identify conserved regions of the enzyme. These conserved amino acid sequences were then used to search for similar sequences in a database containing the recently sequenced amoeba genome. A PNMT-like gene was found in the E.histolytica database and cloned in bacteria. Yeast cells containing the cloned E.histolytica PNMT gene expressed PMT enzyme activity. This suggests that E.histolytica produces its own epinephrine, and is the most evolutionarily ancient eukaryote shown to do so. The use of inhibitors against PNMT activity is under investigation.

The physiochemical properties of fullerenes have aroused wide interest, such as its ability to accept and lose electrons and relatively high reactivity that permit various modes of structural modifications. However, obstacles to further research include its complete lack of solubility in water and low processability.\r This project investigated the morphology and microstructure of a fullerene-grafted polymer as a potential candidate for better and novel systems for drug delivery. In this research, hydrophilic functionalities were introduced to the C60 fullerene by chemical modifications, through the attachment of poly(acrylic acid) (PAA) chain. The objective was to investigate the dynamics and the self-assembly properties of this polymer in aqueous solutions, and the knowledge gained would enhance the development of such system for potential applications in drug delivery and nanotechnology.

L-多巴在酪氨酸酶的作用下，可以轉換成多巴醌，多巴醌繼而自發轉變為黑色素，在此實驗當中，利用帶有青枯病菌的酪胺酸酶基因片段pBAD-518458-bbr的大腸桿菌(E. coli DH5α)將L-多巴轉換成黑色素，然後將微生物去除，只保留黑色素，以避免之後的實驗受到細菌的干擾。 分別將四種金屬離子，Au3+、Cu2+、Co2+、Pd2+ 以1.0 mM的濃度加入黑色素中，使黑色素將金屬離子還原成金屬奈米粒子，形成含有黑色素與金屬奈米粒子的混合溶液。將黑色素去除，避免之後測吸收時受到干擾。再以冷凍乾燥機抽乾乾燥，去除水分以利保存。 將對-硝基苯胺與已乾燥的金屬依照一定濃度加入96孔盤中，再加入硼氫化鈉當作還原劑，也是提供氫陰離子的來源，將對-硝基苯胺還原成對-苯二胺，用Multi-Mode Microplate Reader (BioTeK)測每隔10秒波長385 nm的吸收，再將數據整理成反應速率常數。在四種金屬當中，奈米金的催化速率最佳，所以選擇金屬金以濃度0.1、0.5、1.0 mM作濃度的探討。