Identification of reaction sites and chlorinated products of purine bases and nucleosides during chlorination: a computational study.

Hypochlorous acid (HOCl) released from activated leukocytes plays a significant role in the human immune system, but is also implicated in numerous diseases due to its inappropriate production. Chlorinated nucleobases induce genetic changes that potentially enable and stimulate carcinogenesis, and thus have attracted considerable attention. However, their multiple halogenation sites pose challenges to identify them. As a good complement to experiments, quantum chemical computation was used to uncover chlorination sites and chlorinated products in this study. The results indicate that anion salt forms of all purine compounds play significant roles in chlorination except for adenosine. The kinetic reactivity order of all reaction sites in terms of the estimated apparent rate constant kobs-est (in M-1 s-1) is heterocyclic NH/N (102-107) > exocyclic NH2 (10-2-10) > heterocyclic C8 (10-5-10-1), but the order is reversed for thermodynamics. Combining kinetics and thermodynamics, the numerical simulation results show that N9 is the most reactive site for purine bases to form the main initial chlorinated product, while for purine nucleosides N1 and exocyclic N2/N6 are the most reactive sites to produce the main products controlled by kinetics and thermodynamics, respectively, and C8 is a possible site to generate the minor product. The formation mechanisms of biomarker 8-Cl- and 8-oxo-purine derivatives were also investigated. Additionally, the structure-kinetic reactivity relationship study reveals a good correlation between lg kobs-est and APT charge in all purine compounds compared to FED2 (HOMO), which proves again that the electrostatic interaction plays a key role. The results are helpful to further understand the reactivity of various reaction sites in aromatic compounds during chlorination.

Insights into C-C Bond Cleavage Mechanisms in Dichloroacetonitrile Formation during Chlorination of Long-Chain Primary Amines, Amino Acids, and Dipeptides.

Dichloroacetonitrile (DCAN) as one of the potentially prioritized regulated DBPs has drawn great attention; however, understanding its formation, especially the C-C bond cleavage mechanisms, is limited. In this study, DCAN formation mechanisms from long-chain primary amines, amino acids, and dipeptides during chlorination were investigated by a combined computational and experimental approach. The results indicate that nitriles initially generate for all of the above precursors, then they undergo ß-C-hydroxylation or/and α-C-chlorination processes, and finally, DCAN is produced through the Cα-Cß bond cleavage. For the first time, the underlying mechanism of the C-C bond cleavage was unraveled to be electron transfer from the O- anion into its attached C atom in the chlorinated nitriles, leading to the strongly polarized Cα-Cß bond heterocleavage and DCAN- formation. Moreover, DCAN molar yields of precursors studied in the present work were found to be determined by their groups at the γ-site of the amino group, where the carbonyl group including -CO2-, -COR, and -CONHR, the aromatic group, and the -OH group can all dramatically facilitate DCAN formation by skipping over or promoting the time-consuming ß-C-hydroxylation process and featuring relatively lower activation free energies in the C-C bond cleavage. Importantly, 4-amino-2-hydroxybutyric acid was revealed to possess the highest DCAN yield among all the known aliphatic long-chain precursors to date during chlorination. Additionally, enonitriles, (chloro-)isocyanates, and nitriles can be generated during DCAN formation and should be of concern due to their high toxicities.

Computational Investigations of Reaction Mechanisms and Transformation Products of Olefins with Hypochlorous Acid.

Hypochlorous acid (HOCl) as the main component in chlorination and also as the innate immune factor relevant to immune defense has attracted considerable attention. Electrophilic addition reaction of olefins with HOCl, one of the most important prototype of chemical reactions, has been intensively studied for a long time; however, it has not been fully understood yet. In this study, addition reaction mechanisms and transformation products of model olefins with HOCl were systematically investigated by the density functional theory method. The results indicate that the traditionally believed stepwise mechanism with a chloronium-ion intermediate is only suitable for olefins substituted with electron-donating groups (EDGs) and weak electron-withdrawing groups (EWGs) but it is a carbon-cation intermediate that is favorable for EDGs featuring p-π or π-π conjugation with the C═C moiety. Moreover, olefins substituted with moderate or/and strong EWGs prefer the concerted and nucleophilic addition mechanisms, respectively. Epoxide and truncated aldehyde as the main transformation products can be generated from chlorohydrin through a series of reactions involving hypochlorite; however, their generation is kinetically not as feasible as the formation of chlorohydrin. The reactivity of three chlorinating agents (HOCl, Cl2O, and Cl2) and the case study of chlorination and degradation of cinnamic acid were also explored. Additionally, APT charge on the double-bond moiety in olefin and energy gap (ΔE) between the highest occupied molecular orbital (HOMO) energy of olefin and the lowest unoccupied molecular orbital (LUMO) energy of HOCl were found to be good parameters to distinguish the regioselectivity of chlorohydrin and reactivity of olefin, respectively. The findings of this work are helpful in further understanding the chlorination reactions of unsaturated compounds and identifying complicated transformation products.

IRF7 suppresses hematopoietic regeneration under stress via CXCR4.

Hematopoietic stem cells (HSCs) maintain quiescence under steady state; however, they are compelled to proliferate and expand to replenish the blood system under stress. The molecular basis underlying stress hematopoiesis remains to be fully understood. In this study, we reported that IRF7 represents an important regulator of stress hematopoiesis. Interferon regulatory factor 7 (IRF7) was dispensable for normal hematopoiesis, whereas its deficiency significantly enhanced hematopoietic stem and progenitor cells (HSPCs) regeneration and improved long-term repopulation of HSCs under stress. Mechanistic studies showed that CXCR4 was identified as a downstream target of IRF7. Overexpression of CXCR4 abrogated the enhanced proliferation and regeneration of IRF7-deficient HSPCs under stress. Similar results were obtained in HSCs from human umbilical cord blood. These observations demonstrated that IRF7 plays an important role in hematopoietic regeneration under stress.

Reaction Mechanisms of Histidine and Carnosine with Hypochlorous Acid Along with Chlorination Reactivity of N-Chlorinated Intermediates: A Computational Study.

Hypochlorous acid (HOCl) released from activated leukocytes not only plays a significant role in the human immune system but is also implicated in numerous diseases including atherosclerosis and some cancers due to its inappropriate production. Histidine (His) and carnosine (Car), as a respective mediator and protective agent of HOCl damage, have attracted considerable attention; however, their detailed reaction mechanisms are still unclear. In this study, using a His residue with two peptide bond groups (HisRes) as a model, the reaction mechanisms of HisRes and Car including NεH and NδH tautomers with HOCl along with the chlorination reactivity of N-chlorinated intermediates were investigated by quantum chemical methods. The obtained results indicate that in the imidazole side chain, the pyridine-like N is the most reactive site rather than the pyrrole-like N, and the kinetic order of all of the possible reaction sites in HisRes follows pyridine-like N > imidazole Cδ ≫ imidazole Cε > pyrrole-like N, while that in Car is pyridine-like N ≫ imidazole Cδ ≫ amide N. As for N-chlorinated intermediates at imidazole, although the unprotonated form has a low chlorination reactivity as expected, it can still chlorinate tyrosine. Especially, the protonated form exhibits similar ability to HOCl, causing secondary damage in vivo. N-Chlorinated Car features higher internal chlorine migration ability than its intermolecular transchlorination, preventing further HOCl-induced damage. Additionally, a generally overlooked nucleophilic Cl- shift is also found in N-chlorinated Car/HisRes, indicating that nucleophilic sites in biomolecules also need to be considered. The outcomes of this study are expected to expand our understanding of secondary damage and protective mechanisms involved in HOCl in humans.

Cyclooxygenase-1 Regulates the Development of Follicular Th Cells via Prostaglandin E2.

Cyclooxygenase (COX)-1, one of the critical enzymes required for the conversion of arachidonic acid to PGs, has been demonstrated to play an important role not only in the cardiovascular system but also in the immune system. COX-1 has been found to regulate early B cell differentiation, germinal center formation, and Ab production of B cells. However, the underlying mechanisms of COX-1-mediated B cell activation remains not fully understood. In this study, we reported that COX-1 is a potential regulator for the development of follicular Th (TFH) cells. COX-1-deficient (COX-1-/- ) mice displayed a significant reduction of TFH cells upon influenza infection or immunization with keyhole limpet hemocyanin, which led to a severe impairment of germinal center responses. We further demonstrated that COX-1-derived PGE2, via binding with its receptors EP2/EP4, represents the underlying mechanism. The administration of EP2/EP4 agonists or PGE2 almost completely rescued the defective TFH cell generation in COX-1-/- mice. Taken together, our observations indicate that COX-1 plays an important role in the development of TFH cells.

Degradation Mechanisms and Substituent Effects of N-Chloro-α-Amino Acids: A Computational Study.

N-Chloro-α-amino acids formed in the chlorination disinfection treatment of water or wastewater and in living organisms have attracted extensive attention due to the potential toxicities of themselves and their decomposition products. The degradation mechanisms of three N-chloro-α-amino acids, i.e., N-chloro-glycine, N-chloro-alanine, and N-chloro-valine, have been systematically investigated using quantum chemical computations. The results indicate that N-chloro-α-amino acid anions undergo two competitive degradation pathways: a concerted Grob fragmentation (CGF) and ß-elimination (ß-E). Generally, the former predominates over the latter under neutral conditions and finally generates amines and carbonyls, while the latter is preferred under base-promoted conditions and mainly produces the respective α-keto acid anions or nitriles in the end. To gain deeper insights into the substitution effects, in view of the advantages of quantum chemical computations, a number of real or designed N-chloro-α-amino acids with traditional electron-donating groups (EDG) or electron-withdrawing groups (EWG) have been studied. All of the substituted N-chloro-α-amino acids, regardless of the type and position of substituents, are kinetically more favorable than N-monochloro-glycine for degradation via the CGF pathway. Moreover, conjugated EDG substituted on the N-terminal facilitate both CGF and ß-E reactions, whereas conjugated EDG and EWG on the α-carbon are only favorable for the CGF and ß-E reactions, respectively. These results are expected to expand our understanding of organic N-chloramine degradation mechanisms and chlorination reaction characteristics.

Schisandrin B ameliorates high-glucose-induced vascular endothelial cells injury by regulating the Noxa/Hsp27/NF-κB signaling pathway.

BACKGROUND: To address the molecular mechanism of the anti-inflammation effects of schisandrin B (Sch B) in atherosclerosis, we examined injured HMEC-1, HBMEC, and HUVEC-12 cells induced by high glucose (HG). METHODS: Western blot was performed to detect the levels of the proteins Hsp27, Noxa, TLR5, p-IκBα, and p-p65 in HG-induced cells, while ELISA was used to analyze the inflammatory cytokines TNF-α, IL-6, MCP-1, and IL-1ß in cells with Hsp27 or Noxa stable expression. RESULTS: Overexpression of Hsp27 upregulated the inflammatory cytokines and the release of IκBα, promoted transportation of p65 into the nucleus, and lastly, affected the inflammation process, while Sch B counteracted the upregulation. In addition, the effect of Noxa overexpression, which is different from Hsp27 overexpression, was consistent with that of Sch B treatment. CONCLUSIONS: Sch B may inhibit the inflammatory cascade and alleviate the injury to HMEC-1, HBMEC, and HUEVC-12 cells caused by HG by regulating the Noxa/Hsp27/NF-κB signaling pathway.

Theoretical Investigation of the Gas-Phase SN2 Reactions of Anionic and Neutral Nucleophiles with Chloramines.

The SN2 reactions at nitrogen center (SN2@N) play a significant role in organic synthesis, carcinogenesis, and the formation of some environmentally toxic compounds. However, the SN2@N reactions specifically for neutral compounds as nucleophiles are less known. In this work, reactions of dimethylamine (DMA) and F- with NH2Cl were investigated as model reactions to validate an accurate functional from 24 DFT functionals by comparing with the CCSD(T) reference data. M06-2X functional was found to perform best and applied to systematically explore the trends in reactivity for halides (F- and Cl-) and simple amines toward the substrates NH2Cl and NHCl2 (SN2@N) as well as CH3Cl and CH2Cl2 (SN2@C). The computational results show that the backside inversion channel dominates most the SN2@N reactions except for the case of F- + NHCl2, which reacts preferentially via proton transfer. The overall activation free energies (Δ G‡) of the inversion channel for the SN2 reactions of F- and Cl- with chloramines are negative, whereas those for amines as nucleophiles are around 30-44 kcal/mol. The SN2@N reactions for all the nucleophiles investigated here are faster than the corresponding SN2@C. Moreover, amines react faster when they have a higher extent of methyl substitution. Additionally, the energy gap between the HOMO of nucleophile and LUMO of substrate generally correlates well with Δ G‡ of the corresponding SN2 reactions, which is consistent with previous results.

Experimental and Theoretical Investigation of Effects of Ethanol and Acetic Acid on Carcinogenic NDMA Formation in Simulated Gastric Fluid.

N-nitrosodimethylamine (NDMA), as a representative of endogenously formed N-nitroso compounds (NOCs), has become the focus of considerable research interest due to its unusually high carcinogenicity. In this study, effects of ethanol and acetic acid on the formation of NDMA from dimethylamine (DMA) and nitrite in simulated gastric fluid (SGF) were investigated. Experimental results showed that ethanol in the concentrations of 1-8% (v/v) and acetic acid in the concentrations of 0.01-8% (v/v) exhibit inhibitory and promotion effects on the formation of NDMA, respectively. Moreover, they are both in a dose-dependent manner with the largest inhibition/promotion rate reaching ∼70%. Further experimental investigations indicate that ethanol and acetic acid are both able to scavenge nitrite in SGF. It implies that there are interactions of ethanol and acetic acid with nitrite or nitrite-related nitrosating agents rather than DMA. Theoretical calculations confirm the above experimental results and demonstrate that ethanol and acetic acid can both react with nitrite-related nitrosating agents to produce ethyl nitrite (EtONO) and acetyl nitrite (AcONO), respectively. Furthermore, the reactivities of ethyl nitrite, acetyl nitrite, and dinitrogen trioxide reacting with DMA were found in the order of AcONO > N2O3 ≫ EtONO. This is probably the main reason why there are completely different effects of ethanol and acetic acid on NDMA formation. On the basis of the above results, two requirements for a potential inhibitor of NOCs formation in SGF were provided. The results obtained in this study will be helpful in better understanding the inhibition/promotion mechanisms of compounds on NDMA formation in SGF and searching for protective substances to prevent carcinogenic NOCs formation.

Formation mechanism of NDMA from ranitidine, trimethylamine, and other tertiary amines during chloramination: a computational study.

Chloramination of drinking waters has been associated with N-nitrosodimethylamine (NDMA) formation as a disinfection byproduct. NDMA is classified as a probable carcinogen and thus its formation during chloramination has recently become the focus of considerable research interest. In this study, the formation mechanisms of NDMA from ranitidine and trimethylamine (TMA), as models of tertiary amines, during chloramination were investigated by using density functional theory (DFT). A new four-step formation pathway of NDMA was proposed involving nucleophilic substitution by chloramine, oxidation, and dehydration followed by nitrosation. The results suggested that nitrosation reaction is the rate-limiting step and determines the NDMA yield for tertiary amines. When 45 other tertiary amines were examined, the proposed mechanism was found to be more applicable to aromatic tertiary amines, and there may be still some additional factors or pathways that need to be considered for aliphatic tertiary amines. The heterolytic ONN(Me)2-R(+) bond dissociation energy to release NDMA and carbocation R(+) was found to be a criterion for evaluating the reactivity of aromatic tertiary amines. A structure-activity study indicates that tertiary amines with benzyl, aromatic heterocyclic ring, and diene-substituted methenyl adjacent to the DMA moiety are potentially significant NDMA precursors. The findings of this study are helpful for understanding NDMA formation mechanism and predicting NDMA yield of a precursor.

Identifying initial transformation products during chlorination of the indole moiety and unveiling their formation mechanisms.

To identify toxicity drivers within poorly characterized high-molar-weight disinfection by-products (DBPs), relatively stable high-yield initial transformation products generated from aromatic amino acids and peptides and humic substances have drawn much attention. In this study, initial transformation products in chlorination of the indole moiety in tryptophan (Trp) are proposed and their formation mechanisms were investigated using a quantum chemical computational method. The results indicate that 3-Cl-Trp+ is initially formed after the Cl+ of HOCl attacks the indole moiety, and nucleophilic addition with nucleophilic agents (H2O and OCl-) is thermodynamically preferred over deprotonation to generate 2-X-3-Cl-indoline moiety (X = OH and OCl), which is in contrast to indole. Over 25 types of initial transformation products are proposed from the 2-X-3-Cl-indoline moiety and two ring opening pathways were found at N1-C2 and C2-C3 bonds. Significantly, most structures of initial transformation products proposed based on experimental detection m/z values were confirmed using quantum chemical calculations and some new products are proposed in this work. The results are helpful to expand our understanding of the intrinsic reactivity of aromatic ring towards chlorination by hypochlorous acid.

Reaction sites of pyrimidine bases and nucleosides during chlorination: A computational study.

As important components of soluble microbial products in water, nucleobases have attracted much attention due to the high toxicity of their direct aromatic halogenated disinfection by-products (AH-DBPs) during chlorination. However, multiple halogenation sites of AH-DBPs pose challenges to identify them. In this study, reaction sites of pyrimidine bases and nucleosides during chlorination were investigated by quantum chemical computational method. The results indicate that the anion salt forms play key roles in chlorination of uracil, thymine, and their nucleosides, while neutral forms make predominant contributions to cytosine and cytidine. In view of both kinetics and thermodynamics, C5 is the most reactive site for uracil and thymine, N3/C5 and N3 for respective uridine and thymidine, N1/C5/N4 and N4 for respective cytosine and cytidine, whose estimated apparent rate constants kobs-est of ∼103, 103/102, 106/102/104, and 103 M-1 s-1, respectively, in consistent with the known experimental results. C6 in all pyrimidine compounds is hardly attacked by Cl+ in HOCl ascribed to its positive charge, but readily attacked by OH‾ in hydrolysis and the N1=C6 bond was found to possess the highest reactivity in hydrolysis among all double bonds. In addition, the structure-kinetic reactivity relationship study reveals a relatively strong correlation between lgkobs-est and APT charge in all pyrimidine compounds rather than FED2 (HOMO). The results are helpful to further understand the reactivity of various reaction sites in aromatic compounds during chlorination.

Appendicitis combined with Meckel's diverticulum obstruction, perforation, and inflammation in children: Three case reports.

BACKGROUND: Meckel's diverticulum is a common congenital malformation of the small intestine, with the three most common complications being obstruction, perforation, and inflammation. To date, only a few cases have been reported worldwide. In children, the clinical symptoms are similar to appendicitis. As most of the imaging features are nonspecific, the preoperative diagnosis is not precise. In addition, the clinical characteristics are highly similar to pediatric acute appendicitis, thus special attention is necessary to distinguish Meckel's diverticulum from pediatric appendicitis. Patients with poor disease control should undergo laparoscopic exploration to avoid serious complications, including intestinal necrosis, intestinal perforation and gastrointestinal bleeding. CASE SUMMARY: This report presents three cases of appendicitis in children combined with intestinal obstruction, which was caused by fibrous bands (ligaments) arising from the top part of Meckel's diverticulum, diverticular perforation, and diverticular inflammation. All three patients, aged 11-12 years, had acute appendicitis as their initial clinical presentation. All were treated by laparoscopic surgery with a favorable outcome. A complete dataset including clinical presentation, diagnostic imaging, surgical information, and histopathologic findings was also provided. CONCLUSION: Preoperative diagnosis of Meckel's diverticulum and its complications is challenging because its clinical signs and complications are similar to those of appendicitis in children. Laparoscopy combined with laparotomy is useful for diagnosis and treatment.

Comparison of nitrate formation mechanisms from free amino acids and amines during ozonation: a computational study.

Nitrate as a potential surrogate parameter for abatement of micropollutants, oxidant exposure, and characterizing oxidant-reactive DON during ozonation has attracted extensive attention, however, understanding of its formation mechanisms is still limited. In this study, nitrate formation mechanisms from amino acids (AAs) and amines during ozonation were investigated by the DFT method. The results indicate that N-ozonation initially occurs to produce competitive nitroso- and N,N-dihydroxy intermediates, and the former is preferred for both AAs and primary amines. Then, oxime and nitroalkane are generated during further ozonation, which are the important last intermediate products for nitrate formation from the respective AAs and amines. Moreover, the ozonation of the above important intermediates is the nitrate yield-controlling step, where the relatively higher reactivity of the CN moiety in the oxime compared to the general Cα atom in the nitroalkane explains why the nitrate yields of most AAs are higher than those from general amines, and it is the larger number of released Cα- anions, which are the real reaction sites attacked by ozone, that leads to the higher nitrate yield for nitroalkane with an electron-withdrawing group bound to the Cα atom. The good relationship between nitrate yields and activation free energies of the rate-limiting step (ΔG≠rls) and nitrate yield-controlling step (ΔG≠nycs) for the respective AAs and amines verifies the reliability of the proposed mechanisms. Additionally, the bond dissociation energy of Cα-H in the nitroalkanes formed from amines was found to be a good parameter to evaluate the reactivity of the amines. The findings here are helpful for further understanding nitrate formation mechanisms and predicting nitrate precursors during ozonation.

Reactions of amine and peroxynitrite: evidence for hydroxylation as predominant reaction and new insight into the modulation of CO2.

Peroxynitrite is related to numerous diseases including cardiovascular diseases, inflammation, and cancer. In order to expand the understanding for the toxicology of peroxynitrite in biological system, the reactions of amine (morpholine as a probe) with peroxynitrite and the modulation of CO2 were investigated by using DFT methods. The results strongly indicate that the hydroxylation of amine by peroxynitrous acid ONOOH, which was previously overlooked by most studies, is predominant relative to the widely reported nitration and nitrosation in the absence of CO2. The product N-hydroxylamine is proposed to be mainly generated via nonradical pathway (two-electron oxidation). The modulation of CO2 exhibits two main functions: (1) inhibition of hydroxylation due to the promoted consumption of peroxynitrite via fast reaction of CO2 with ONOO¯ to form ONOOCO2¯; (2) dual effect (catalysis and inhibition) of CO2 toward nitration and nitrosation. As a new insight, amine does react with CO2 and produce inert amine carbamate R2NCOO¯. This reaction has the potential to compete with the reaction of CO2 and ONOO¯, which leads to inhibition of nitration and nitrosation. The concentration of CO2 could be a critical factor determining the final effect, catalysis or inhibition. As a new finding, HCO3¯ is probably an effective catalyst for the reaction of amine and CO2. Moreover, further studies on how the different types of the amine might affect the outcome of the reactions would be an interesting topic.

Transformation mechanisms of acetaldehyde and its substituted aldehydes into the corresponding nitriles and (N-chloro)amides during chloramination: A computational study.

As an alternative disinfectant to free chlorine, monochloramine reduces the formation of regulated disinfection byproducts (DBPs); however, it also contributes to the formation of highly toxic nitrogenous DBPs (N-DBPs), especially through the aldehyde pathway. The current understanding of aldehyde pathway mechanisms is limited. In this study, the transformation pathways of acetaldehyde and its substituted aldehydes into the corresponding nitriles and (N-chloro)amides during chloramination were investigated using quantum chemical calculations. Consistent with previous studies, 1-chloroamino alcohol first forms in the chloramination of aldehydes and then undergoes competitive dehydration and HCl elimination branch reactions to generate the nitrile and (N-chloro)amide, respectively. Iminol was found to be a key intermediate for (N-chloro)amide formation. Moreover, the results indicated that acetaldehydes substituted with electron-donating groups (EDGs) and electron-withdrawing groups (EWGs) are beneficial to the formation of the respective nitriles and N-chloro-amides, while those substituted with conjugated groups (CGs) are favourable for both. Based upon the above results, in addition to acetaldehyde, other aldehydes, such as propionaldehyde, glycolaldehyde, 3-butenal, and phenylacetaldehyde, which are the α-H of acetaldehydes substituted with -CH3, -OH, -CH=CH2, and -C6H5 groups, respectively, are potential precursors of toxic nitriles and (N-chloro)amides during chloramination. Thus, more attention should be given to these aldehydes. The findings of this work are helpful for further understanding the aldehyde pathway mechanisms and predicting potential precursors of toxic nitriles and (N-chloro)amides during chloramination.

Identification of chlorinated products from tyrosine and tyrosyl dipeptides during chlorination: a computational study.

Chlorinated amino acids and peptides, as the model modified protein structures relevant to pathogen inactivation and an emerging class of disinfection byproducts (DBPs) with potential health risks to humans, have attracted much attention. However, due to a large variety of peptides (over 600) identified in source water and most of them featuring multiple reaction sites, it is a huge challenge to identify all the chlorinated amino acids and peptides. As a good complement to the experiment, quantum chemical computation can be used to uncover the chlorination sites and chlorinated products. In this study, frequently detected tyrosine (Tyr) and tyrosine-amide (Tyr-Am) as well as N-acetyl-tyrosine (NacTyr) were chosen as the model amino acid and model dipeptides, respectively. The results indicate that the kinetic reactivity order of reactive sites with estimated apparent rate constants (kobs-est, in M-1 s-1) is amino N (107-8) ≫ mono-chlorinated amino N (101-3) >/≈ phenol ortho-C (100-3) ≫ meta-C (10-3), and phenol ortho-C5 (102-3) > ortho-C3 (100-2) for dipeptides, while in thermodynamics, phenol C sites are more favorable than amino N sites. Moreover, due to the smaller differences of kobs-est values between the mono-chlorinated amino N and the phenol ortho-C sites in tyrosyl dipeptides compared to free Tyr, more kinds of C-chloro-tyrosyl dipeptides are likely to be generated. Additionally, a structure-kinetic reactivity relationship study reveals good correlations between lg kobs-est and NPA charges and BDEs of protons released from amino/hydroxyl groups in tyrosyl compounds rather than FED2 (HOMO). The results are helpful to further understand the reactivity of various reaction sites in peptides and identify chlorinated products from tyrosyl peptides during chlorination.

Carbon dioxide in the nitrosation of amine: catalyst or inhibitor?

Nitrosamines are a class of carcinogenic, mutagenic, and teratogenic compounds generally produced from the nitrosation of amine. This paper investigates the mechanism for the formation of nitrosodimethylamine (NDMA) from the nitrosation of dimethylamine (DMA) by four common nitrosating agents (NO(2)(-), ONOO(-), N(2)O(3), and ONCl) in the absence and presence of CO(2) using the DFT method. New insights are provided into the mechanism, emphasizing that the interactions of CO(2) with amine and nitrosating agents are both potentially important in influencing the role of CO(2) (catalyst or inhibitor). The role of CO(2) as catalyst or inhibitor mainly depends on the nitrosating agents involved. That is, CO(2) shows the catalytic effect when the weak nitrosating agent NO(2)(-) or ONOO(-) is involved, whereas it is an inhibitor in the nitrosation induced by the strong nitrosating agent N(2)O(3) or ONCl. To conclude, CO(2) serves as a "double-edged sword" in the nitrosation of amine. The findings will be helpful to expand our understanding of the pathophysiological and environmental significance of CO(2) and to develop efficient methods to prevent the formation of carcinogenic nitrosamines.

Degradation mechanisms of simple aliphatic amines under ozonation: a DFT study.

Aliphatic amines as common constituents of dissolved organic nitrogen (DON) exhibit high reactivity during ozonation; however, our understanding of their degradation mechanisms is very limited. In this study, methylamine (MA) and ethylamine (EA), as well as their secondary and tertiary amines (DMA, DEA, TMA and TEA) were chosen as aliphatic amine models and their degradation mechanisms during ozonation were investigated by using the DFT method. The oxygen-transfer reaction occurs initially and rapidly in the ozonation of all the above amines with a ΔG≠ value of 8-10 kcal mol-1 in great agreement with the experimental rate constant of 104 to 107 M-1 s-1. Moreover, N-oxide as the main degradation product for tertiary amines directly forms after oxygen-transfer, while nitroalkanes as main products for secondary and primary amines are yielded after a series of reactions mediated by hydroxylamine and nitrosoalkane with a ΔG≠ value of 10-13 kcal mol-1. Regarding the minor N-dealkylated products for all amines, alkylamino alcohol is an important intermediate possibly generated via a radical reaction pathway with a ΔG≠ value of 21-34 kcal mol-1. Additionally, comparison of the reactivity of aliphatic amines, hydroxylamines and alkylamino alcohols with ozone was made and elucidated in this study. The results are expected to expand our understanding of the degradation mechanisms for nitrogenous compounds during ozonation.