一 | RENO, Nev. -- In an ongoing legal battle with the Biden administration over a Nevada lithium mine, environmentalists are poised to return to court with a new approach accusing U.S. wildlife officials of dragging their feet on a year-old petition seeking endangered species status for a tiny snail that lives nearby. The Western Watersheds Project said in its formal notice of intent to sue that the government's failure to list the Kings River pyrg as a threatened or endangered species could push it to the brink of extinction.It says the only place the snail is known to exist is in 13 shallow springs near where Lithium Americas is building its Thacker Pass Mine near the Oregon line.President Joe Biden has made ramped-up domestic production of lithium a key part of his blueprint for a greener future. Worldwide demand for the critical element in the manufacture of electric vehicle batteries is projected to increase six-fold by 2030 compared with 2020.Past lawsuits filed by conservationists and tribes have taken aim — largely unsuccessfully — at the Interior Department’s Bureau of Land Management, which they accused of cutting regulatory corners to expedite approval of the mine itself in 2021.The new approach targets the department's Fish and Wildlife Service, charged with ensuring protection of fish and wildlife habitat surrounding the mine site 200 miles (321 kilometers) northeast of Reno.Western Watersheds Project says groundwater pumping associated with the mine’s 370-foot-deep (113-meter) open pit will reduce or eliminate flows to the springs that support the snails.In the formal 90-day notice of intent to sue sent to Interior Secretary Deborah Haaland last month, they say her agency's failure to make a 12-month finding on the listing petition filed in September 2022 is a violation of the Endangered Species Act.“The Fish and Wildlife Service isn’t supposed to sit on its hands while species are in imminent danger of extinction, but the fact that it hasn’t met the deadlines on the pyrg raises questions about why they might be delaying,” Adam Bronstein, the project's Nevada director, said in a statement.“It would be absolutely unacceptable if the Biden Administration is waiting until it’s too late to save the species so as not to interrupt the construction of a lithium mine," he said.Interior Department spokeswoman Melissa Schwartz said in an email Thursday the department had no comment on the group's intent to sue.Western Watersheds Project said time is of the essence because the snails were imperiled even before any new mining was contemplated due to livestock grazing, round-building and, increasingly, the anticipated impacts of climate change.“The species has no regulatory protection whatsoever ... because it is not an endangered species, or even a Bureau of Land Management-listed Sensitive species, and has no state law protections," the notice said.Conservationists and tribal lawyers claimed a partial victory last year when U.S. District Judge Miranda Du concluded the bureau failed to fully comply with new interpretations of the 1872 Mining Law. But she stopped short of blocking the project, allowing construction to begin as the bureau shored up plans for disposal of waste rock.The opponents appealed, but the 9th U.S. Circuit Court of Appeals upheld Du's ruling in July.The tiny snail’s shell is less than 2 millimeters (0.08 inches) tall. By comparison, a U.S. nickel coin is 1.95 millimeters thick. They've managed to survive in isolated springs, which are remnants of extensive waterways that have covered what is now dry land only to recede many times over the last 2 million years, the listing petition said.The project says three of the springs are within a 1-mile (1.6-kilometer) buffer zone, the bureau established in its review of potential impacts of a 10-foot (3-meter) drawdown of the groundwater table, and the rest are less than 4 miles (4.8 kilometers) away."As drought frequency increases with climate change, the Kings River pyrg will be at high risk of extinction,” the letter to Haaland said. It notes that the Nevada Department of Wildlife considers the pyrg “extremely vulnerable to climate change.”Lithium Americas had no comment on the notice of intent to sue the Fish and Wildlife Service, spokesman Tim Crowley said. The company said when the listing petition was filed last year that it's done extensive work to design a project that avoids impacts to the springs.The Bureau of Land Management said earlier its environmental review of the project that it didn't detect any of the snails "within the direct footprint of the project or any area likely to be adversely affected by the project.”。
二 | 由麦凯尔维工程学院机械工程和材料科学的Lilyan & E. Lisle Hughes教授Srikanth Singamaneni和Singamaneni实验室的前博士后Anushree Seth领导的研究人员开发了FluoroDOT测定法。该研究最近发表在《细胞报告方法》上。
三 | 这种高度敏感的检测方法能够在大约30分钟内看到并测量单个细胞分泌的蛋白质。树突状细胞在不同时间点分泌蛋白1（TNFa，红色显示）和蛋白2（IL-6，黄色显示）的FluoroDOT检测图像（从左到右。未受刺激、受刺激30分钟、1小时、2小时和3小时）。细胞的细胞核以蓝色显示。白色箭头突出显示了只分泌一种蛋白质或两种蛋白质的不同数量和不同时间点的细胞。图像: Srikanth Singamaneni/华盛顿大学与来自其他大学和华盛顿大学医学院的科学家一起发现FluoroDOT检测法用途广泛，价格低廉，可适应任何实验室环境，并且有可能比目前常用的检测法更全面地了解这些蛋白质。生物医学研究人员通过分泌蛋白来了解细胞间通讯、细胞信号传导、激活和炎症等方面的信息，但目前的方法在敏感性方面受到限制，而且可能需要24小时的繁琐处理流程。质子-荧光是Singamaneni实验室创造的一种质子增强的纳米标签，比传统的荧光标签亮16000倍，信噪比超过30倍，这是FluoroDOT检测法与其他检测法的区别所在。Singamaneni说："质子-荧光剂是由金属纳米粒子组成的，它作为天线拉入光线并增强分子荧光团的荧光发射，从而使它成为一种超亮的纳米粒子。
四 | "这种质子-荧光的超亮发射使用户能够看到极少量的分泌蛋白，这是他们在现有测定法中无法做到的，加上利用定制的算法，用每个簇或斑点的颗粒数量或点状图案，实现以数字方式测量高分辨率信号。此外，它不需要特殊设备。Singamaneni和他的合作者于2020年在《自然-生物医学工程》杂志上首次发表了他们的等离子体氟化物工作。这项正在申请专利的质子荧光技术由圣路易斯华盛顿大学技术管理办公室授权给Auragent生物科学有限公司。"使用一个简单的荧光显微镜，我们能够同时对一个细胞以及它周围分泌的蛋白质的空间分布进行成像，"Seth说，他作为Singamaneni实验室的博士后学者从事这个项目，并作为Auragent生物科学公司的首席科学家（细胞应用）继续从事这项工作。"我们看到不同细胞类型的有趣分泌模式。这种检测方法还能同时观察来自单个细胞的两种类型的蛋白质。当多个细胞受到相同的刺激时，我们可以将同时分泌两种蛋白质的细胞与只分泌一种蛋白质或根本不分泌的细胞区分开来。
五 | "为了验证这项技术，该团队使用了从人类和小鼠细胞中分泌的蛋白质，包括感染了结核分枝杆菌的免疫细胞。合作者和共同作者之一，医学博士Jennifer A. Philips，医学和分子微生物学系的Theodore和Bertha Bryan教授，以及医学院传染病科的共同主任，已经在她的实验室中使用了FluoroDOT测定。Philips举例说："当结核分枝杆菌感染免疫细胞时，这些细胞通过分泌重要的免疫蛋白（称为细胞因子）做出反应。但不是所有的细胞都以同样的方式对感染做出反应。FluoroDOT测定使我们能够看到群体中的单个细胞如何对感染做出反应--看到哪些细胞正在分泌以及朝哪个方向分泌。这在旧技术中是不可能的。
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