CO2浓度升高和施氮条件下小麦根际呼吸对土壤呼吸的贡献

依托FACE技术平台,采用稳定13C同位素技术,通过将小麦(C3作物)种植于长期单作玉米(C4作物)的土壤上,研究了大气CO2浓度升高和不同氮肥水平对土壤排放CO2的δ13C值及根际呼吸的影响.结果表明:种植小麦后土壤排放CO2的δ13C值随作物生长逐渐降低,CO2浓度升高200 μmol·mol-1显著降低了孕穗、抽穗期(施氮量为250 kg·hm-2,HN)与拔节、孕穗期(施氮量为150 kg·hm-2,LN)土壤排放CO2的δ13C值,显著提高了孕穗、抽穗期的根际呼吸比例.拔节至成熟期,根际呼吸占土壤呼吸的比例在高CO2浓度下为24％～48％ (HN)和21％ ～48％ (LN),在正常CO2浓度下为20％ ～36％ (HN)和19％～32％(LN).不同CO2浓度下土壤排放CO2的δ13C值和根际呼吸对氮肥增加的响应不同,CO2浓度与氮肥用量在拔节期对根际呼吸的交互效应显著.     

CO2浓度和温度升高对红桦根际微生物的影响

应用自控、封闭、独立的生长室系统,研究升高的大气CO2浓度(环境CO2浓度 350(±25)μmol.mol-1,EC)和温度(环境温度 2.0(±0.5)℃,ET)及其交互作用(ECT)对不同栽植密度条件下红桦根际土壤可培养微生物数量的影响。结果表明:(1)EC显著增加了高密度条件下根际细菌数量;在整个生长季中,最大的根际细菌数量增加出现在7月份;而EC对低密度处理的根际细菌数量影响不显著。除了5月和6月份,ET在其余月份均显著增加了根际细菌数量,但是与密度处理没有有意义的相关;ECT对高低密度处理的根际细菌数量均未产生有统计意义的影响。(2)EC对低密度条件下的根际放线菌数量有显著增加,而对高密度条件下的根际放线菌数量无显著影响;ET和ECT对高低密度条件下的根际放线菌数量均未产生有统计意义的影响。(3)EC和ET对高低密度条件下的根际真菌数量无显著增加,而ECT显著增加了根际真菌数量。     

大气CO2浓度升高对不同施氮土壤酶活性的影响

利用中国唯一的无锡FACE（Free-air CO2 enrichment,开放式空气CO2浓度升高）平台,研究了大气CO2浓度升高对土壤β-葡糖苷酶、转化酶、脲酶、酸性磷酸酶、-氨基葡糖苷酶的影响。研究发现,不同氮肥处理下大气CO2浓度升高对某些土壤酶活性的影响不同。在低氮施肥处理中,大气CO2浓度升高显著降低-葡糖苷酶活性,但是在高氮施肥处理下,大气CO2浓度升高显著增加β-葡糖苷酶活性。在低氮和常氮施肥处理中大气CO2浓度升高显著增加了土壤脲酶活性,但在高氮水平下影响不显著。在低氮、常氮施肥处理中,大气CO2浓度升高对土壤酸性磷酸酶活性没有影响,而在高氮施肥处理中显著增强了土壤中磷酸酶活性。大气CO2浓度升高对土壤转化酶活性和-氨基葡糖苷酶的活性有增加趋势,但影响不显著。研究还发现,在不同的CO2浓度下,土壤酶活性对不同氮肥处理的响应也不同。在正常CO2浓度下,土壤中β-葡糖苷酶活性随着氮肥施用量的增加而降低,而在大气CO2浓度升高条件下,却随着氮肥施用量的增加而增加。在大气CO2浓度升高条件下,高氮施肥显著增加了转化酶和酸性磷酸酶活性,而在正常CO2浓度下,影响不显著。在大气CO2浓度升高条件下,氮肥处理对脲酶活性的影响不大,但在正常CO2浓度下,脲酶活性随着氮肥施用量的增加而增加。氮肥对β-氨基葡糖苷酶活性的影响不明显。     

短期CO2浓度升高和干旱胁迫对白羊草土壤碳氮和微生物根际效应的影响

采用人工气候室和盆栽控水试验研究黄土丘陵区典型草本植物白羊草在倍增CO₂浓度(800 μmol·mol^-1)下和充分供水(75%～80%的田间持水量)、轻度干旱胁迫(55%～60%的田间持水量)和重度干旱胁迫(35%～40%的田间持水量)下根际和非根际土壤碳氮含量和微生物群落结构及其根际效应.结果表明: CO₂浓度升高和干旱胁迫对白羊草根际和非根际土壤有机碳、全氮和水溶性有机碳(DOC)含量及其根际效应均无显著影响.轻度干旱胁迫下CO₂浓度升高显著促进了根际土壤水溶性有机氮(DON)的消耗,导致DOC/DON升高,提高了DON的负根际效应和DOC/DON的正根际效应.干旱胁迫和CO₂浓度升高对土壤总磷脂脂肪酸(总PLFA)和细菌PLFA的根际效应无显著影响.CO₂浓度升高条件下干旱胁迫显著提高了根际土壤G⁺/G^- PLFA,降低了非根际土壤G⁺/G^- PLFA,导致其根际效应显著提高,表明根际微生物群落由自养微生物群落向异养微生物群落的转变.     

冬小麦旺盛生长期间CO2浓度升高对根际呼吸的影响

依托FACE（free air carbon dioxide enrichment）技术平台,利用阻断根法,采用H6400红外气体分析仪（IRGA）-田间原位测定的方法,研究了大气CO2浓度升高和不同氮肥水平对水稻／小麦轮作制中冬小麦旺盛生长期间根际呼吸的影响。结果表明,在整个测定期间,大气CO2浓度升高增强了根际呼吸速率,提高了根际呼吸排放量。在高N和低N处理中,高CO2浓度下的根际呼吸总排放量分别比Ambient极显著增加117．0％和90．8％。根际呼吸速率在孕穗初期达到最大值;使根际呼吸在土壤呼吸中的比重由24．5％（LN）～26．7（HN）提高到39．8％（LN）～47．1％（HN）。CO2浓度升高与氮肥用量对根际呼吸产生交互效应。表明大气CO2浓度升高将加快土壤向大气的CO2排放,结果将有助于评价未来高CO2浓度背景下农田生态系统土壤碳的固定潜力。     

短期CO2浓度升高和干旱胁迫对白羊草土壤碳氮和微生物根际效应的影响

采用人工气候室和盆栽控水试验研究黄土丘陵区典型草本植物白羊草在倍增CO₂浓度(800 μmol·mol^-1)下和充分供水(75%～80%的田间持水量)、轻度干旱胁迫(55%～60%的田间持水量)和重度干旱胁迫(35%～40%的田间持水量)下根际和非根际土壤碳氮含量和微生物群落结构及其根际效应.结果表明: CO₂浓度升高和干旱胁迫对白羊草根际和非根际土壤有机碳、全氮和水溶性有机碳(DOC)含量及其根际效应均无显著影响.轻度干旱胁迫下CO₂浓度升高显著促进了根际土壤水溶性有机氮(DON)的消耗,导致DOC/DON升高,提高了DON的负根际效应和DOC/DON的正根际效应.干旱胁迫和CO₂浓度升高对土壤总磷脂脂肪酸(总PLFA)和细菌PLFA的根际效应无显著影响.CO₂浓度升高条件下干旱胁迫显著提高了根际土壤G⁺/G^- PLFA,降低了非根际土壤G⁺/G^- PLFA,导致其根际效应显著提高,表明根际微生物群落由自养微生物群落向异养微生物群落的转变.     

CO2浓度增加对长白山森林土壤甲烷氧化影响

大气CO2浓度升高可能对森林土壤的甲烷(CH4)氧化速率产生影响.本文采用开顶箱技术,对连续6年高浓度CO2(500 μmol·mol-1)处理的长白山森林典型树种蒙古栎树下土壤CH4氧化速率进行研究,并利用CH4氧化菌的16S rRNA特异性引物以及CH4单加氧酶功能基因引物分析了土壤中CH4氧化菌的群落结构与数量.结果表明:CO2浓度增高后,生长季土壤甲烷氧化量与对照和裸地相比分别降低了4％和22％;基于16S rRNA特异性引物的DGGE分析表明,CO2浓度增高导致两类甲烷氧化菌的多样性指数降低;CO2浓度增高对土壤中Ⅰ类甲烷氧化菌数量无显著影响,而使土壤中Ⅱ类甲烷氧化菌数量显著减少,功能基因pmoA拷贝数与对照和裸地相比分别降低了15％和46％.CO2浓度增高导致森林土壤甲烷氧化菌数量与活性降低,土壤含水量的增加可能是导致这一现象的主要原因.     

大气CO2浓度升高对稻麦根系周围土壤C、P、K的影响

利用FACE(free air carbon dioxide enrichment)技术平台,在两种氮肥施用(低氮,LN和常规氮,NN)水平下,研究CO2浓度升高对水稻和小麦收获后根际和非根际土壤可溶性碳、有机磷、速效磷和速效钾的影响.结果表明,相对于对照CO2浓度处理,高CO2浓度处理在显著增加作物生物量的前提下,土壤速效磷和速效钾不但没有降低反而增加,增加幅度小麦季大于水稻季,根际大于非根际;水稻季土壤可溶性碳含量增加,且NN水平下水稻和小麦季进入土壤的可溶性碳增加,导致土壤有机磷降低幅度低于LN水平,且水稻季根际土壤大于非根际土壤,有机磷的降低是保证有效磷升高的一个重要因素,增加氮肥施用将有利于土壤有机磷的增加,对维持土壤磷的供给有积极作用,有利于作物对高CO2浓度的持续响应.     

大气二氧化碳浓度升高对植物根际微生物及菌根共生体的影响

综述了大气CO2浓度升高条件下,植物根际土壤环境、根际土壤微生物和植物菌根形成的变化趋势等方面的研究进展,CO2浓度升高,运转到根系的碳水化合物增加,根际环境、根际微生物活性、微生物群落结构以及菌根共生体的形成发生变化．提出在CO2浓度升高条件下,根际微生物和菌根真菌群落的变化对植物群落和陆地生态系统碳动态的调节是今后的研究趋向。     

氮素添加和CO2浓度升高对白羊草根际和非根际土壤水溶性有机碳、氮的影响

采用盆栽控制试验对黄土丘陵区白羊草在不同CO₂浓度(400和800 μmol·mol^-1)和施氮水平(0、2.5、5.0 g N·m^-2·a^-1)条件下根际和非根际土壤水溶性有机碳(DOC)和水溶性有机氮(DON)的变化特征进行研究.结果表明: CO₂浓度升高对白羊草根际和非根际土壤DOC、水溶性总氮(DTN)、DON、水溶性铵态氮(NH₄⁺-N)、水溶性硝态氮(NO₃^--N)含量均无显著影响.施氮显著提高了根际和非根际土壤DTN、NO₃^--N含量和根际土壤DON含量,显著降低了根际土壤DOC/DON.在各处理条件下,根际土壤DTN、NO₃^--N和DON含量均显著低于非根际土壤,根际土壤DOC/DON显著高于非根际土壤.短期CO₂浓度升高对黄土丘陵区土壤水溶性有机碳、氮含量无显著影响,而氮沉降的增加在一定程度上改善了土壤中水溶性氮素缺乏的状况,但并不足以满足植被对水溶性氮素的需求.     

Preparation of hair for measurement of elements by inductively coupled plasma-mass spectrometry (ICP-MS)

The preparation of hair for the determination of elements is a critical component of the analysis procedure. Open-beaker, closedvessel microwave, and flowthrough microwave digestion are methods that have been used for sample preparation and are discussed. A new digestion method for use with inductively coupled plasma-mass spectrometry (ICP-MS) has been developed. The method uses 0.2 g of hair and 3 mL of concentrated nitric acid in an atmospheric pressurelow-temperature microwave digestion (APLTMD) system. This preparation method is useful in handling a large numbers of samples per day and may be adapted to hair sample weights ranging from 0.08 to 0.3 g. After digestion, samples are analyzed by ICP-MS to determine the concentration of Li, Be, B, Na, Mg, Al, P, S, K, Ca, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ge, As, Se, Rb, Sr, Zr, Mo, Pd, Ag, Cd, Sn, Sb, I, Cs, Ba, Pt, Au, Hg, Tl, Pb, Bi, Th, and U. Benefits of the APLTMD include reduced contamination and sample handling, and increased precision, reliability, and sample throughput.     

外泌体在白血病中的重要调控作用

目前,白血病复发是患者死亡的主要原因之一。肿瘤细胞和微环境的相互作用,以及隐匿在骨髓中的肿瘤干细胞,促进了白血病的复发和向淋巴组织的转移,因此白血病的治疗、转移和复发问题受到广泛关注。外泌体是由绝大多数细胞分泌的双层脂质膜囊泡,可以调控细胞间的交流和信息传递。在白血病细胞、基质细胞和内皮细胞之间的相互联系中都涉及到外泌体,白血病细胞来源的外泌体存在于白血病患者的血浆中,能把其携带的白血病相关抗原及微小RNA呈递给靶细胞,促进白血病肿瘤细胞的增殖,有助于肿瘤细胞实现免疫逃避,保护白血病细胞抵抗化疗药物导致的细胞毒性作用,促进血管生成及肿瘤细胞的迁移。因此,外泌体与白血病的转移、治疗及预后密切相关,可以用来检测和监测白血病的进展。本文综述了外泌体的来源、形成与分泌机制,以及外泌体在白血病发生前、发展中、预后和免疫治疗中所扮演的重要角色。     

日本北海道虻类雌性外生殖器形态(双翅目：虻科)(英文)

【目的】利用日本北海道虻类评估和验证外生殖器在分类学上的意义。【方法】将虻类成虫标本浸渍在生理盐水中并置于双目显微镜下通过针和镊子在培养皿中进行解剖并绘图,观察第9背板、第10背板、尾叶、第8腹板、受精囊、受精囊管及生殖叉器的形态特征。【结果】在日本北海道共记录了虻科(Tabanidae) 3亚科7属38种。我们观察并描述了3亚科其中的6属24种的雌性外生殖器的主要特征。亚科之间存在明显差异;然而在一般情况下属之间很难建立一种方法来确定共同点;种之间只有在斑虻属Chrysops中有相似之处,其他属中则比较多样化。因此,亚科鉴定根据第9背板、第8腹板及受精囊足以进行区分,属及种鉴定需要结合第9背板、第10背板、尾叶、第8腹板、受精囊、受精囊管及生殖叉器各自的特征组合在一起才能区分开来。我们也制作了虻类外生殖器的检索表。【结论】和许多其他昆虫一样,外生殖器是虻科的重要分类特征,对于促进分类学和系统学的发展具有重要意义。本研究首次对分布在日本北海道的虻科雌性外生殖器进行了系统研究。     

Ecological importance of sedges: a survey of the Australasian Cyperaceae genus Lepidosperma

Background

Sedges (Cyperaceae) form an important ecological component of many ecosystems around the world. Sword and rapier sedges (genus Lepidosperma) are common and widespread components of the southern Australian and New Zealand floras, also occurring in New Caledonia, West Papua, Borneo, Malaysia and southern China. Sedge ecology is seldom studied and no comprehensive review of sedge ecology exists. Lepidosperma is unusual in the Cyperaceae with the majority of species occurring in dryland habitats.

Scope

Extensive review of ecological literature and field observations shows Lepidosperma species to be important components of many ecosystems, often dominating understorey and sedge-rich communities. For the first time, a detailed ecological review of a Cyperaceae genus is presented.

Conclusions Lepidosperma

species are long-lived perennials with significant abundance and persistence in the landscape. Speciation patterns in the genus are of considerable interest due to complex biogeographical patterns and a high degree of habitat specificity. Potential benefits exist for medicinal products identified from several Lepidosperma species. Over 178 organisms, including 26 mammals, 42 birds, six reptiles, five amphibians, eight arachnids, 75 insects, three crustaceans and 13 fungi, are found to be dependent on, or making use of, Lepidosperma species. A significant relationship exists between Lepidosperma species and the moth genus Elachista. Implications for the conservation and ecology of both sedges and associated species are discussed.     

Literature based species occurrence data of birds of northeast India

The northeast region of India is one of the world's most significant biodiversity hotspots. One of the richest bird areas in India, it is an important route for migratory birds and home to many endemic bird species. This paper describes a literature-based dataset of species occurrences of birds of northeast India. The occurrence records documented in the dataset are distributed across eleven states of India, viz.: Arunachal Pradesh, Assam, Bihar, Manipur, Meghalaya, Mizoram, Nagaland, Sikkim, Tripura, Uttar Pradesh and West Bengal. The geospatial scope of the dataset represents 24 to 29 degree North latitude and 78 to 94 degree East longitude, and it comprises over 2400 occurrence records. These records have been collated from scholarly literature published between1915 and 2008, especially from the Journal of the Bombay Natural History Society (JBNHS). The temporal scale of the dataset represents bird observations recorded between 1909 and 2007. The dataset has been developed by employing MS Excel. The key elements in the database are scientific name, taxonomic classification, temporal and geospatial details including geo-coordinate precision, data collector, basis of record and primary source of the data record. The temporal and geospatial quality of more than 50% of the data records has been enhanced retrospectively. Where possible, data records are annotated with geospatial coordinate precision to the nearest minute. This dataset is being constantly updated with the addition of new data records, and quality enhancement of documented occurrences. The dataset can be used in species distribution and niche modeling studies. It is planned to expand the scope of the dataset to collate bird species occurrences across the Indian peninsula.     

Characterization of citrus pectin samples extracted under different conditions: influence of acid type and pH of extraction

Background and Aims

Pectin is a complex macromolecule, the fine structure of which is influenced by many factors. It is used as a gelling, thickening and emulsifying agent in a wide range of applications, from food to pharmaceutical products. Current industrial pectin extraction processes are based on fruit peel, a waste product from the juicing industry, in which thousands of tons of citrus are processed worldwide every year. This study examines how pectin components vary in relation to the plant source (orange, lemon, lime, grapefruit) and considers the influence of extraction conditions on the chemical and macromolecular characteristics of pectin samples.

Methods

Citrus peel (orange, lemon, lime and grapefruit) from a commercial supplier was used as raw material. Pectin samples were obtained on a bulk plant scale (kilograms; harsh nitric acid, mild nitric acid and harsh oxalic acid extraction) and on a laboratory scale (grams; mild oxalic acid extraction). Pectin composition (acidic and neutral sugars) and physicochemical properties (molar mass and intrinsic viscosity) were determined.

Key Results

Oxalic acid extraction allowed the recovery of pectin samples of high molecular weight. Mild oxalic acid-extracted pectins were rich in long homogalacturonan stretches and contained rhamnogalacturonan I stretches with conserved side chains. Nitric acid-extracted pectins exhibited lower molecular weights and contained rhamnogalacturonan I stretches encompassing few and/or short side chains. Grapefruit pectin was found to have short side chains compared with orange, lime and lemon. Orange and grapefruit pectin samples were both particularly rich in rhamnogalacturonan I backbones.

Conclusions

Structural, and hence macromolecular, variations within the different citrus pectin samples were mainly related to their rhamnogalacturonan I contents and integrity, and, to a lesser extent, to the length of their homogalacturonan domains.     

The Rumen Ciliate Fauna of Domestic Sheep (Ovis ammon aires) from the Turkish Republic of Northern Cyprus

ABSTRACT. Concentration and composition of ciliate protozoa in the families Ophryoscolecidae and Isotrichidae were determined in rumen contents of domestic sheep ( Ovis ammon aries ) from Cyprus. A total of five genera of Ophryoscolecidae were identified, Metadinium, Enoploplastron, Polyplastron, Epidinium , and Ophryoscolex , which included six species: Metadinium affine, Enoploplastron triloricatum, Polyplastron multivesiculatum, Epidinium ecaudatum, Epidinium graini, and Ophryoscolex purkynjei. Eight separate forms of Epidinium were identified ( E. ecaudatum f. ecaudatum, E, e. f. caudatum, E. e. f. bicaudatum, E. e. f. tricaudatum, E. e. f. quadricaudatum, E. graini f. graini, E. g. f. caudatricoronatum , and E. g. f. caudaquadricoronatum ), along with five forms of Ophryoscolex purkynjei (O. p. f. purkynjei, O. p. f. bifidobicinctus, O. p. f. bifidoquadricinctus, O. p. f. bicoronatus, O. p. f. tricoronatus , and O. p. f. quadricoronatus). Three species of Isotrichidae were observed, Isotricha intestinalis, I. prostoma , and Dasytricha ruminantium. This study reports new host records for three forms of Epidinium graini and Ophryoscolex purkynjei f. bifidobicinctus. The rumen fauna in the family Ophryoscolecidae from Cypriote domestic sheep appear to have limited diversity compared to those from Turkish and Far Eastern (Chinese/Japanese) sheep, while they are more diverse than those found in Western European (Scottish) and North American (Canadian/Alaskan) sheep.     

Micromanipulation of Gene Expression in the Adult Zebrafish Brain Using Cerebroventricular Microinjection of Morpholino Oligonucleotides

Manipulation of gene expression in tissues is required to perform functional studies. In this paper, we demonstrate the cerebroventricular microinjection (CVMI) technique as a means to modulate gene expression in the adult zebrafish brain. By using CVMI, substances can be administered into the cerebroventricular fluid and be thoroughly distributed along the rostrocaudal axis of the brain. We particularly focus on the use of antisense morpholino oligonucleotides, which are potent tools for knocking down gene expression in vivo. In our method, when applied, morpholino molecules are taken up by the cells lining the ventricular surface. These cells include the radial glial cells, which act as neurogenic progenitors. Therefore, knocking down gene expression in the radial glial cells is of utmost importance to analyze the widespread neurogenesis response in zebrafish, and also would provide insight into how vertebrates could sustain adult neurogenesis response. Such an understanding would also help the efforts for clinical applications in human neurodegenerative disorders and central nervous system regeneration. Thus, we present the cerebroventricular microinjection method as a quick and efficient way to alter gene expression and neurogenesis response in the adult zebrafish forebrain. We also provide troubleshooting tips and other useful information on how to carry out the CVMI procedure.     

Design and Use of Multiplexed Chemostat Arrays

Chemostats are continuous culture systems in which cells are grown in a tightly controlled, chemically constant environment where culture density is constrained by limiting specific nutrients.^1,2 Data from chemostats are highly reproducible for the measurement of quantitative phenotypes as they provide a constant growth rate and environment at steady state. For these reasons, chemostats have become useful tools for fine-scale characterization of physiology through analysis of gene expression^3-6 and other characteristics of cultures at steady-state equilibrium.⁷ Long-term experiments in chemostats can highlight specific trajectories that microbial populations adopt during adaptive evolution in a controlled environment. In fact, chemostats have been used for experimental evolution since their invention.⁸ A common result in evolution experiments is for each biological replicate to acquire a unique repertoire of mutations.^9-13 This diversity suggests that there is much left to be discovered by performing evolution experiments with far greater throughput. We present here the design and operation of a relatively simple, low cost array of miniature chemostats—or ministats—and validate their use in determination of physiology and in evolution experiments with yeast. This approach entails growth of tens of chemostats run off a single multiplexed peristaltic pump. The cultures are maintained at a 20 ml working volume, which is practical for a variety of applications. It is our hope that increasing throughput, decreasing expense, and providing detailed building and operation instructions may also motivate research and industrial application of this design as a general platform for functionally characterizing large numbers of strains, species, and growth parameters, as well as genetic or drug libraries.     

A Microplate Assay to Assess Chemical Effects on RBL-2H3 Mast Cell Degranulation: Effects of Triclosan without Use of an Organic Solvent

Mast cells play important roles in allergic disease and immune defense against parasites. Once activated (e.g. by an allergen), they degranulate, a process that results in the exocytosis of allergic mediators. Modulation of mast cell degranulation by drugs and toxicants may have positive or adverse effects on human health. Mast cell function has been dissected in detail with the use of rat basophilic leukemia mast cells (RBL-2H3), a widely accepted model of human mucosal mast cells^3-5. Mast cell granule component and the allergic mediator β-hexosaminidase, which is released linearly in tandem with histamine from mast cells⁶, can easily and reliably be measured through reaction with a fluorogenic substrate, yielding measurable fluorescence intensity in a microplate assay that is amenable to high-throughput studies¹. Originally published by Naal et al.¹, we have adapted this degranulation assay for the screening of drugs and toxicants and demonstrate its use here.Triclosan is a broad-spectrum antibacterial agent that is present in many consumer products and has been found to be a therapeutic aid in human allergic skin disease^7-11, although the mechanism for this effect is unknown. Here we demonstrate an assay for the effect of triclosan on mast cell degranulation. We recently showed that triclosan strongly affects mast cell function². In an effort to avoid use of an organic solvent, triclosan is dissolved directly into aqueous buffer with heat and stirring, and resultant concentration is confirmed using UV-Vis spectrophotometry (using ε₂₈₀ = 4,200 L/M/cm)¹². This protocol has the potential to be used with a variety of chemicals to determine their effects on mast cell degranulation, and more broadly, their allergic potential.