The regulation of reactive oxygen species such as superoxide and nitric oxide is crucial in biology, affecting metabolism and signaling pathways. Researchers have developed two bioluminescent probes, $\mathbf{A}$ and $\mathbf{B}$, which specifically react with superoxide and nitrogen oxides, respectively.  

<MultiModal>![The image shows two molecules separated by a horizontal line. The upper molecule $\mathbf{A}$ has the SMILES formula: O=C(N(CCN(C)C(OC1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1)=O)C)OC4=CC(OC(C)=O)=C(OC(C)=O)C=C4; while the lower molecule $\mathbf{B}$ has the SMILES formula: S=C(NNC(N(C1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1)C)=O)NC4=CC=C([N+]([O-])=O)C=C4](/media/uploads/04631fe2-d200-41af-bc97-0cd8d083c73e.png)</MultiModal>  

By analyzing the structures of the two compounds, determine which of the following options best aligns with the question's intent.
超氧化物和一氧化氮等活性氧的调节在生物学中至关重要，影响新陈代谢和信号通路。研究者们开发了两种生物荧光探针$\mathbf{A}$、$\mathbf{B}$，分别和超氧化物及一氧化氮发生特异的反应。

<MultiModal>![图中上下两个分子被一条横线分开，上方的$\mathbf{A}$分子对应的SMILES式为：O=C(N(CCN(C)C(OC1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1)=O)C)OC4=CC(OC(C)=O)=C(OC(C)=O)C=C4；而下方的$\mathbf{B}$分子对应的SMILES式为：S=C(NNC(N(C1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1)C)=O)NC4=CC=C([N+]([O-])=O)C=C4](/media/uploads/04631fe2-d200-41af-bc97-0cd8d083c73e.png)</MultiModal>

通过对两种物质结构的分析，判断下面选项哪一个是最贴合题意的。
Both molecules $\mathbf{A}$ and $\mathbf{B}$ can immediately react with superoxide or nitrogen oxides upon entering the cell due to their meticulously optimized structural properties.
Due to the structural properties of molecules $\mathbf{A}$ and $\mathbf{B}$, neither can react immediately with superoxide or nitrogen oxides after entering the cell.
After interacting with superoxide, molecule $\mathbf{A}$ forms a quinone and establishes a large and stable conjugated system with other groups, thereby directly generating a visible fluorescence signal.
The $\mathbf{B}$ molecule interacts with nitrogen oxides due to its thiourea-based reactive group, which captures nitric oxide by leveraging the coordination between metal ions (e.g., $\mathrm{Hg}^{2+}$) and nitric oxide, owing to its ability to bind heavy metal ions.
Whether it is $\mathbf{A}$ or $\mathbf{B}$, no biological enzymes are required for fluorescence presentation.
$\mathbf{A}$ requires the involvement of biological enzymes before exhibiting fluorescence, while $\mathbf{B}$ does not require any biological enzyme participation before fluorescing.
Before $\mathbf{B}$ exhibits fluorescence, the involvement of biological enzymes is required, whereas $\mathbf{A}$ does not require any biological enzyme participation before exhibiting fluorescence.
All of the above options are correct.
None of the above options is correct
$\mathbf{A,B}$两种分子均可以因其精心优良的结构性质，在进入细胞之后立刻和超氧化物或一氧化氮进行反应。
因$\mathbf{A,B}$两种分子的结构性质，在进入细胞之后均不可以立刻和超氧化物或一氧化氮进行反应。
$\mathbf{A}$分子和超氧化物作用后形成醌，和其他集团形成庞大而稳定的共轭体系，因此直接产生可见的荧光信号。
$\mathbf{B}$分子和一氧化氮作用后是由于其具有基于硫代氨脲的反应基团，因其能够结合重金属离子如$\mathrm{Hg}^{2+}$等的性质，利用金属离子和一氧化氮的配位作用捕捉一氧化氮。
不论是$\mathbf{A}$还是$\mathbf{B}$，呈现荧光之前中均不需要任何生物酶参与。
$\mathbf{A}$呈现荧光之前中需要生物酶参与，$\mathbf{B}$呈现荧光之前不需要任何生物酶参与。
$\mathbf{B}$呈现荧光之前中需要生物酶参与，$\mathbf{A}$呈现荧光之前不需要任何生物酶参与。
其他的选项均正确。
其他的选项均不正确
The researchers developed two bioluminescent probes (A and B) for the selective detection of superoxide and nitric oxide in living organisms. The "bioluminescent probes" used mimic the luminescent mechanism of fireflies in living organisms, where the luminescence process requires the participation of luciferase.

<MultiModal>![The mechanism of fluorescence generation for probes A and B proposed in the article. From top to bottom and left to right, the corresponding SMILES are:

O=C(N(CCN(C)C(OC1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1)=O)C)OC4=CC(OC(C)=O)=C(OC(C)=O)C=C4

OC1=C(O)C=CC(OC(N(CCN(C)C(OC2=CC(SC(C3=N[C@H](C(OC)=O)CS3)=N4)=C4C=C2)=O)C)=O)=C1

O=C1C(C=CC(OC(N(CCN(C)C(OC2=CC(SC(C3=N[C@H](C(OC)=O)CS3)=N4)=C4C=C2)=O)C)=O)=C1)=O

OC1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1

S=C(NNC(N(C1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1)C)=O)NC4=CC=C([N+]([O-])=O)C=C4

O=C(OC)[C@H](CS1)N=C1C2=NC3=C(S2)C=C(NC4=NN=C(NC5=CC=C([N+]([O-])=O)C=C5)O4)C=C3

NC1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1](/media/uploads/4e683c29-fa56-4878-9934-cf531a51b78d.png)</MultiModal>

<Checkpoint>[Fluorescence generation utilizes the luciferin system](0.5)</Checkpoint>

Analysis of the structures reveals that both probe molecules share a common core skeleton with the SMILES formula `O=C(OC)[C@H](CS1)N=C1C2=NC(C=C3)=C(S2)C=C3OC`. This is a derivative of D-luciferin methyl ester, where D-luciferin is the substrate in the firefly luminescence system. It undergoes oxidation catalyzed by ATP and luciferase to produce fluorescence.

<MultiModal>![SMILES of the luciferin methyl ester group: O=C(OC)[C@H](CS1)N=C1C2=NC(C=C3)=C(S2)C=C3OC](/media/uploads/ef9f7eda-5160-4ee0-93f0-83c77f770b2c.png)</MultiModal>

<Checkpoint>[Both probes possess the luciferin skeleton](0.5)</Checkpoint>

For $\mathbf{A}$, the reaction process proposed in the article is as follows:

Step 1: Deprotection. The acetyl group is typically hydrolyzed by esterases in living organisms, generating the corresponding phenolic hydroxyl group. This is an enzymatic reaction.

<Checkpoint>[Probe $\mathbf{A}$ first requires hydrolysis by esterase](0.5)</Checkpoint>

Step 2: Reaction with superoxide. After esterase hydrolysis, a catechol structure is formed. This structure is first oxidized to a semiquinone and then further oxidized to an ortho-quinone. Although it forms a certain conjugated structure with the adjacent system, this does not emit fluorescence.

<Checkpoint>[Probe $\mathbf{A}$ undergoes an ortho-quinone intermediate](0.5)</Checkpoint>

Step 3: Hydrolysis. The ortho-quinone is an electron-deficient system and is rapidly hydrolyzed, followed by the rapid hydrolysis of the phenolic ester on the right side of the D-luciferin methyl ester derivative, leaving behind the D-luciferin methyl ester portion.

Step 4: Hydrolysis by esterase. D-luciferin methyl ester cannot be directly utilized by luciferase; it must first be hydrolyzed by esterase to produce D-luciferin, which can then be oxidized by luciferase to emit fluorescence.

<Checkpoint>[Probe $\mathbf{A}$ must undergo esterase hydrolysis before generating fluorescence](0.5)</Checkpoint>

For $\mathbf{B}$, the reaction process proposed in the article is as follows:

Probe B has a nitrobenzene-modified thiourea-based reactive group at its terminus. Studies indicate that this group can react with nitric oxide in the air to form an oxadiazole intermediate, which is then hydrolyzed to form D-aminoluciferin methyl ester. Refer to https://pubs.acs.org/doi/10.1021/acs.inorgchem.6b02787 and https://pubs.acs.org/doi/10.1021/acssensors.8b00776

<MultiModal>![The specific thiourea structure and its reaction mechanism with NO in air, as well as the kinetic studies, do indeed confirm this. ref:https://pubs.acs.org/doi/10.1021/acs.inorgchem.6b02787](/media/uploads/185948d4-0f76-4636-8d46-8ddacc235e39.png)</MultiModal>

<Checkpoint>[Probe $\mathbf{B}$ can react directly with NO](1.0)</Checkpoint>

D-aminoluciferin methyl ester also requires hydrolysis by esterase before it can be utilized by luciferase. In other words, both $\mathbf{A}$ and $\mathbf{B}$ require enzymatic processing to generate fluorescence.

<Checkpoint>[Probe $\mathbf{B}$ must undergo esterase hydrolysis before generating fluorescence](0.5)</Checkpoint>

Thus, we find that none of the options are correct, so we select option I.
研究者们开发了两种生物荧光探针（A和B），用于选择性检测生物体内的超氧化物和一氧化氮。使用的“生物荧光探针”是类似于生物体内萤火虫的发光机制，发光过程需要萤光素酶的参与。

<MultiModal>![文章提出的A、B两种探针产生荧光的机理，从上到下，从左到右对应的SMILES分别是：

O=C(N(CCN(C)C(OC1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1)=O)C)OC4=CC(OC(C)=O)=C(OC(C)=O)C=C4

OC1=C(O)C=CC(OC(N(CCN(C)C(OC2=CC(SC(C3=N[C@H](C(OC)=O)CS3)=N4)=C4C=C2)=O)C)=O)=C1

O=C1C(C=CC(OC(N(CCN(C)C(OC2=CC(SC(C3=N[C@H](C(OC)=O)CS3)=N4)=C4C=C2)=O)C)=O)=C1)=O

OC1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1

S=C(NNC(N(C1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1)C)=O)NC4=CC=C([N+]([O-])=O)C=C4

O=C(OC)[C@H](CS1)N=C1C2=NC3=C(S2)C=C(NC4=NN=C(NC5=CC=C([N+]([O-])=O)C=C5)O4)C=C3

NC1=CC(SC(C2=N[C@H](C(OC)=O)CS2)=N3)=C3C=C1](/media/uploads/4e683c29-fa56-4878-9934-cf531a51b78d.png)</MultiModal>

<Checkpoint>[产生荧光利用了荧光素系统](0.5)</Checkpoint>

分析结构可以发现，两种探针分子都含有一个共同的核心骨架，其SMILES式为 `O=C(OC)[C@H](CS1)N=C1C2=NC(C=C3)=C(S2)C=C3OC`。这是一个D-萤光素甲酯的衍生物。其中D-萤光素是萤火虫发光体系中的底物，它在ATP和萤光素酶的催化下氧化产生荧光。

<MultiModal>![荧光素甲酯基团的SMILES: O=C(OC)[C@H](CS1)N=C1C2=NC(C=C3)=C(S2)C=C3OC](/media/uploads/ef9f7eda-5160-4ee0-93f0-83c77f770b2c.png)</MultiModal>

<Checkpoint>[两种探针都拥有荧光素骨架](0.5)</Checkpoint>

对于$\mathbf{A}$来说，文章提出的反应过程如下：

第一步：去保护 。乙酰基通常会被生物体内的酯酶水解，生成相应的酚羟基。这是一个酶促反应。

<Checkpoint>[探针$\mathbf{A}$首先需要被水解酶水解](0.5)</Checkpoint>

第二步：与超氧化物反应 。酯酶水解后，生成邻苯二酚结构。这样的结构先被氧化为半醌，再被氧化为邻苯醌。虽然和旁边的体系形成了一定的共轭结构，但是这并不能发出荧光。

这一步的选择性可能是由于邻苯酚-半醌体系较强的还原性产生的，并且分子中其他位点的氧化可能实际上并不会影响最终的结果。

<Checkpoint>[探针$\mathbf{A}$经过邻苯醌的中间体](0.5)</Checkpoint>

第三步：水解。邻苯醌是一个缺电子的体系，它被迅速的水解，接下来被迅速水解的还有右侧荧光素甲酯衍生物的酚酯，留下来荧光素甲酯部分。

第四步：酯酶的水解。荧光素甲酯不能直接被荧光素酶利用，而是必须被酯酶水解产生荧光素，然后才能被荧光素酶氧化后发出荧光。

<Checkpoint>[探针$\mathbf{A}$在产生荧光之前必须经过酯酶水解](0.5)</Checkpoint>

对于$\mathbf{B}$来说，文章提出的反应过程如下：

探针B的末端有一个硝基苯修饰的，基于硫代氨脲的反应基团。

17年的研究给出了具体的反应机理，18年的研究使用的类似的底物，可以参考：https://pubs.acs.org/doi/10.1021/acs.inorgchem.6b02787 和 https://pubs.acs.org/doi/10.1021/acssensors.8b00776

<MultiModal>![特异的硫代氨脲结构和空气中NO反应的机理，动力学研究确实如此，ref:https://pubs.acs.org/doi/10.1021/acs.inorgchem.6b02787](/media/uploads/185948d4-0f76-4636-8d46-8ddacc235e39.png)</MultiModal>

指出这一部分可以和空气中的一氧化氮反应，形成一个噁二唑的中间体，然后被水解形成D-氨代荧光素甲酯。

<Checkpoint>[探针$\mathbf{B}$可以直接和NO反应](1.0)</Checkpoint>

D-氨代荧光素甲酯也需要被酯酶水解之后才能被荧光素酶利用。也就是说，$\mathbf{A，B}$都需要酶的处理之后才能产生荧光。

<Checkpoint>[探针$\mathbf{B}$在产生荧光之前必须经过酯酶水解](0.5)</Checkpoint>

于是我们发现选项均不正确，因此选择选项I。