Magnetic Tunability via Control of Crystallinity and Size in Polycrystalline Iron Oxide Nanoparticles.

Iron oxide nanoparticles (IONPs) are widely used for biomedical applications due to their unique magnetic properties and biocompatibility. However, the controlled synthesis of IONPs with tunable particle sizes and crystallite/grain sizes to achieve desired magnetic functionalities across single-domain and multi-domain size ranges remains an important challenge. Here, a facile synthetic method is used to produce iron oxide nanospheres (IONSs) with controllable size and crystallinity for magnetic tunability. First, highly crystalline Fe3O4 IONSs (crystallite sizes above 24 nm) having an average diameter of 50 to 400 nm are synthesized with enhanced ferrimagnetic properties. The magnetic properties of these highly crystalline IONSs are comparable to those of their nanocube counterparts, which typically possess superior magnetic properties. Second, the crystallite size can be widely tuned from 37 to 10 nm while maintaining the overall particle diameter, thereby allowing precise manipulation from the ferrimagnetic to the superparamagnetic state. In addition, demonstrations of reaction scale-up and the proposed growth mechanism of the IONSs are presented. This study highlights the pivotal role of crystal size in controlling the magnetic properties of IONSs and offers a viable means to produce IONSs with magnetic properties desirable for wider applications in sensors, electronics, energy, environmental remediation, and biomedicine.

Allosteric effects of EF-G domain I mutations inducing ribosome frameshifting revealed by multiplexed force spectroscopy.

Ribosome translocation catalyzed by elongation factor G (EF-G) is a critical step in protein synthesis where the ribosome typically moves along the mRNA by three nucleotides at each step. To investigate the mechanism of EF-G catalysis, it is essential to precisely resolve the ribosome motion at both ends of the mRNA, which, to our best knowledge, is only achieved with the magnetic-based force spectroscopy developed by our groups. Here, we introduce a novel multiplexed force spectroscopy technique that, for the first time, offers single-nucleotide resolution for multiple samples. This technique combines multiple acoustic force generators with the smallest atomic magnetometer designed for biological research. Utilizing this technique, we demonstrate that mutating EF-G at the GTP binding pocket results in the ribosome moving only two nucleotides on both ends of the mRNA, thereby compromising ribosome translocation. This finding suggests a direct link between GTP hydrolysis and ribosome translocation. Our results not only provide mechanistic insights into the role of GTP binding pocket but also illuminate how allosteric mutations can manipulate translocation. We anticipate broader applications of our technique in the ribosome field, leveraging its high efficiency and single-nucleotide resolution.

Twice exploration of tRNA +1 frameshifting in an elongation cycle of protein synthesis.

Inducing tRNA +1 frameshifting to read a quadruplet codon has the potential to incorporate a non-natural amino acid into the polypeptide chain. While this strategy is being considered for genome expansion in biotechnology and bioengineering endeavors, a major limitation is a lack of understanding of where the shift occurs in an elongation cycle of protein synthesis. Here, we use the high-efficiency +1-frameshifting SufB2 tRNA, containing an extra nucleotide in the anticodon loop, to address this question. Physical and kinetic measurements of the ribosome reading frame of SufB2 identify twice exploration of +1 frameshifting in one elongation cycle, with the major fraction making the shift during translocation from the aminoacyl-tRNA binding (A) site to the peptidyl-tRNA binding (P) site and the remaining fraction making the shift within the P site upon occupancy of the A site in the +1-frame. We demonstrate that the twice exploration of +1 frameshifting occurs during active protein synthesis and that each exploration is consistent with ribosomal conformational dynamics that permits changes of the reading frame. This work indicates that the ribosome itself is a determinant of changes of the reading frame and reveals a mechanistic parallel of +1 frameshifting with -1 frameshifting.

Infantile Cranial Fasciitis: A Clinicopathologic Evaluation.

OBJECTIVE: To investigate the clinicopathologic features, immunophenotype, molecular genetic changes, and differential diagnosis of cranial fasciitis (CF). METHODS: The clinical manifestations, imaging, surgical technique, pathologic characteristics, special staining, and immunophenotype, as well as break-apart fluorescence in situ hybridization assay for USP6 of 19 CF cases were analyzed, retrospectively. RESULTS: The patients were 11 boys and 8 girls, aged 5 to 144 months, with a median age of 29 months. There were 5 cases (26.31%) in the temporal bone, 4 cases (21.05%) in the parietal bone, 3 cases (15.78%) in the occipital bone, 3 cases (15.78%) in the frontotemporal bone, 2 cases (10.52%) in the frontal bone, 1 case (5.26%) in the mastoid of middle ear, and 1 case (5.26%) in the external auditory canal. The main clinical manifestations were painless, with the presentation of masses that grew rapidly and frequently eroded the skull. There was no recurrence and no metastasis after the operation. Histologically, the lesion consists of spindle fibroblasts/myofibroblasts arranged in bundles, braided or atypical spokes. Mitotic figures could be seen, but not atypical forms. Immunohistochemical studies showed diffuse strong positive SMA and Vimentin in all CFs. These cells were negative for Calponin, Desmin, ß-catenin, S-100, and CD34. The ki-67 proliferation index was 5% to 10%. Ocin blue-PH2.5 staining showed blue-stained mucinous features in the stroma. The positive rate of USP6 gene rearrangement detected by fluorescence in situ hybridization assay was about 10.52%, and the positive rate was not related to age. All patients were observed for 2 to 124 months and showed no signs of recurrence or metastasis. CONCLUSIONS: In summary, CF was a benign pseudosarcomatous fasciitis that occurs in the skull of infants. Preoperative diagnosis and differential diagnosis were difficult. Computed tomography typing might be beneficial for imaging diagnosis, and pathologic examination might be the most reliable way to diagnose CF.

Force-Induced Visualization of Nucleic Acid Functions with Single-Nucleotide Resolution.

Nucleic acids are major targets for molecular sensing because of their wide involvement in biological functions. Determining their presence, movement, and binding specificity is thus well pursued. However, many current techniques are usually sophisticated, expensive, and often lack single-nucleotide resolution. In this paper, we report the force-induced visualization method that relies on the novel concept of mechanical force to determine the functional positions of nucleic acids with single-nucleotide resolution. The use of an adjustable mechanical force overcomes the variation of analyte concentration and differences in buffer conditions that are common in biological settings. Two examples are described to validate the method: one is probing the mRNA movement during ribosomal translocation, and the other is revealing the interacting sites and strengths of DNA-binding drugs based on the force amplitude. The flexibility of the method, simplicity of the associated device, and capability of multiplexed detection will potentially enable a broad range of biomedical applications.

Clinicopathological Analysis of Sturge-Weber Syndrome with Focal Cortical Dysplasia FCD IIIc.

Objective: To investigate the clinicopathological features of children with Sturge-Weber syndrome and to analyze the correlation between the distribution area of leptomeningeal angiomatosis, the degree of cerebral cortical calcification, and the degree of cerebral atrophy associated with epileptic seizures. Methods: 10 children were diagnosed with SWS with FCD IIIc by histopathology and immunohistochemistry. Spearman correlation analysis was used to calculate the association of SWS with FCD IIIc and seizures in children. Results: The leptomeningeal angiomatosis area was markedly positively correlated with the degree of brain atrophy in 10 children with SWS (r = 0.783, p = 0.007). The distribution of leptomeningeal hemangiomatosis, the degree of cortical calcification, and brain atrophy were not significantly correlated with epilepsy. Conclusion: SWS may be accompanied by FCD IIIc. The more extensive the cerebral lobes of leptomeningeal angiomatosis in SWS, the more pronounced the brain atrophy.

High-Resolution DNA Dual-Rulers Reveal a New Intermediate State in Ribosomal Frameshifting.

Ribosomal frameshifting is an important pathway used by many viruses for protein synthesis that involves mRNA translocation of various numbers of nucleotides. Resolving the mRNA positions with subnucleotide precision will provide critical mechanistic information that is difficult to obtain with current techniques. We report a method of high-resolution DNA rulers with subnucleotide precision and the discovery of new frameshifting intermediate states on mRNA containing a GA7 G motif. Two intermediate states were observed with the aid of fusidic acid, one at the "0" reading frame and the other near the "-1" reading frame, in contrast to the "-2" and "-1" frameshifting products found in the absence of the antibiotic. We termed the new near-"-1" intermediate the Post(-1*) state because it was shifted by approximately half a nucleotide compared to the normal "-1" reading frame at the 5'-end. This indicates a ribosome conformation that is different from the conventional model of three reading frames. Our work reveals uniquely precise mRNA motions and subtle conformational changes that will complement structural and fluorescence studies.

Abnormal white matter within brain structural networks is associated with high-impulse behaviour in codeine-containing cough syrup dependent users.

Codeine-containing cough syrup (CCS) is considered as one of the most popular drug of dependence among adolescents because of its inexpensiveness and easy availability. However, its relationship with neurobiological effects remains sparsely explored. Herein, we examined how high-impulse behaviours relate to changes in the brain structural networks. Forty codeine-containing cough syrup dependent (CCSD) users and age-, gender-, and number of cigarettes smoked per day -matched forty healthy control (HC) subjects underwent structural brain imaging via MRI. High-impulse behaviour was assessed using the 30-item self-rated Barratt Impulsiveness Scale (BIS-11), and structural networks were constructed using diffusion tensor imaging and AAL-90 template. Between-group topological metrics were compared using nonparametric permutations. Benjamin-Hochberg false discovery rate correction was used to correct for multiple comparisons (P < 0.05). The relationships between abnormal network metrics and clinical characteristics of CCS dependent (BIS-11 total score, CCS- dependent duration and mean dose) were examined by Spearman's correlation. Structural networks of the CCSD group demonstrated lower small-world properties than those of the HC group. Abnormal changes in nodal properties among CCSD users were located mainly in the frontal gyrus, inferior parietal lobe and olfactory cortex. NBS analysis further indicated disrupted structural connections between the frontal gyrus and multiple brain regions. There were significant correlations between abnormal nodal properties of the frontal gyrus and clinical characteristics (BIS-11 total score, CCS dependent duration and mean dose) in the CCSD group. These findings suggest that the high-impulse behavioural expression in CCS addiction is associated with widespread brain regions, particularly within those in the frontal cortex. Aberrant brain regions and disrupted connectivity of structural network may be the bases of neuropathology for underlying symptoms of high-impulse behaviours in CCSD users, which may provide a novel sight to better treat and prevent codeine dependency in adolescents.

Ionomics analysis provides new insights into the co-enrichment of cadmium and zinc in wheat grains.

Cadmium (Cd) is present in many soils and, when enter a food chain, represents a major health threat to humans. The existent large variation in grain Cd content amongst wheat genotypes opens prospects for genetic improvement for reduced Cd uptake in this species. However, selecting low-Cd-accumulating varieties comes with a possible caveat of affecting uptake other essential nutrients. In this work, we screened 134 wheat varieties in 3 various field studies and selected 15 high- and 15 low-Cd accumulating varieties in grains for ionomics analysis. Our results showed that high-Cd accumulating varieties also possessed an ability to accumulate mineral elements of calcium, magnesium, manganese, iron and zinc, while varieties with low Cd content were deficient in many essential nutrients and, especially, zinc (Zn). The above data was confirmed in an independent trail involving another 97 wheat varieties. Thus, selecting plants for high Zn accumulation (as a part of biofortification programs) resulted in an inadvertent increase in accumulation of the toxic Cd in wheat. Vice versa, selecting low Cd-accumulating varieties comes with a danger of reducing their Zn content, with major consequences to food quality and human health. We suggest that the above conundrum can be resolved by understanding the structure-function relations of various transporters isoforms involved in Zn and Cd transport and issue-specific mode of their operation, via cell-based phenotyping followed by molecular breeding.

Antimony symplastic and apoplastic absorption, compartmentation, and xylem translocation in Brassica parachinensis L. under antimonate and antimonite.

Antimony (Sb) excess accumulation in edible parts of crops causes potential risks to human health. However, knowledge about the mechanisms of its accumulation within vegetable plants is still not well known. Here, we investigated the physiological processes of Sb involved in symplastic and apoplastic absorption, compartmentation by roots, and translocation in xylem in Brassica parachinensis L. exposed to antimonate (SbV) and antimonite (SbIII) forms. The results showed that plants treated with SbIII emerged to be more toxic than SbV as proved by the lower biomass and the higher concentrations of malonaldehyde (MDA) and hydrogen peroxide (H2O2) in plant tissues, especially at high dosages. The Sb concentration showed more in shoots but less in roots treated with SbV than with SbIII. The total Sb accumulation was higher under the SbV treatment than the SbIII treatment, mainly due to the higher accumulation in shoots. Additionally, the Sb concentration in symplastic flow of roots was higher exposed to SbV than SbIII, while no differences were found for the Sb concentration in apoplastic flow between them. Moreover, the Sb concentration in cell walls of roots was higher exposed to SbIII than SbV, especially at high levels. Furthermore, the Sb concentration in xylem was higher exposed to SbV than SbIII, and a greatly positive correlation was observed between the Sb concentrations in xylem and shoots. Overall, these findings revealed that vegetable plants accumulated more SbV than SbIII in edible parts mainly due to xylem translocation rather than root absorption.

Ammonium nutrition mitigates cadmium toxicity in rice (Oryza sativa L.) through improving antioxidase system and the glutathione-ascorbate cycle efficiency.

Nitrogen (N) forms not only affect cadmium (Cd) accumulation in plants, but also affect plant resistance to Cd toxicity. However, few researches have been reported underlying the mechanism of the relationship between nitrogen forms and plant resistance under Cd exposure. Here, we explored the mechanism on how different NO3-/NH4+ ratios affect antioxidase system and the glutathione-ascorbate cycle under five different ratios of NO3-/NH4+ (1:0, 2:1, 1:1, 1:2, 0:1) and three dosages of Cd exposure (0, 1, 5 µmol L-1 Cd) in rice (Oryza sativa L.). The results showed that high NO3- and high Cd exposure both significantly inhibited tissue growth of rice plants, and this inhibiting trend was mitigated with increasing NH4+ ratios as proved by the increased biomass and the decreased concentrations of malonaldehyde (MDA) and hydrogen peroxide (H2O2), as well as the levels of Cd contents in rice tissues. Additionally, high NH4+ ratios elevated the SOD activities in rice tissues, especially at high Cd treatment. However, other two antioxidases (CAT and APX) were insensitive to changes of NO3-/NH4+ ratios (except the full NO3-). Furthermore, high NH4+ ratios induced increasing of the efficiency of glutathione-ascorbate cycle (GSH-AsA) under two levels of Cd exposure, as evidenced by increasing concentrations of GSH and AsA and the activities of GR and DHAR in rice tissues. Overall, these results revealed that ammonium nutrition caused an enhancement resistance to Cd stress in rice plants was responsible for increasing of partial antioxidase system and the efficiencies of GSH-AsA cycle.

Modulation and Visualization of EF-G Power Stroke During Ribosomal Translocation.

During ribosome translocation, the elongation factor EF-G undergoes large conformational change while maintaining its contact with the moving tRNA. We previously measured a power stroke accompanying EF-G catalysis, which was consistent with structural studies. However, the role of power stroke in translocation fidelity remains unclear. Here, we report quantitative measurements of the power strokes of structurally modified EF-Gs by using two different techniques and reveal the correlation between power stroke and translocation efficiency and fidelity. We discovered that the reduced power stroke only lowered the percentage of translocation but did not introduce translocation error. The established force -structure-function correlation for EF-G indicates that power stroke drives ribosomal translocation, but the mRNA reading frame is probably maintained by ribosome itself. Furthermore, the microscope detection method reported here can be simply implemented for other biochemical applications.

Specific Detection of Proteins Using Exceptionally Responsive Magnetic Particles.

Sensitivity and specificity are among the most important parameters for viable sensor technologies based on magnetic nanoparticles. In this work, we describe synthetic routes and analytical approaches to improve both aspects. Magnetic iron oxide particles having diameters of 120, 440, and 700 nm were synthesized, and their surfaces were specifically functionalized. The larger particles showed significantly stronger magnetic signals and responses when compared to commercially available magnetic particles (Dynabeads). A force-based detection method was used to distinguish specifically bound particles (via protein interactions) and nonspecifically bound ones (e.g., via physisorption). In addition, an exchange platform, denoted as exchange-induced remnant magnetization (EXIRM), was developed and utilized to detect label-free proteins specifically. Using EXIRM, the 700 nm magnetic particles showed a 7-fold increase in detection sensitivity when compared to the markedly larger commercially available Dynabeads; furthermore, EXIRM exhibited high specificity, even in a 100-fold increase of nontargeted protein. More generally, particle size effects, reaction times, and dynamic ranges are evaluated and discussed herein.

Dual DNA rulers reveal an 'mRNA looping' intermediate state during ribosome translocation.

The precise 3-nucleotide movement of mRNA is critical for translation fidelity. One mRNA translocation error propagates to all of the following codons, which is detrimental to the cell. However, none of the current methods can reveal the mRNA dynamics near the ribosome entry site, which limits the understanding of this important issue. We have developed an assay of dual DNA rulers that provides such capability. By uniquely probing both the 3'- and 5'-ends of mRNA, we observed an antibiotic-trapped intermediate state that is consistent with a ribosomal conformation containing mRNA asymmetric partial displacements at its entry and exit sites. Based on the available ribosome structures and computational simulations, we proposed a 'looped' mRNA conformation, which suggested a stepwise 'inchworm' mechanism for ribosomal translocation. The same 'looped' intermediate state identified with the dual rulers persists with a '-1' frameshifting motif, indicating that the branching point of normal and frameshifting translocations occurs at a later stage of translocation.

Nitric oxide acts downstream of abscisic acid in molybdenum-induced oxidative tolerance in wheat.

KEY MESSAGE: Our study first reveals that Mo mediates oxidative tolerance through ABA signaling. Moreover, NO acts downstream of ABA signaling in Mo-induced oxidative tolerance in wheat under drought stress. Nitric oxide (NO) is related to the improvement of molybdenum (Mo)-induced oxidative tolerance. While the function of Mo in abscisic acid (ABA) synthesis and in mediating oxidative tolerance by the interaction of ABA and NO remain to be studied. The -Mo and +Mo treatment-cultivated wheat was separated and subsequently was pretreated with AO inhibitor, ABA synthesis inhibitor, exogenous ABA, NO scavenger, NO donor or their combinations under polyethylene glycol 6000 (PEG)-stimulated drought stress (PSD). The AO activity and ABA content were increased by Mo in wheat under PSD, however, AO inhibitor decreased AO activity, correspondingly reduced ABA accumulation, suggesting that AO involves in the regulation of Mo-induced ABA synthesis. Mo enhanced activities and expressions of antioxidant enzyme, while these effects of Mo were reversed by AO inhibitor and ABA synthesis inhibitor due to the decrease of ABA content, but regained by exogenous ABA, indicating that Mo induces oxidative tolerance through ABA. Moreover, NO scavenger inhibited activities of antioxidant enzyme caused by Mo and exogenous ABA, but the inhibitions were eliminated by NO donor, indicating that NO is involved in ABA pathway in the regulation of Mo-induced oxidative tolerance in wheat under PSD. Finally, we proposed a scheme for the mechanism of Mo-induced oxidative tolerance.

Metabolomics analysis reveals potential mechanisms of tolerance to excess molybdenum in soybean seedlings.

Most plants exhibit strong tolerance to excess molybdenum (Mo). However, the metabolic profile and tolerance mechanisms of plants in response to excess Mo remain unknown. We comprehensively analyzed changes in the metabolic profiles of leaves and roots in soybean (Glycine max L.) seedlings cultured under normal-Mo and excess-Mo conditions by using ultra performance liquid chromatography (UPLC) combined with MS/MS (mass spectrometry). There were 42 differential metabolites in the roots and 19 differential metabolites in the leaves in response to excess Mo stress. In roots, the organic acids, levels of gluconic acid, D-glucarate and citric acid increased by 107.63-, 4.42- and 2.87-folds after excess Mo exposure. Several hormones (salicylic acid, jasmonic acid) and lipids (PG, MG, DG etc) also increased significantly under excess Mo condition. Metabolites related to ascorbate-glutathione metabolism and flavonoid and isoflavone biosynthesis notably accumulated in roots. Only lipid metabolism and salicylic acid accumulation were induced in leaves under excess Mo stress. It is speculated that organic compounds such as 2-oxoarginine, L-nicotine, gluconic acid, D-glucurate, and citric acid played important roles to chelate Mo and reduce its toxicity. Signaling molecules (JA, SA, and some lipids) and non-enzyme antioxidants such as flavonoids/isoflavones act synergistically to detoxify ROS and contribute to Mo tolerance in soybean seedlings. More metabolic pathways were induced by Mo excess in roots than in leaves, suggesting that roots play more implant role in Mo tolerance.

Comparison of foliar silicon and selenium on cadmium absorption, compartmentation, translocation and the antioxidant system in Chinese flowering cabbage.

Silicon (Si) and selenium (Se) are beneficial for many higher plants when grown on stress conditions. However, the mechanisms underlying the differential effects between foliar Si and Se in alleviation of plant toxicity exposed to cadmium (Cd) stress are remained unclear. In this study, we investigated the discrepant mechanisms of foliar Si and Se on Cd absorption and compartmentation by roots, its translocation in xylem, and the antioxidant system within Chinese flowering cabbage (Brassica campestris L. ssp. chinensis var. utilis) under low and high Cd stress. Results showed that plant growth was significantly enhanced by foliar additions of Si or/and Se according to an increased plant tissue biomass at high Cd exposure. In addition, the foliar coupled addition of Si and Se showed little effects on the concentrations of Si or Se in plant tissues in comparison with the single addition of foliar Si or Se respectively. The foliar Si alone or combined with Se markedly reduced the Cd concentrations in plant shoots under two Cd treatments. This might be explained by the lower Cd concentrations in symplast and apoplast and the higher Cd concentrations in cell walls of plant roots, and the lower Cd concentrations in xylem sap. However, no great changes in these values were observed under the treatments of foliar Se alone. Moreover, the foliar additions of Si or/and Se all increased the antioxidant enzyme activities of SOD, CAT and APX in plant tissues, especially at high Cd dosage. No significant differences in the increasing degrees of these three antioxidant enzymes were found between the foliar Si and Se treatments. However, only the foliar Se alone or combined with Si markedly promoted the antioxidant enzyme activities of GR and DHAR in plant tissues. Our findings demonstrate that the alleviation of Cd toxicity by foliar Si maybe mainly responsible for inhibition of Cd absorption and its translocation to plant shoots, reinforcing its compartmentation into root cell walls, whilst enhancing the antioxidant enzyme system may be employed by foliar Se.

Biosensing Using Magnetic Particle Detection Techniques.

Magnetic particles are widely used as signal labels in a variety of biological sensing applications, such as molecular detection and related strategies that rely on ligand-receptor binding. In this review, we explore the fundamental concepts involved in designing magnetic particles for biosensing applications and the techniques used to detect them. First, we briefly describe the magnetic properties that are important for bio-sensing applications and highlight the associated key parameters (such as the starting materials, size, functionalization methods, and bio-conjugation strategies). Subsequently, we focus on magnetic sensing applications that utilize several types of magnetic detection techniques: spintronic sensors, nuclear magnetic resonance (NMR) sensors, superconducting quantum interference devices (SQUIDs), sensors based on the atomic magnetometer (AM), and others. From the studies reported, we note that the size of the MPs is one of the most important factors in choosing a sensing technique.

The onset of electron-induced proton-transfer in hydrated azabenzene cluster anions.

The prospect that protons from water may be transferred to N-heterocyclic molecules due to the presence of an excess electron is studied in hydrated azabenzene cluster anions using anion photoelectron spectroscopy and computational chemistry. In the case of s-triazine (C3H3N3), which has a positive adiabatic electron affinity, proton transfer is not energetically favored nor observed experimentally. Heterocyclic rings with only 1 or 2 nitrogen atoms have negative electron affinities, but the addition of solvating water molecules can yield stable negative ions. In the case of the diazines (C4H4N2: pyrazine, pyrimidine, and pyridazine) the addition of one water molecule is enough to stabilize the negative ion, with the majority of the excess electron density in a π* orbital of the heterocycle but not significantly extended over the hydrogen bonded water network. Pyridine (C5H5N), with the most negative electron affinity, requires three water molecules to stabilize its negative ion. Although our computations suggest proton transfer to be energetically viable in all five N-heterocyclic systems studied here when three or more water molecules are present, proton transfer is not observed experimentally in the triazine nor in the diazine series. In pyridine, however, proton transfer competes energetically with hydrogen bonding (solvation), when three water molecules are present, i.e., both motifs are observed. Pyridine clusters containing four or more water molecules almost exclusively exhibit proton transfer along with solvated [C(6-x)H(6-x+1)N(x)·OH](-) ions.

Photoelectron spectroscopic and computational study of hydrated pyrimidine anions.

The stabilization of the pyrimidine anion by the addition of water molecules is studied experimentally using photoelectron spectroscopy of mass-selected hydrated pyrimidine clusters and computationally using quantum-mechanical electronic structure theory. Although the pyrimidine molecular anion is not observed experimentally, the addition of a single water molecule is sufficient to impart a positive electron affinity. The sequential hydration data have been used to extrapolate to -0.22 eV for the electron affinity of neutral pyrimidine, which agrees very well with previous observations. These results for pyrimidine are consistent with previous studies of the hydrated cluster anions of uridine, cytidine, thymine, adenine, uracil, and naphthalene. This commonality suggests a universal effect of sequential hydration on the electron affinity of similar molecules.