alkylating agents

Summary

Summary: Highly reactive chemicals that introduce alkyl radicals into biologically active molecules and thereby prevent their proper functioning. Many are used as antineoplastic agents, but most are very toxic, with carcinogenic, mutagenic, teratogenic, and immunosuppressant actions. They have also been used as components in poison gases.

Top Publications

  1. Gondo Y, Fukumura R, Murata T, Makino S. ENU-based gene-driven mutagenesis in the mouse: a next-generation gene-targeting system. Exp Anim. 2010;59:537-48 pubmed
    ..In the near future, next-generation re-sequencing systems should drastically enhance the utility of the ENU-based gene-driven mutagenesis not only for the mouse but also for other species. ..
  2. Zhou J, Ng S, Huang Q, Wu Y, Li Z, Yao S, et al. AMPK mediates a pro-survival autophagy downstream of PARP-1 activation in response to DNA alkylating agents. FEBS Lett. 2013;587:170-7 pubmed publisher
    ..Taken together, data from this study reveal a novel mechanism in controlling MNNG-mediated autophagy via AMPK activation downstream of PARP-1 activation and ATP depletion. ..
  3. Hata S, Hirayama J, Kajiho H, Nakagawa K, Hata Y, Katada T, et al. A novel acetylation cycle of transcription co-activator Yes-associated protein that is downstream of Hippo pathway is triggered in response to SN2 alkylating agents. J Biol Chem. 2012;287:22089-98 pubmed publisher
    ..the discovery of a novel cycle of acetylation/deacetylation of nuclear YAP induced in response to S(N)2 alkylating agents. We show that after treatment of cells with the S(N)2 alkylating agent methyl methanesulfonate, YAP ..
  4. Marey H, Mandour S, Ellakwa A. Subscleral trabeculectomy with mitomycin-C versus ologen for treatment of glaucoma. J Ocul Pharmacol Ther. 2013;29:330-4 pubmed publisher
    ..We conclude that the use of the Ologen implant in SST is comparable to the use of MMC with advantage of avoiding the potential dangerous complications related to MMC use in the early (12 months) follow-up period. ..
  5. Rippa V, Duilio A, di Pasquale P, Amoresano A, Landini P, Volkert M. Preferential DNA damage prevention by the E. coli AidB gene: A new mechanism for the protection of specific genes. DNA Repair (Amst). 2011;10:934-41 pubmed publisher
    aidB is one of the four genes of E. coli that is induced by alkylating agents and regulated by Ada protein. Three genes (ada, alkA, and alkB) encode DNA repair proteins that remove or repair alkylated bases...
  6. Löser D, Shibata A, Shibata A, Woodbine L, Jeggo P, Chalmers A. Sensitization to radiation and alkylating agents by inhibitors of poly(ADP-ribose) polymerase is enhanced in cells deficient in DNA double-strand break repair. Mol Cancer Ther. 2010;9:1775-87 pubmed publisher
    ..PARP inhibitors also enhance the cytotoxicity of ionizing radiation and alkylating agents but will only improve clinical outcomes if tumor sensitization exceeds effects on normal tissues...
  7. Chiu L, Ho F, Shiah S, Chang Y, Lin W. Oxidative stress initiates DNA damager MNNG-induced poly(ADP-ribose)polymerase-1-dependent parthanatos cell death. Biochem Pharmacol. 2011;81:459-70 pubmed publisher
    ..Moreover, intracellular calcium elevation and ROS production have mutual amplification effects and thus contribute to PARP-1-mediated parthanatos. ..
  8. Ström C, Johansson F, Uhlen M, Szigyarto C, Erixon K, Helleday T. Poly (ADP-ribose) polymerase (PARP) is not involved in base excision repair but PARP inhibition traps a single-strand intermediate. Nucleic Acids Res. 2011;39:3166-75 pubmed publisher
    ..Unexpectedly, addition of PARP inhibitor 2?h after DMS treatment still increased SSB levels indicating ongoing repair even at this late time point. ..
  9. Tsubura A, Lai Y, Miki H, Sasaki T, Uehara N, Yuri T, et al. Review: Animal models of N-Methyl-N-nitrosourea-induced mammary cancer and retinal degeneration with special emphasis on therapeutic trials. In Vivo. 2011;25:11-22 pubmed
    ..In this review, the targets of MNU in various animal species are described, and special emphasis is given to therapeutic trials against MNU-induced mammary cancer and retinal degeneration in animal models. ..

More Information

Publications62

  1. Furda A, Marrangoni A, Lokshin A, Van Houten B. Oxidants and not alkylating agents induce rapid mtDNA loss and mitochondrial dysfunction. DNA Repair (Amst). 2012;11:684-92 pubmed publisher
    ..These results suggest that persistent mtDNA damage is not sufficient to cause mitochondrial dysfunction. ..
  2. Fu D, Calvo J, Samson L. Balancing repair and tolerance of DNA damage caused by alkylating agents. Nat Rev Cancer. 2012;12:104-20 pubmed publisher
    b>Alkylating agents constitute a major class of frontline chemotherapeutic drugs that inflict cytotoxic DNA damage as their main mode of action, in addition to collateral mutagenic damage...
  3. Conde F, Ontoso D, Acosta I, Gallego Sánchez A, Bueno A, San Segundo P. Regulation of tolerance to DNA alkylating damage by Dot1 and Rad53 in Saccharomyces cerevisiae. DNA Repair (Amst). 2010;9:1038-49 pubmed publisher
    ..We propose that threshold levels of Rad53 activity exquisitely modulate the tolerance to alkylating damage at least by controlling the abundance of the key TLS factor Rev1 bound to chromatin. ..
  4. Knoche E, McLeod H, Graubert T. Pharmacogenetics of alkylator-associated acute myeloid leukemia. Pharmacogenomics. 2006;7:719-29 pubmed
    ..Identification of these susceptibility alleles should help clinicians develop tailored therapies that reduce the relative risk of t-AML. ..
  5. Cheng J, Xi G, Wei R, Cai J, Li Y. Efficacy and tolerability of nonpenetrating glaucoma surgery augmented with mitomycin C in treatment of open-angle glaucoma: a meta-analysis. Can J Ophthalmol. 2009;44:76-82 pubmed publisher
    ..However, significantly fewer patients achieved the target IOP with NPGS augmented with MMC than with TE plus MMC. NPGS augmented with MMC was better tolerated than TE plus MMC. ..
  6. Russell L, Hunsicker P, Russell W. Comparison of the genetic effects of equimolar doses of ENU and MNU: while the chemicals differ dramatically in their mutagenicity in stem-cell spermatogonia, both elicit very high mutation rates in differentiating spermatogonia. Mutat Res. 2007;616:181-95 pubmed
    ..g., chromosome aberrations, toxicity to germ cells and to animals, teratogenicity) revealed that while MNU is generally more effective, the opposite is true when the target cells are SG. ..
  7. Conde F, San Segundo P. Role of Dot1 in the response to alkylating DNA damage in Saccharomyces cerevisiae: regulation of DNA damage tolerance by the error-prone polymerases Polzeta/Rev1. Genetics. 2008;179:1197-210 pubmed publisher
    ..We propose that Dot1 inhibits translesion synthesis (TLS) by Polzeta/Rev1 and that the MMS resistance observed in the dot1 mutant results from the enhanced TLS activity. ..
  8. Meira L, Moroski Erkul C, Green S, Calvo J, Bronson R, Shah D, et al. Aag-initiated base excision repair drives alkylation-induced retinal degeneration in mice. Proc Natl Acad Sci U S A. 2009;106:888-93 pubmed publisher
    ..In wild-type animals DNA alkylating agents induce photoreceptor apoptosis and severe retinal degeneration...
  9. Palanca Capistrano A, Hall J, Cantor L, Morgan L, Hoop J, WuDunn D. Long-term outcomes of intraoperative 5-fluorouracil versus intraoperative mitomycin C in primary trabeculectomy surgery. Ophthalmology. 2009;116:185-90 pubmed publisher
    ..Both antifibrosis agents may contribute to the development of bleb leakage. The author(s) have no proprietary or commercial interest in any materials discussed in this article. ..
  10. Eker A, Quayle C, Chaves I, van der Horst G. DNA repair in mammalian cells: Direct DNA damage reversal: elegant solutions for nasty problems. Cell Mol Life Sci. 2009;66:968-80 pubmed publisher
    ..Part of a Multi-author Review). ..
  11. Liu X, Luo X, Shi Y, Zhu G, Penning T, Giranda V, et al. Poly (ADP-ribose) polymerase activity regulates apoptosis in HeLa cells after alkylating DNA damage. Cancer Biol Ther. 2008;7:934-41 pubmed
    ..Unlike in HeLa cells, ABT-888 protected MNNG induced cell death in normal human fibroblasts. Our study provides evidence that PARP activity determines the fate of HeLa cells by regulating the level of ATP after treatment with MNNG. ..
  12. Shigeeda T, Tomidokoro A, Chen Y, Shirato S, Araie M. Long-term follow-up of initial trabeculectomy with mitomycin C for primary open-angle glaucoma in Japanese patients. J Glaucoma. 2006;15:195-9 pubmed
    ..9+/-2.4%. Long-term outcome after initial trabeculectomy with MMC in Japanese POAG patients is comparable with that reported in other populations and with that after trabeculectomy with 5-fluorouracil. ..
  13. Sedgwick B, Bates P, Paik J, Jacobs S, Lindahl T. Repair of alkylated DNA: recent advances. DNA Repair (Amst). 2007;6:429-42 pubmed
    Cytotoxic and mutagenic methylated bases in DNA can be generated by endogenous and environmental alkylating agents. Such damaged bases are removed by three distinct strategies...
  14. Yang L, Li D, Chen J, Yang J, Xue L, Hu S, et al. Microarray expression analysis of the early N-methy-N-nitrosourea-induced retinal degeneration in rat. Neurosci Lett. 2007;418:38-43 pubmed
    ..These microarray results provide clues to understand the molecular pathways underlying photoreceptor degeneration and indicate directions for future studies. ..
  15. Staszewski O, Nikolova T, Kaina B. Kinetics of gamma-H2AX focus formation upon treatment of cells with UV light and alkylating agents. Environ Mol Mutagen. 2008;49:734-40 pubmed publisher
    ..Here we show that DNA damage induced by alkylating agents [methyl methanesulfonate (MMS) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)] and ultraviolet light (UV-C) ..
  16. Sakurai T, Maeda S, Chang L, Karin M. Loss of hepatic NF-kappa B activity enhances chemical hepatocarcinogenesis through sustained c-Jun N-terminal kinase 1 activation. Proc Natl Acad Sci U S A. 2006;103:10544-51 pubmed
  17. Kisby G, Olivas A, Park T, Churchwell M, Doerge D, Samson L, et al. DNA repair modulates the vulnerability of the developing brain to alkylating agents. DNA Repair (Amst). 2009;8:400-12 pubmed publisher
    ..the methylating agents methylazoxymethanol (MAM) and dimethyl sulfate (DMS) and the mono- and bifunctional alkylating agents chloroethylamine (CEA) and nitrogen mustard (HN2), in granule cell neurons derived from the cerebellum of ..
  18. van Boxtel R, Toonen P, van Roekel H, Verheul M, Smits B, Korving J, et al. Lack of DNA mismatch repair protein MSH6 in the rat results in hereditary non-polyposis colorectal cancer-like tumorigenesis. Carcinogenesis. 2008;29:1290-7 pubmed publisher
    ..The MSH6 knockout rat complements existing models for studying genetic instable tumorigenesis as it provides experimental opportunities that are not available or suboptimal in current models. ..
  19. Yanow S, Purcell L, Pradel G, Sato A, Rodriguez A, Lee M, et al. Potent antimalarial and transmission-blocking activities of centanamycin, a novel DNA-binding agent. J Infect Dis. 2008;197:527-34 pubmed publisher
    ..This demonstrates that damage to parasite DNA during blood-stage infection persists from the vertebrate to the mosquito host and provides a novel biochemical strategy to block malaria transmission. ..
  20. Moubarak R, Yuste V, Artus C, Bouharrour A, Greer P, Menissier de Murcia J, et al. Sequential activation of poly(ADP-ribose) polymerase 1, calpains, and Bax is essential in apoptosis-inducing factor-mediated programmed necrosis. Mol Cell Biol. 2007;27:4844-62 pubmed
    ..Our data shed new light on the mechanisms regulating AIF-dependent necrosis and support the notion that, like apoptosis, necrosis could be a highly regulated cell death program. ..
  21. Fenske T, McMahon C, Edwin D, Jarvis J, Cheverud J, Minn M, et al. Identification of candidate alkylator-induced cancer susceptibility genes by whole genome scanning in mice. Cancer Res. 2006;66:5029-38 pubmed
  22. Fry R, Svensson J, Valiathan C, Wang E, Hogan B, Bhattacharya S, et al. Genomic predictors of interindividual differences in response to DNA damaging agents. Genes Dev. 2008;22:2621-6 pubmed publisher
    ..Many proteins encoded by these genes are interconnected in cellular networks related to human cancer and tumorigenesis. ..
  23. Meador J, Zhao M, Su Y, Narayan G, Geard C, Balajee A. Histone H2AX is a critical factor for cellular protection against DNA alkylating agents. Oncogene. 2008;27:5662-71 pubmed publisher
    ..Cellular sensitivity to two monofunctional alkylating agents (methyl methane sulfonate and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)) was dependent on H2AX dosage, and ..
  24. Cledes G, Felizardo R, Foucart J, Carpentier P. Validation of a chemical osteoarthritis model in rabbit temporomandibular joint: a compliment to biomechanical models. Int J Oral Maxillofac Surg. 2006;35:1026-33 pubmed
    ..This could be interpreted as a repairing attempt due to the specific potential of fibrocartilage proliferative cells...
  25. Margison G, Butt A, Pearson S, Wharton S, Watson A, Marriott A, et al. Alkyltransferase-like proteins. DNA Repair (Amst). 2007;6:1222-8 pubmed
    ..their mechanism of action and role in protecting the host organisms against the biological effects of O(6)-alkylating agents, and their evolution...
  26. Homma K, Osakada F, Hirami Y, Jin Z, Mandai M, Takahashi M. Detection of localized retinal malfunction in retinal degeneration model using a multielectrode array system. J Neurosci Res. 2009;87:2175-82 pubmed publisher
    ..This spatial information should be valuable in investigating local functional recovery in therapeutically treated areas, such as in gene transfer or cell transplantation. ..
  27. Li H, Marple T, Hasty P. Ku80-deleted cells are defective at base excision repair. Mutat Res. 2013;745-746:16-25 pubmed publisher
    ..Here we show that Ku80 deletion also decreased resistance to ROS and alkylating agents that typically cause base lesions and single-strand breaks (SSBs)...
  28. Gómez Bombarelli R, Palma B, Martins C, Kranendonk M, Rodrigues A, Calle E, et al. Alkylating potential of oxetanes. Chem Res Toxicol. 2010;23:1275-81 pubmed publisher
    Small, highly strained heterocycles are archetypical alkylating agents (oxiranes, beta-lactones, aziridinium, and thiirinium ions)...
  29. Ekhart C, Rodenhuis S, Smits P, Beijnen J, Huitema A. An overview of the relations between polymorphisms in drug metabolising enzymes and drug transporters and survival after cancer drug treatment. Cancer Treat Rev. 2009;35:18-31 pubmed publisher
    ..survival in patients with different malignancies who are treated with substrates for these GSTs (mostly alkylating agents and platinum compounds)...
  30. Gonzalez Huici V, Wang B, Gartner A. A Role for the Nonsense-Mediated mRNA Decay Pathway in Maintaining Genome Stability in Caenorhabditis elegans. Genetics. 2017;206:1853-1864 pubmed publisher
    ..Hyper-sensitivity was not observed upon treatment with alkylating agents or UV irradiation...
  31. Held D, Shurtleff A, Fields S, Green C, Fong J, Jones R, et al. Vaccination of rabbits with an alkylated toxoid rapidly elicits potent neutralizing antibodies against botulinum neurotoxin serotype B. Clin Vaccine Immunol. 2010;17:930-6 pubmed publisher
  32. Lee M, Zhang S, Lin S, Chea J, Wang X, Leroy C, et al. Regulation of human DNA polymerase delta in the cellular responses to DNA damage. Environ Mol Mutagen. 2012;53:683-98 pubmed publisher
    The p12 subunit of polymerase delta (Pol ?) is degraded in response to DNA damage induced by UV, alkylating agents, oxidative, and replication stresses. This leads to the conversion of the Pol ?4 holoenzyme to the heterotrimer, Pol ?3...
  33. Luke A, Chastain P, Pachkowski B, Afonin V, Takeda S, Kaufman D, et al. Accumulation of true single strand breaks and AP sites in base excision repair deficient cells. Mutat Res. 2010;694:65-71 pubmed publisher
    ..Furthermore, the Pol ?-null cells displayed greater AP site formation than the parental DT40 cells. OTX use represents a facile approach for assessing SSB formation, whose benefits can also be applied to other established SSB assays. ..
  34. Mahdy R. Adjunctive use of bevacizumab versus mitomycin C with Ahmed valve implantation in treatment of pediatric glaucoma. J Glaucoma. 2011;20:458-63 pubmed publisher
    ..05). The adjunctive use of bevacizumab or mitomycin C during Ahmed valve implantation significantly enhances the valve survival and the first drug seems to be much safer with no visually devastating complications. ..
  35. Mul J, Nadra K, Jagalur N, Nijman I, Toonen P, Medard J, et al. A hypomorphic mutation in Lpin1 induces progressively improving neuropathy and lipodystrophy in the rat. J Biol Chem. 2011;286:26781-93 pubmed publisher
    ..Although we are cautious in making a direct parallel between the presented rodent model and human disease, our data may provide new insight into the pathogenicity of recently identified human LPIN1 mutations. ..
  36. Nikolova T, Ensminger M, Lobrich M, Kaina B. Homologous recombination protects mammalian cells from replication-associated DNA double-strand breaks arising in response to methyl methanesulfonate. DNA Repair (Amst). 2010;9:1050-63 pubmed publisher
    ..Further, they show that HR is the major pathway of protection of cells against DSB formation, killing and genotoxicity following S(N)2-alkylating agents.
  37. Trmčić M, Chadbourne F, Brear P, Denny P, Cobb S, Hodgson D. Aqueous synthesis of N,S-dialkylthiophosphoramidates: design, optimisation and application to library construction and antileishmanial testing. Org Biomol Chem. 2013;11:2660-75 pubmed publisher
    ..The hydrolysis properties of N-thiophosphoramidate ions and their reactivities towards alkylating agents are also explored...
  38. Pei D, Yang X, Liu W, Guikema J, Schrader C, Strauss P. A novel regulatory circuit in base excision repair involving AP endonuclease 1, Creb1 and DNA polymerase beta. Nucleic Acids Res. 2011;39:3156-65 pubmed publisher
    ..These results describe a novel regulatory circuit involving Apex, Creb1 and Polb and provide a mechanism for lethality of Apex loss in higher eukaryotes. ..
  39. Jacociunas L, de Andrade H, Lehmann M, de Abreu B, Ferraz A, da Silva J, et al. Artichoke induces genetic toxicity and decreases ethyl methanesulfonate-related DNA damage in Chinese hamster ovary cells. J Med Food. 2012;15:873-8 pubmed publisher
    ..However, the lowest dose was more protective than higher concentrations. Flavonoids and phenolic compounds are, probably, the C. scolymus constituents responsible for its genotoxic and antigenotoxic effects. ..
  40. Izumchenko E, Saydi J, Brown K. Exonuclease 1 (Exo1) is required for activating response to S(N)1 DNA methylating agents. DNA Repair (Amst). 2012;11:951-64 pubmed publisher
    ..This study indicates that EXO1 is required to activate MMR-dependent DDR in response to S(N)1 methylating agents; however, this function of EXO1 is independent of its nucleolytic activity. ..
  41. Ghoneim E, Abd El Hameed M. Needling augmented with topical application of mitomycin C for management of bleb failure. J Glaucoma. 2011;20:528-32 pubmed publisher
    ..Topical application of mitomycin C with needle revision seems to be an extremely effective way to revive failed filtration surgery. The incidence of complications related to mitomycin C was minimal. ..
  42. Kobbe S, Trapp O, Knoll A, Manuss A, Puchta H. The Translesion Polymerase ζ Has Roles Dependent on and Independent of the Nuclease MUS81 and the Helicase RECQ4A in DNA Damage Repair in Arabidopsis. Plant Physiol. 2015;169:2718-29 pubmed publisher
    ..processivity is mainly required for resistance to interstrand and intrastrand cross-linking agents, but not alkylating agents. To better define the role of REV3 in relation to other key factors involved in DNA repair, we perform ..
  43. Fedhila Ben Ayed F, Rhayem S, Doghri R, Ben Hassine L, Khemiri M, Mrad K, et al. [Secondary TFE3-associated renal cell carcinoma in a child treated for Ewing sarcoma]. Arch Pediatr. 2016;23:171-5 pubmed publisher
    ..It seems that translocation renal cell carcinoma is associated with previous exposure to chemotherapy, particularly topoisomerase II inhibitors or alkylating agents.
  44. Lv D, Yu S, Ping Y, Wu H, Zhao X, Zhang H, et al. A three-dimensional collagen scaffold cell culture system for screening anti-glioma therapeutics. Oncotarget. 2016;7:56904-56914 pubmed publisher
    ..3D-cultured cells also exhibited enhanced resistance to chemotherapeutic alkylating agents, with a much higher proportion of glioma stem cells and upregulation of O6-methylguanine DNA ..
  45. Promkum C, Kupradinun P, Tuntipopipat S, Butryee C. Nutritive evaluation and effect of Moringa oleifera pod on clastogenic potential in the mouse. Asian Pac J Cancer Prev. 2010;11:627-32 pubmed
    ..In conclusion, the present study demonstrated that bMO has no clastogenicity and possesses anticlastogenic potential against clastogens, and particularly a direct-acting clastogen in the mouse. ..
  46. Mlambo G, Coppens I, Kumar N. Aberrant sporogonic development of Dmc1 (a meiotic recombinase) deficient Plasmodium berghei parasites. PLoS ONE. 2012;7:e52480 pubmed publisher
    ..Our findings suggest that PbDmc1 plays a critical role in malaria transmission biology. ..
  47. Cravedi P, Angeletti A, Remuzzi G. New biologics in the treatment of rare glomerular diseases of childhood. Curr Opin Pharmacol. 2017;33:27-33 pubmed publisher
    ..the use of immunosuppressive drugs, such as corticosteroids, calcineurin inhibitors, antiproliferative and alkylating agents. Unfortunately, these treatments fail to achieve a sustained remission in a significant portion of patients ..
  48. Mateos M, San Miguel J. Management of multiple myeloma in the newly diagnosed patient. Hematology Am Soc Hematol Educ Program. 2017;2017:498-507 pubmed publisher
  49. Pabla N, Ma Z, McIlhatton M, Fishel R, Dong Z. hMSH2 recruits ATR to DNA damage sites for activation during DNA damage-induced apoptosis. J Biol Chem. 2011;286:10411-8 pubmed publisher
    ..Together the results support a hMSH2-dependent pathway of ATR activation and downstream Chk2/p53 signaling. ..
  50. Shojaei A, Ramezanzadeh M, Soleyman Jahi S, Almasi Nasrabadi M, Rezazadeh P, Eslani M. Short-time mitomycin-C application during photorefractive keratectomy in patients with low myopia. J Cataract Refract Surg. 2013;39:197-203 pubmed publisher
    ..No author has a financial or proprietary interest in any material or method mentioned. ..
  51. Eberwein P, Böhringer D, Schwartzkopff J, Birnbaum F, Reinhard T. Allogenic limbo-keratoplasty with conjunctivoplasty, mitomycin C, and amniotic membrane for bilateral limbal stem cell deficiency. Ophthalmology. 2012;119:930-7 pubmed publisher
  52. Hu X, Yu Y, Li Q, Wu D, Tan Z, Wang C, et al. Quantitative and subcellular localization analysis of the nuclear isoform dUTP pyrophosphatase in alkylating agent-induced cell responses. Toxicology. 2011;287:15-20 pubmed publisher
    ..These results suggest that DUT-N may be a potential early biomarker to assess the risk of alkylating agents exposure. DUT-N is one of the two isoforms of deoxyuridine triphosphate nucleotidohydrolase (dUTPase)...
  53. Deeley R, Westlake C, Cole S. Transmembrane transport of endo- and xenobiotics by mammalian ATP-binding cassette multidrug resistance proteins. Physiol Rev. 2006;86:849-99 pubmed
    ..analogs, arsenical and antimonial oxyanions, peptide-based agents, and, under certain circumstances, alkylating agents. The MRPs are also primary active transporters of other structurally diverse compounds, including ..