Journal of Nanomaterials

Journal of Nanomaterials / 2015 / Article

Research Article | Open Access

Volume 2015 |Article ID 374218 | 8 pages |

In Silico Study of Spacer Arm Length Influence on Drug Vectorization by Fullerene C60

Academic Editor: Miguel A. Correa-Duarte
Received12 Jan 2015
Revised09 May 2015
Accepted10 May 2015
Published04 Jun 2015


This work studies theoretically the effect of spacer arm lengths on the characteristics of a fullerene C60-based nanovector. The spacer arm is constituted of a carbon chain including a variable number of methylene groups (n = 2–11). To improve the ability of the fullerene carriage, two arms are presented simultaneously through a malonyl bridge. Then the evolution of selected physicochemical parameters is monitored as a function of the spacer arm length and the angle between the two arms. We show here that while the studied characteristics are almost independent of the spacer arm length or vary monotonically with it, the dipole moment and its orientation vary periodically with the parity of the number of carbon atoms. This periodicity is related to both modules and orientations of dipole moments of the spacer arms. In the field of chemical synthesis, these results highlight the importance of theoretical calculations for the optimization of operating conditions. In the field of drug discovery, they show that theoretical calculations of the chemical properties of a drug candidate can help predict its in vivo behaviour, notably its bioavailability and biodistribution, which are known to be tightly dependent of its polarity.

1. Introduction

At the beginning of the 21st century, the fight against cancer remains one of the major public health problems. This is mainly due to some impediments that hinder the achievement of significant therapeutic progress, mainly toxic adverse effects, cell resistance to drugs [1], and high cost of research. A promising way to overcome these obstacles consists in using natural products or some of their extracts. On another hand, drug vectorization and targeting are now recognized as the best way for modulating the bioavailability and thus the toxic adverse effects of an anticancer agent.

Among the different platforms for vectoring active principles (AP) Fullerene C60 [24] is of particular interest because this carbon nanostructured compound has very interesting biomedical features. Indeed, the toxicity of C60 is now well understood [5] and its beneficial health effects encompass a large variety of biomedical fields including imaging, photodynamic therapy, gene delivery [6], oxidative stress [3, 4], and even life extension [79].

There are two possible vectorization routes: encapsulation of the AP within the vector or its grafting on the outer walls. The aim of this study is to explore theoretically the vectorization process by grafting thymoquinone (TQ), a model of anticancer natural product, on a fullerene molecule, which is characterized by its nanometric size, high lipophilicity, and chemical reactivity [8].

Thymoquinone (TQ), the most abundant component of black seeds (Nigella sativa), has been used for centuries in the Middle East as a natural medicine for the treatment of many diseases [10, 11]. Several pharmacological studies have demonstrated that TQ is able to develop antioxidant, anti-inflammatory, and antineoplastic effects in vitro as well as in vivo against various tumor cells [1215]. Nowadays, TQ has attracted considerable interest and many reports have shown that the inhibitory effects of this compound are specific of cancer cells, including those of breast, prostate, and pancreas cells [14, 16, 17]. A previous report also showed a higher anticancer activity for nanoparticle encapsulated TQ than for free TQ due to improved cellular uptake and bioavailability [18], as in vitro experiments have shown that the activity of a vectorized drug and the successful operation of vectorization strongly depend on the type of linker used [1921].

Polarity is an important characteristic of a molecule because it can influence its chemical, physical, and biological properties. Some in vitro studies [23] on the activities of a series of histamine H2 antagonists have shown that relatively minor structural changes greatly influence the antagonistic activity. Because the molecules were very polar, these differences were attributed to the orientations of the dipole moment and interpreted by the relative alignment of hydrogen bonding. For peptides in Aib homooligomers with one, two, and three intramolecular hydrogen bonds, dipole moments increase with the number of Aib units by roughly 2.3 D/residue. The obtained values are 8.22, 10.79, and 12.34 D, respectively [24].

In the research work of Garbuio et al., [25] two series of compounds formed by a nitroxide radical linked by a peptide bond to the fullerene C60 are similar to those of our calculated compounds, the authors have shown that the direction of the resulting molecular dipole moment could be changed by reversing the position of fullerene and nitroxide with respect to the nitrogen of the peptide. The electrochemical analysis and chemical nitroxide reduction experiments indicate that the dipole moment significantly affects the redox properties of the two electroactive groups.

Our calculations are designed to study the characteristics and properties of the PA TQ graft on the C60 through bra formed by a carbon chain of variable length –(CH2)n ( = 2–11). This is a relatively inert motif that minimizes interaction with different parts of the human body during drug transport. Its variable length could allow better control of the overall size and physicochemical properties of the studied nanocarriers, which are important parameters for their therapeutic activity.

2. Computational Details

Calculations are computed by the Gaussian [26, 27] packet and recovered by GAUSSVIEW program [28]. Hartree-Fock (HF) and density functional theory (DFT/B3LYP) methods are used for geometrical optimization. Given the great number of atoms forming the studied compound and in order to keep the calculations time compatible with our machines, basis sets STO-3G of moderate dimension were used. For small compounds, the basis set 6-311G(d,p) [29] has also been tested. We checked that all vibrational frequencies are positive, which indicates that each studied structure coincides with a minimum on the potential surface.

3. Results and Discussion

DFT technique has been shown to be reliable and commonly used for the functional study of various nanostructures [30, 31]. The studied compound has a four-part structure consisting of (i) TQ, the active principle, (ii) a spacer arm, (iii) a connecting bridge, and (iv) C60 molecule, the drug carrier. To validate our computational techniques and choice of basis sets, we first studied the free TQ molecule. In the second section of this work we focused on the spacer arm and the bridge. The final part deals with the drug candidate that can be synthesized and used in medicine.

3.1. Thymoquinone

Thymoquinone is the 2-isopropyl-5-methyl-1,4-benzoquinone whose molecular formula is C10H12O2 corresponding to two geometrical isomers [32, 33] (Figure 1).

The structure of TQ is optimized in order to enable the localization of any geometrical change induced by its binding to C60. In order to maintain the therapeutic potential of TQ, it is mandatory to avoid any structural modification after its binding to C60.

The calculated values of the geometrical parameters are shown in Table 1. These values are in accordance with those calculated by other authors [22]. Two stable conformations with C1 (trans) and Cs (cis) symmetries (Figure 1) were identified.


TQ transTQ + C60
Geometrical parametersSTO-3G6-31G(d)6-311++G(d,p)STO-3G
HFB3LYPB3LYP [22]MPW1PW91 [22]MP2 [22]B3LYP [22]MPW1PW91 [22]HF

Distances (Å)
Angles (°)


TQ Cis
Geometrical parametersSTO-3G6-31G(d)6-311++G(d,p)
HFB3LYPB3LYP [22]MPW1PW91 [22]MP2 [22]B3LYP [22]MPW1PW91 [22]

Distances (Å)
Angles (°)

Theoretical calculations clearly show that the transconformation is more stable than the cis one. Thus, we continued the calculations by using the former one. The transisomer is not a very polar compound because its dipole moment is only equal to 0.336 and 0.284 D as calculated with HF/STO-3G and DFT/6-311G(d,p), respectively.

3.2. The Spacer Arm

For the spacer arm, calculations were performed by both HF and DFT. The comparison between the two results was used for results validation in order to compensate the lack of experimental values for these compounds. Each spacer arm is formed by a saturated carbon chain and the number of methylene groups (–CH2) ranged from 2 to 11. At the chain ends, two functions containing OH groups (one primary alcohol and one carboxylic acid) were fixed to be easily substituted by chlorine atoms. The length of the arm is measured between these two fragments (Figure 2). Given the importance of the polarity in the interaction of TQ with the human body [34], a particular attention has been paid to the dipole moment.

Figure 3 shows the variation of the optimized energy as a function of the spacer arm length. By using HF and DFT methods, we obtained a linear relationship between both parameters ( ) and (), resp.), thus demonstrating that all studied spacer arms have similar behaviour.

In a multistage synthesis process, the yield depends on the correct choice of operating conditions particularly on the choice of the appropriate solvent. For these reasons, a special attention was given to the dipole moments of all studied compounds. Table 2 summarizes the dipole moment values data. These results show large variations as a function of the length of the spacer arm. A periodic phenomenon is observed, with maxima corresponding to even unit numbers of methylene (–CH2) and minima associated with odd numbers (Figure 4).

CompoundSpacer arm length (Å) Dipole moment (D)


The maxima are relatively high, which is consistent with the presence of polar moieties such as carboxylic acids and alcohols. For instance, Furylfulgide (Aberchrome 540) is known to exhibit an experimental dipole moment of 7.2 D [35].

As periodicity is observed, dipole moments values take substantially two different levels depending on the parity of the number of the spacer arm carbon atoms. The figures obtained from the vector shape of dipole moments also show alternating orientation of direction. Each dipole moment is drawn from the electronic barycenter.

To interpret both values and changes in dipole moments, we proceeded step by step. First the arm was divided into three parts, which are alcohol, carboxylic acid, and the remaining CH2 chain. To take reciprocal fragments interaction into account, we then computed each dipole moment separately with the remaining atoms being replaced by dummy ones as illustrated in Figure 5. So the three calculated dipole moments will have the same representation referential. Hence, we started with the shortest odd number of methylene group (). Given that one CH2 is included in the alcohol function, the remaining carbon motif –CH2–CH2– is apolar by symmetry. Adding dipole moments of fragments leads to 3.34 D, which is very close to the DFT calculated value of 3.95 D.

Details of calculations are summarized in Table 3. Roughly the same dipole moment value is obtained for all similar compounds with odd number of methylene groups. This is due to the fact that the central motif is not involved in the final dipole moment, as its partial moment is equal to zero. Moving now to the shortest arm that contains only a single methylene group, a carboxylic acid, and an alcoholic function. Here, the CH2 group alone becomes polar. Its dipole moment is 2.41 D. One of the other two moments will necessarily change in direction compared to the previous case with odd . The vector sum of three contributions changes in direction and value. So we get a value of 0.42 D, again close to the DFT calculated one of 0.41 D. A similar reasoning can be extended to all compounds with an even number of CH2 groups. To summarize these observations, whenever a methylene group is added to the chain, one of the two ended polar fragment moments changes in direction. Thus, the addition and the subtraction of their dipole moment will be alternated.

Carboxylic acidAlcoholCH2





DFT calculated value for the studied compound.

Given that the DFT technique provides consistent results with those obtained by the HF method results, the latter was chosen for the rest of the calculations as it requires much less computing time. When the number of studied atom patterns is large, theoretical calculations are performed to determine the optimized structures with the different spacer arms attached to C60 on three levels: small ( = 2), medium ( = 5), and large ( = 11). In order to increase the efficiency of the proposed protocol, two spacer arms are grafted on a support to connect them simultaneously to the fullerene molecule. The condensation of each spacer arm with malonyl dichloride allows reaching a stable adduct. Hydroxyl groups of the carboxylic end are substituted by chlorine atoms in order to prepare the final compound for the following step of condensation on TQ. Figure 6 depicts the descriptors and and the β1 angle.

The dipole moment of the diagram (Figure 7) is a function of the chlorinated compound length. There is a periodicity of dipole moment as a function of the spacer arm length. However, an inversion of the dipole moment is observed with respect to the arm alone. The highest values are obtained for odd numbers while the lowest values are linked to even ones. The HF results of the descriptors 2, 3, and 1 for the chlorinated arms are summarized in Table 4.

(Å) (Å) (D) (°)


3.3. The Final Compound [Arm-TQ]

Theoretical HF calculations were performed to determine the optimized structures of the various compounds of arm spacer’s condensation, along with different lengths, with TQ (Figure 8).

The relationship between the distance and the angle of the arm shown in Figure 9 obeys the following equation (Table 5): ().

(Å) (Å) (D) (°)


The angle of the two support arms decreases when the distance from the spacer arm increases.

3.4. Grafting the Final Compound [Arm-TQ] on C60

The effect of grafting on TQ structure and the experimental geometric parameters for the free TQ grafted onto fullerene C60 are summarized in Table 1(b). Small changes, in the 1% range, are observed. This demonstrates the conservation of the original structure of TQ thus meeting the first requirement of the vectorization process. Figure 10 shows the final compound corresponding to TQ grafted onto C60 via a spacer arm and its descriptors and (Table 6).

Compounds (Å) (Å) (D)


4. Conclusion

In this paper, we studied the effects of the spacer arm length on the synthesis conditions of a fullerene C60-based drug-vector. As a drug sample, we selected thymoquinone, a natural product with anticancer properties. The spacer arm was chosen for its biocompatibility since it is composed of a carbon chain including a variable number of methylene groups ( = 2–11) ending with alcohol and carboxylic acid functions. To improve the ability of the fullerene carriage, two arms were grafted simultaneously through a malonyl bridge [36].

All parts of the resulting nanosystem were studied separately. Their geometry was optimized and selected physicochemical parameters were calculated. The evolution of these parameters was monitored as a function of the spacer arm length and the angle between the two arms. While all the studied characteristics were almost independent of the spacer arm length or varied monotonically with it, the dipole moment exhibited periodicity depending on the parity of the number of carbon atoms in the chain. All other studied compounds exhibited the same periodic behaviour. This phenomenon is explained by the alternation of vector addition/subtraction when the parity of carbon atoms number was changed.

In the field of chemical synthesis, these results highlight the importance of theoretical calculations for the optimization of operating conditions. Indeed, the knowledge of chemical properties, notably the polarity of synthesised products and intermediates, is mandatory for the right choice of the solvents. In the field of C60-derivatives synthesis, the rule “like dissolves like” remains of high relevance. Indeed, C60’s solubility is known to be very sensitive to the polarity of the solvent. For instance, its solubility in 1-chloro-naphthalene ( = 50 mg·mL−1) is considerably higher than in methanol ( = 0.01 mg·mL−1) [37]. Hence, changes in the parity of the spacer arm will have a great significance in the synthesis of a C60 based nanovector.

As the synthesis process requires several steps, we have to find the appropriate solvent for each combination step that means that the solvent polarity should be controlled according to the parity of the spacer arm and the polarity of each synthesis product or synthesis intermediate. Finally, our results show that theoretical calculations of the chemical properties of a drug candidate can help predict its in vivo behaviour, notably its bioavailability and biodistribution, which are known to be tightly dependent of its polarity.

Conflict of Interests

The authors declare that there is no conflict of interests regarding to the publication of this paper.


  1. J. Davies, “Inactivation of antibiotics and the dissemination of resistance genes,” Science, vol. 264, no. 5157, pp. 375–382, 1994. View at: Publisher Site | Google Scholar
  2. H. W. Kroto, J. R. Heath, S. C. O'Brien, R. F. Curl, and R. E. Smalley, “C60: buckminsterfullerene,” Nature, vol. 318, no. 6042, pp. 162–163, 1985. View at: Publisher Site | Google Scholar
  3. A. W. Jensen, S. R. Wilson, and D. I. Schuster, “Biological applications of fullerenes,” Bioorganic and Medicinal Chemistry, vol. 4, no. 6, pp. 767–779, 1996. View at: Publisher Site | Google Scholar
  4. H. Szwarc and F. Moussa, “[60] Fullerene and derivatives in biology and medicine,” in Handbook of Fullerene: Synthesis, Properties and Applications, R. F. Verner and C. Benvegn, Eds., pp. 403–420, Nova Science Publishers, New York, NY, USA, 2012. View at: Google Scholar
  5. J. Kolosnjaj, H. Szwarc, and F. Moussa, “Toxicity studies of fullerenes and derivatives,” Advances in Experimental Medicine and Biology, vol. 620, pp. 168–180, 2007. View at: Publisher Site | Google Scholar
  6. R. Maeda-Mamiya, E. Noiri, H. Isobe et al., “In vivo gene delivery by cationic tetraamino fullerene,” Proceedings of the National Academy of Sciences of the United States of America, vol. 107, no. 12, pp. 5339–5344, 2010. View at: Publisher Site | Google Scholar
  7. K. L. Quick, S. S. Ali, R. Arch, C. Xiong, D. Wozniak, and L. L. Dugan, “A carboxyfullerene SOD mimetic improves cognition and extends the lifespan of mice,” Neurobiology of Aging, vol. 29, no. 1, pp. 117–128, 2008. View at: Publisher Site | Google Scholar
  8. J. Gao, Y. Wang, K. M. Folta et al., “Polyhydroxy fullerenes (fullerols or fullerenols): beneficial effects on growth and lifespan in diverse biological models,” PLoS ONE, vol. 6, no. 5, Article ID e19976, 2011. View at: Publisher Site | Google Scholar
  9. T. Baati, F. Bourasset, N. Gharbi et al., “The prolongation of the lifespan of rats by repeated oral administration of [60]fullerene,” Biomaterials, vol. 33, no. 19, pp. 4936–4946, 2012. View at: Publisher Site | Google Scholar
  10. M. S. Butt and M. T. Sultan, “Nigella sativa: reduces the risk of various maladies,” Critical Reviews in Food Science and Nutrition, vol. 50, no. 7, pp. 654–665, 2010. View at: Publisher Site | Google Scholar
  11. N. Ilaiyaraja and F. Khanum, “Nigella sativa L.: a review of therapeutic application,” Journal of Herbal Medicine and Toxicology, vol. 4, pp. 1–8, 2010. View at: Google Scholar
  12. S. Padhye, S. Banerjee, A. Ahmad, R. Mohammad, and F. H. Sarkar, “From here to eternity—the secret of Pharaohs: therapeutic potential of black cumin seeds and beyond,” Cancer Therapy, vol. 6, no. b, pp. 495–510, 2008. View at: Google Scholar
  13. C. C. Woo, A. P. Kumar, G. Sethi, and K. H. B. Tan, “Thymoquinone: potential cure for inflammatory disorders and cancer,” Biochemical Pharmacology, vol. 83, no. 4, pp. 443–451, 2012. View at: Publisher Site | Google Scholar
  14. M. N. Nagi and M. A. Mansour, “Protective effect of thymoquinone against doxorubicin-induced cardiotoxicity in rats: a possible mechanism of protection,” Pharmacological Research, vol. 41, no. 3, pp. 283–289, 2000. View at: Publisher Site | Google Scholar
  15. O. A. Al-Shabanah, O. A. Badary, M. N. Nagi, N. M. Al-Gharably, A. C. Al-Rikabi, and A. M. Al-Bekairi, “Thymoquinone protects against doxorubicin-induced cardiotoxicity without compromising its antitumor activity,” Journal of Experimental and Clinical Cancer Research, vol. 17, no. 2, pp. 193–198, 1998. View at: Google Scholar
  16. A. E. Edris, “Anti-cancer properties of Nigella spp. essential oils and their major constituents, thymoquinone and β-elemene,” Current Clinical Pharmacology, vol. 4, no. 1, pp. 43–46, 2009. View at: Publisher Site | Google Scholar
  17. R. Keyhanmanesh, M. H. Boskabady, S. Khamneh, and Y. Doostar, “Effect of thymoquinone on the lung pathology and cytokine levels of ovalbumin-sensitized guinea pigs,” Pharmacological Reports, vol. 62, no. 5, pp. 910–916, 2010. View at: Publisher Site | Google Scholar
  18. J. Ravindran, H. B. Nair, B. Sung, S. Prasad, R. R. Tekmal, and B. B. Aggarwal, “Thymoquinone poly (lactide-co-glycolide) nanoparticles exhibit enhanced anti-proliferative, anti-inflammatory, and chemosensitization potential,” Biochemical Pharmacology, vol. 79, no. 11, pp. 1640–1647, 2010. View at: Publisher Site | Google Scholar
  19. S. Kiyonaka, K. Sada, I. Yoshimura, S. Shinkai, N. Kato, and I. Hamachi, “Semi-wet peptide/protein array using supramolecular hydrogel,” Nature Materials, vol. 3, no. 1, pp. 58–64, 2004. View at: Publisher Site | Google Scholar
  20. S. J. Lee and S. Y. Lee, “Microarrays of peptides elevated on the protein layer for efficient protein kinase assay,” Analytical Biochemistry, vol. 330, no. 2, pp. 311–316, 2004. View at: Publisher Site | Google Scholar
  21. G. J. Wegner, H. J. Lee, and R. M. Corn, “Characterization and optimization of peptide arrays for the study of epitope-antibody interactions using surface plasmon resonance imaging,” Analytical Chemistry, vol. 74, no. 20, pp. 5161–5168, 2002. View at: Publisher Site | Google Scholar
  22. A. B. Raschi, E. Romano, A. M. Benavente, A. B. Altabef, and M. E. Tuttolomondo, “Structural and vibrational analysis of thymoquinone,” Spectrochimica Acta—Part A: Molecular and Biomolecular Spectroscopy, vol. 77, no. 2, pp. 497–505, 2010. View at: Publisher Site | Google Scholar
  23. R. C. Young, G. J. Durant, J. C. Emmett et al., “Dipole moment in relation to H2 receptor histamine antagonist activity for cimetidine analogues,” The Journal of Medicinal Chemistry, vol. 29, no. 1, pp. 44–49, 1986. View at: Google Scholar
  24. A. H. Holm, M. Ceccato, R. L. Donkers, L. Fabris, G. Pace, and F. Maran, “Effect of peptide ligand dipole moments on the redox potentials of Au 38 and Au140 nanoparticles,” Langmuir, vol. 22, no. 25, pp. 10584–10589, 2006. View at: Publisher Site | Google Scholar
  25. L. Garbuio, S. Antonello, I. Guryanov et al., “Effect of orientation of the peptide-bridge dipole moment on the properties of fullerene-peptide-radical systems,” Journal of the American Chemical Society, vol. 134, no. 25, pp. 10628–10637, 2012. View at: Publisher Site | Google Scholar
  26. M. J. Frisch, G. W. Trucks, H. B. Schlegel et al., Gaussian 03, Revision C. 02, Gaussian, Wallingford, Conn, USA, 2004.
  27. M. J. Frisch, G. W. Trucks, H. B. Schlegel et al., Gaussian 09 Revision A.1, Gaussian, Wallingford, Conn, USA, 2009.
  28. A. Frisch, A. B. Nielsen, and A. J. Holder, GAUSSIANVIEW Users Manual, Gaussian, Pittsburgh, Pa, USA, 2000.
  29. R. Ditchfield, W. J. Hehre, and J. A. Pople, “Self-consistent molecular orbital methods. 9. Extended Gaussian-type basis for molecular-orbital studies of organic molecules,” The Journal of Chemical Physics, vol. 54, no. 2, pp. 724–728, 1971. View at: Google Scholar
  30. A. Ahmadi, N. L. Hadipour, M. Kamfiroozi, and Z. Bagheri, “Theoretical study of aluminum nitride nanotubes for chemical sensing of formaldehyde,” Sensors and Actuators B: Chemical, vol. 161, no. 1, pp. 1025–1029, 2012. View at: Publisher Site | Google Scholar
  31. A. Ahmadi, J. Beheshtian, and N. L. Hadipour, “Chemisorption of NH3 at the open ends of boron nitride nanotubes: a DFT study,” Structural Chemistry, vol. 22, no. 1, pp. 183–188, 2011. View at: Publisher Site | Google Scholar
  32. P. J. Houghton, R. Zarka, B. de las Heras, and J. R. S. Hoult, “Fixed oil of Nigella sativa and derived thymoquinone inhibit eicosanoid generation in leukocytes and membrane lipid peroxidation,” Planta Medica, vol. 61, no. 1, pp. 33–36, 1995. View at: Publisher Site | Google Scholar
  33. N. Chakravarty, “Inhibition of histamine release from mast cells by nigellone,” Annals of Allergy, vol. 70, no. 3, pp. 237–242, 1993. View at: Google Scholar
  34. O. A. Badary, O. A. Al-Shabanah, M. N. Nagi, A. C. Al-Rikabi, and M. M. A. Elmazar, “Inhibition of benzo(a)pyrene-induced forestomach carcinogenesis in mice by thymoquinone,” European Journal of Cancer Prevention, vol. 8, no. 5, pp. 435–440, 1999. View at: Publisher Site | Google Scholar
  35. J. A. Delaire and K. Nakatani, “Linear and nonlinear optical properties of photochromic molecules and materials,” Chemical Reviews, vol. 100, no. 5, pp. 1817–1845, 2000. View at: Publisher Site | Google Scholar
  36. C. Bingel, “Cyclopropanierung von fullerenen,” Chemische Berichte, vol. 126, no. 8, pp. 1957–1959, 1993. View at: Publisher Site | Google Scholar
  37. R. S. Ruoff, D. S. Tse, R. Malhotra, and D. C. Lorents, “Solubility of C60 in a variety of solvents,” The Journal of Physical Chemistry, vol. 97, no. 13, pp. 3379–3383, 1993. View at: Publisher Site | Google Scholar

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