Development of a Sun Protection Factor for contact lenses (CL-SPF)

Abstract

Objective

Sun Protection Factor (SPF) of sunscreen products is well recognised by consumers. This study explored how SPF could be applied to ultraviolet radiation (UV) protection from contact lenses (CL-SPF).

Methods and analysis

UV transmission through 15 commercially available contact lenses and three spectacle lens materials was measured with a deuterium light source and spectrophotometer. CL-SPF values were calculated using the standard in vitro method used to test and label skin products. Ray tracing was applied to two sunglass designs to assess the effect of solar angle and head orientation on light reaching the ocular surface. Cellular damage profile of human corneal and conjunctival cells across the UV range was assessed in vitro to inform an SPF equivalent for CLs.

Results

CLs tested fell into three categories: CL-SPF with no UV blocker=1.0–2.0 (equivalent to using no sunscreen); CL-SPF with Class 2 UV blocker=12.3–24.8 (equivalent to SPF15); and CL-SPF with Class 1 UV blocker=59.6–66.2 (equivalent to SPF 50+). Despite the UV-blocking characteristics of sunglasses, ocular surface protection can be substantially reduced at certain solar angle and head orientation combinations; on average, 76%–89% of light was prevented from reaching the ocular surface depending on the intensity of the tint (80%–20% transmission). The data also suggest that cell damage and death of ocular surface cells has a similar profile to that of the skin, but conjunctival cells are more susceptible to UV damage.

Conclusion

CL-SPF is a viable metric to communicate the protection from the absorption/transmission of UV radiation that CLs offer wearers. However, a contact lens will only protect the area of the ocular surface it covers, which is limited to mainly the cornea and internal eye tissues with soft CLs.

WHAT IS ALREADY KNOWN ON THIS TOPIC

• While Sun Protection Factor (SPF) is used to rate the protection from ultraviolet radiation (UV) provided by sunscreen and some clothes, there is no equivalent communication tool for optical eye corrections.

WHAT THIS STUDY ADDS

• Confirmation is provided that the damage profile across UV wavelengths is similar to skin and this is applied to the UV transition of contact lenses and spectacle materials to calculate a contact lens-SPF for these products.

HOW THIS STUDY MIGHT AFFECT RESEARCH, PRACTICE OR POLICY

• This approach allows the contact lens industry to provide a more meaningful metric of the sun protection of their products, to aid the consumer.

Introduction

A primary role of eye care practitioners is to make their patients aware of avoidable risks to ocular health. While the importance of protecting the skin against the adverse effects of sunlight exposure is well recognised, the ocular effects of solar radiation and the need for eye protection are less well understood by patients.¹ Responses of ocular tissues to ultraviolet radiation (UV), the unique aspects of ocular exposure and the various methods of eye protection have been a focus for research.² Contact lenses (CLs) which incorporate UV-blocking help prevent UV transmission.^3–6 However, simple and effective methods of communicating the protective effect of UV-blocking CLs, and the differences between lenses with respect to UV protection, are lacking.

CLs that offer protection from the transmission of UV radiation can be labelled as Class 1 or Class 2, indicating the level of blocking. As specified by the International Standards Organisation (ISO), Class 1 must block at least 90% of UVA and at least 99% UVB and Class 2 must block at least 50% UVA and at least 95% UVB.⁷ This is based on the mean spectral transmission. However, consumers wearing lenses may find such terminology confusing and could interpret these per cent as the amount, that is, transmitted through the lens rather than blocked.

Sunglasses have a filter category from 0 to 4, where 0 has a luminous transmittance >80% and 4 has from 3% to 8% luminous transmittance as well as lateral protection to protect against the peripheral light focusing effect.⁸ However, when wearing sunglasses, light can still reach the internal eye through various routes, such as around the spectacle lens, by reflection off the back surface of the lens and through a dilated pupil, and this may also be magnified by the peripheral light focusing effect.⁹ Protection afforded by sunglasses and spectacle lenses is also highly dependent on solar angle and light orientation.¹⁰

One manufacturer (Essilor) introduced an ‘eye-sun protection factor’ (E-SPF) for clear spectacle lenses and sunglasses similar to the SPF factor on sunscreen products which is said to take account of UV transmittance and reflectance, corneal sensitivity to UV and solar spectral irradiance, solar angle effects and frame and lens parameters.^{11 12} As with skin products, the E-SPF is a ratio between eye exposure with and without protection.

The SPF of sunscreen products is commonly used to communicate protection from UV with consumers. Sunscreens are categorised according to their SPF,¹³ UVA protection and stability.^{14 15} SPF is defined as the amount of UV radiation (dose) required to produce minimal erythemal effects on protected skin relative to that of unprotected skin. SPF is generally, mainly a measure of UVB exposure as opposed to UVA exposure as it is weighted to the erythema (sunburn) spectrum. A sunscreen with an SPF of 15, properly applied and reapplied (defined as 2 mg/cm² of sun-exposed skin), will only allow 7% of erythemally effective energy to reach the skin, reducing to 3% for an SPF of 30 and 2% for an SPF of 50.¹⁶ Erythema is a common skin condition that causes redness or discolouration of the skin or mucous membranes. As the physiology of the ocular surface cells is not directly related to local blood flow, the SPF of ophthalmic products should be based on the action spectra for eye damage.

Establishing a CL-applicable SPF (CL-SPF) offers an opportunity to enhance communication on CL protection from the transmission of UV radiation. The aim of this study was to explore how a CL-SPF (defined as a measure of how much solar energy (UV) is required to produce damage on tissue protected by a contact lens relative to the amount of solar energy required to produce damage without a CL in situ) could be applied to CLs.

Materials and methods

The study was conducted in three parts. Part 1 determined UV transmittance and resulting CL-SPF through a range of CLs and spectacle lenses by adapting the method employed to determine the SPF of skin sunscreens. Part 2 explored solar angle effects and the influence of orientation between head and sun with two designs of sunglass to model different forms of eyewear. Part 3 evaluated UV damage to cornea and conjunctiva cells relative to the skin to refine CL-SPF values.

Part 1: contact lens and spectacle UV transmission

A system to measure the transmission of UV (280–380 nm) through soft hydrated CLs was developed (note: 400 nm was used as the upper limit as this is required by Cosmetics Europe (ISO 24444);¹⁷ a deuterium light source was selected (D2000, Ocean Optics, USA) to produce a high power output between 215 nm and 400 nm (spectral half-power bandwidth 210 to >1000 nm).

A Maya200-Pro Spectrometer (set to 0.25 nm wavelength interval measurements) with solarisation-resistant, fused silica, optical fibre patch cables (Ocean Optics, USA) was customised by replacing the standard detector window with a UV transparent variant. A system was designed to allow CLs to be inserted into the optical path between the light source and spectrophotometer optical transmission fibres (of 0.2 mm diameter) and held in place in direct contact between each of these aligned fibres so UV transmission could be determined through a 0.2 mm diameter segment of the contact lens. A precision three-axis stage PT3M (Thorlabs, USA) allowed centre and/or peripheral transmission to be measured.

Three separate spectra were required to determine the transmission profile: (1) a dark spectrum quantifying the noise produced by the spectrometer under non-illumination conditions, thus establishing a measurement baseline; (2) a light spectrum quantifying the level and wavelength spread of the UV source; and (3) a transmission spectrum of the lens measured.

Each of the three spectra was an average of three consecutive measurements through the centre of each −3.00 D CL over the entire measurement range (214–400 nm) at 0.25 nm intervals. In addition, measurements were captured every 1.5 mm from the centre to examine how CL-SPF varied over the diameter of the lens (as the UV blocker is in the CL material and therefore will vary with thickness). The lens was pressed between two fibre optics at each position to minimise any focusing effect of the lens optics.

This was repeated for 12 commercially available lenses at powers of +6.00 D, +3.00 D, −1.00 D, −3.00 D and (where available) −9.00 D.

Determining SPF involves measuring three factors: E(λ), the spectral irradiance of terrestrial sunlight; ελ, the erythema action spectrum; and PF(λ), the monochromatic protection factor. E(λ) and ελ were taken from Diffey and Robson¹⁸ (representing a sunlight irradiance latitude 40° north, solar zenith angle 20°, ozone layer thickness 30.5 mm) and the 1987 CIE (Commission on Illumination) approved erythema action spectrum (Groves et al).¹⁹ PF(λ) was determined from the measured transmission profiles. A MATLAB programme (MathWorks, USA) was developed to calculate the SPF as follows:

$${\displaystyle {\displaystyle CL-SPF=\frac{\sum { }_{290}^{400}\mathrm{E}\left(\lambda \right)\epsilon \lambda }{\sum { }_{290}^{400}\mathrm{E}\left(\lambda \right)\epsilon \lambda /\mathrm{P}\mathrm{F}\left(\lambda \right)}}}$$

Part 2: sun position and eyewear modelling

Ray tracing of the effect of solar angle (0°–90° in 10° steps) and head orientation relative to the sun (0°–90° in 10° steps) was undertaken with SolidWorks 2013 software (Dassault Systèmes, France). Sunglass and head models were modified from GrabCAD user-generated content. Sunglass models were https://grabcad.com/library/glasses-gfx001 (standard frame design, created by Gregory Felipe Guimarães) and https://grabcad.com/library/bmw_sunglasses-1 (wraparound design, created by Gustavo Mirra). The head model used was https://grabcad.com/library/head--4 (created by Sonu).

Part 3: action spectra profile of corneal and conjunctival cell damage

Established cell lines from ATCC—human conjunctival epithelial and corneal epithelial and PromoCell—normal adult human dermal fibroblasts were routinely cultured once the recommendations of roughly 10 000 cells per cm² were achieved for corneal and conjunctival epithelial cells and 3500 cells per cm² for dermal fibroblasts. Human conjunctival epithelial and human corneal epithelial were cultured in low glucose (1 g/L) Dulbecco’s Modified Eagle Medium containing 4 mM L-glutamine and 110 mg/L sodium pyruvate (Gibco, UK) supplemented with 10% v/v fetal bovine serum (FBS) and 100 µg/mL Penicillin/Streptomycin (Sigma-Aldrich, USA). Human dermal fibroblasts were cultured in Fibroblast Growth Medium 2 (PromoCell, UK) supplemented with the corresponding SupplementMix (PromoCell, UK). Cells were detached using 0.04% trypsin-EDTA (Sigma-Aldrich, USA), centrifuged at 300 g for 5 min (125 g for 5 min for corneal epithelial cells) and resuspended in the corresponding media above.

Corneal epithelial, conjunctival epithelial and dermal fibroblasts were seeded into 6-well plates and were incubated for 24–48 hours to achieve sufficient cell numbers. Media in wells was replaced with 1.5 mL fresh media to remove any dying cells prior to UV exposure. Wells were individually exposed to UV wavelengths from a UV light source at 10 nm intervals from 300 to 390 nm UV bandpass filters (Asahi Spectra USA, Torrance, California, USA) mounted in a filter wheel (Thorlabs, Newton, New Jersey, USA) above the cell seeded well plates. Variation in UV intensity through the filters from the light spectrum was adjusted by exposure time to produce a dose response damage curve and subsequently to apply equal doses of each wavelength to assess the relative damage profile across wavelengths. Cells were incubated at 37°C and 5% CO₂ and apoptosis was allowed to proceed for 24 or 48 hours.

Cells were exposed to UV at each of the 10 nm UV wavelength bands, and following a 24 or 48-hour incubation period to allow cell death to proceed, cells were stained to analyse levels of viability. Cells undergoing apoptosis expose phosphatidylserine (PS) irreversibly at the cell surface—signalling cellular damage and promoting cell removal in vivo.²⁰ In vitro, as cell death proceeds, membrane integrity becomes compromised, enabling the vital dye propidium iodide (PI) to enter the cells to stain nucleic acid. Together, annexin V (AxV) and PI enable sensitive detection of apoptotic cells (AxV+/PI–) and late apoptotic/necrotic (AxV+/PI+), and together these represent non-viable cells as a measure of cell damage. Cell death was assessed using AxV and PI staining following the manufacturer’s instructions and as previously described.²¹ Briefly, medium from wells containing non-viable cells was collected with viable adherent cells lifted from the wells with 0.04% trypsin-EDTA. Cells were centrifuged at 300 g for 5 min and resuspended in 500 µL AxV binding buffer (150 mM sodium chloride, 2.5 mM calcium chloride, 10 mM 4-(2-Hydroxyethyl)piperazine-1-ethane-sulfonic acid (HEPES)). 5 µL Annexin V- fluorescein isothiocyanate conjugated dye (eBioscience, UK) was added to each sample and incubated on ice for 5 min to incorporate and bind to surface PS exposed during apoptosis. Immediately prior to flow cytometry analysis using the BD Accuri C6 Plus Flow Cytometer, samples were stained with 5 µL PI, a fluorescent DNA binding dye. Cell death was assessed by comparison of AxV-FITC/PI staining relative to untreated cells. Untreated cells under an incandescent bulb acted as a negative control.

Following UV exposure and a 24 or 48-hour incubation, cell morphology was analysed using an EVOS FL Cell Imaging System (Thermo Fisher Scientific, UK) using the 10× objective lens. Photos taken from the EVOS acted as a qualitative measure of cell viability.

Results

Part 1: contact lens and spectacle lens transmission

The transmission spectra of the CLs and spectacle lenses are shown in figure 1.

Figure 1.

Transmission spectra for a range of commercial soft contact lenses (central measurement, −3.00 D power) and spectacle materials (no tint).

The CL-SPF values calculated from the spectra in figure 1 are summarised in table 1. The CL-SPF value maps over the surface of a range of CLs of different powers are shown in figure 2.

Table 1.

Calculated CL-SPF values from the UV transmission profiles for a range of commercial soft contact lenses (−3.00 D) and spectacle lens materials (plano)

Contact lens brand	Manufacturer	UV blocker	UVA % blocking (316–400 nm)	UVB % blocking (280–315 nm)	CL-SPF
AIR OPTIX AQUA	Alcon	No	15.5	21.5	1.2
DAILIES AquaComfortPlus		No	8	19	1.1
DAILIES TOTAL1		No	12.9	18.5	1.2
Biotrue ONEday	Bausch+Lomb	Class 2	84.7	96.5	20.8
PureVision2		No	12	33.8	1.4
SofLens Daily Disposable		No	2.7	8.6	1.0
Biofinity	CooperVision	No	3.9	1.8	1.0
Clariti 1 day		Class 2	71.9	96.2	12.3
MyDay		Class 2	83.8	95.1	17.2
Proclear 1 day		No	17.3	22.9	1.3
1 day ACUVUE MOIST	Johnson & Johnson Vision	Class 2	83.1	98.1	24.8
ACUVUE OASYS		Class 1	93.7	99.5	61.8
ACUVUE OASYS 1 day		Class 1	95.4	99.1	60.1
1 day ACUVUE TruEye		Class 1	95.1	99.6	66.2
ACUVUE VITA		Class 1	95.4	99.1	59.6
CR39	Spectacle lenses		87.4	99.4	46.2
Polycarbonate			99.6	99.4	156.9
Optical glass			7.7	65.9	1.8

CL-SPF, contact lens-Sun Protection Factor; UV, ultraviolet .

Figure 2.

Profile ‘maps’ of the contact lens-SPF that was measured through each annular zone of a range of commercial soft contact lenses at 1.5 mm intervals. SPF, Sun Protection Factor.

Part 2: sun position and eyewear modelling

Optical modelling showed maximum exposure without sunglasses occurred at a solar angle of 10° and head orientation of 10°, but dropped below 70% when the solar angle was >40° or head orientation was >20°. Light reached the ocular surface despite sunglass wear (figure 3). A standard frame allowed up to 52% of light to hit the ocular surface when the solar angle was high (80°−90°) with the head facing the sun; solar angle was less when the head-to-sun orientation was increased. Across all solar angles and head orientations, on average 76%–89% of the light was prevented from reaching the ocular surface, depending on the intensity of the tint (80%–20% transmission). With wraparound sunglasses, virtually no light reached the ocular surface around the frame (2%), although this increased by 428% when the frame was displaced 6 mm from the bridge of the nose.

Figure 3.

Ocular surface exposure to light rays with solar angle (from perpendicular to the face and head orientation (from aligned with the source) relative to the source (in degrees) with non-wraparound sunglasses.

Part 3: damage profile of corneal and conjunctival cells

To ascertain the effect of both UVA and UVB exposure on corneal and conjunctival epithelial cells compared with dermal cells, cells were exposed to a 10 nm wide range of wavelengths within the UVA and UVB spectra. This was repeated with fresh cells for 10 nm UV wavelength ranges across the UV spectrum (275–285, 285–295, 295–305, 305–315, 315–325, 325–335, 335–345, 345–355, 355–365, 365–375, 375–285, 385–395 nm). All three cell types were exposed to a range of UV wavelengths, spanning both the UVA to visible (320–400 nm) and UVB (280–320 nm) spectra. To assess damage to cells post-UV exposure, cells were analysed using multicolour flow cytometry. AxV staining was used to detect exposed PS and membrane integrity was assessed using the vital dye PI for staining of nucleic acids when membrane integrity is lost. Relative damage to tissue (figure 4) is defined as the sum of UV-exposed cells that were apoptotic (AxV+/PI−) or necrotic (AxV+/PI+) relative to an untreated healthy control. Cell viability was found to be >70% pre-exposure for all cell types. Post-exposure, however, cell viability was compromised as a result of cell damage throughout the UVB and a significant portion of the UVA spectrum (figure 4).

Figure 4.

Action damage spectra profile over the ultraviolet radiation range of corneal and conjunctival epithelial cells, compared with skin cells. Each wavelength is the midpoint of the ultraviolet 10 nm band, so 280 nm=a 275–285 nm wavelength band.

Both skin tissue and corneal and conjunctival cells exhibited a similar relative damage profile when exposed to wavelengths within the UVB spectrum. Unlike skin tissue, however, which experiences a low relative damage profile, corneal and conjunctival cells showed a greater relative damage profile across the UVA spectral range, in particular between 320 nm and 400 nm, the bulk of the UVA spectrum.

Discussion

CLs vary in ocular UV transmission and CL-SPF as assessed by in vitro methodology developed for skin sunscreen. Transmissions measured were similar to those previously reported.^22–24 Differences may be accounted for by the area of lens measured (0.4 mm² compared with 6 mm²), differences in lens power measured and range of UV considered (up to 400 nm with the COLIPA method compared with 380 nm generally). The present study allowed measurements at 1.5 mm intervals over the diameter of the CLs, enabling CL-SPF maps to be produced for five lens powers. Unlike Lin et al,²⁵ expected differences with lens thickness were shown, especially noticeable in Class 2 UV-blocking CLs, but demonstrated high levels of blocking at all positions across the Class 1 UV-blocking CLs. Blocking transmission to the peripheral cornea, conjunctiva and crystalline lens is important because of the location of common ophthalmohelioses.⁹

SPF is widely recognised by consumers and can be communicated when reminding them of the need for comprehensive sun protection. Clinically, the SPF relates to the additional multiples of time an individual can remain exposed to UV without the effects of burning. An individual with fair skin that burns naturally in 10 min exposure to UV with skin-SPF 15 (appropriately applied) could be exposed to UV for 150 min without burning. The same logic could be applied to the conjunctiva and crystalline lens, although the UV damage to these tissues has not been well quantified; whether this depends on individual factors such as age and pigmentation has yet to be adequately confirmed.

Class 1 and 2 UV-blocking CLs also have UVA blocking, which is minimally accounted for when weighting by a UV cell damage spectra. In addition, CLs are a fixed thickness and are not absorbed by the underlying tissue, or rubbed off by contact or liquids, as with sunscreens. Hence their effectiveness is not reliant on these compliance factors.²⁶ In addition, CLs are worn most of the time, regardless of weather conditions; hence UV-blocking CLs may protect the ocular surface from UV for longer periods than sunglasses, which tend to be worn only when it is sunny, despite significant UV penetration through clouds.

Although the eyes have natural sun protection (from being inset in the orbit and from the eye brows) which can be enhanced by spectacles or sunglasses, these have an SPF of 2–25 which will be reduced by~8%–26% depending on the solar angle, head orientation, lens material and tint and frame. CL-SPF does not account for the sun’s intensity, which is generally greater the higher the solar angle, but will vary with geographical location.¹⁰ The number of variables in modelling the effectiveness of sunglasses against UV is vast, including the frame component shapes, lens material and tinting, properties of any coatings, back vertex distance, orbital anatomy, brow dimensions and environmental reflections. Hence the best modelling can achieve is to indicate that, even if worn whenever there is UV (including cloudy days), sunglasses will not prevent all UV reaching the ocular surface. Wrap-around sunglasses with plastic lenses and a low transmission will, however, give protection to the conjunctival tissue beyond the area protected by UV-blocking CLs, even those with a high CL-SPF, and therefore should be advocated as part of comprehensive sun protection, together with a wide-brimmed hat to further protect the ocular adnexa.²⁷

Citek¹¹ evaluated reflectance from spectacle lenses, with and without anti-reflection coatings or tints and showed that antireflection coatings could potentially reflect damaging wavelengths into the eye. This author and others developed an E-SPF, based on physical measurements of spectacle lens material transmission and reflection weighted for spectral distribution for solar irradiation between 280 nm and 380 nm, but not for an erythemal or equivalent ocular cell damage profile. Despite a similar range of values to the ocular tissue-corrected SPF in the present study, E-SPF does not account for solar angle, head orientation relative to the sun or potentially magnified effects of peripheral light.^{11 12} Other authors have observed that a standardised, reliable and comprehensive label for UV eye protection, for consumers and professionals, is currently lacking and proposed a strategy for establishing biological relevance for the E-SPF.²⁸ While SPF has a very specific application and meaning in international standards and is more than a simple ratio, rather being considered an in vivo method of measurements and calculation, it is a regularly encountered term by consumers and useful for communication regarding UV protection.

A traditional SPF level is based on the erythemal spectrum of skin cells which is principally UVB limited. Hence for CL protection, the action damage spectra of corneal and conjunctival cells should be substituted. The present study appears to be the first to assess true cellular damage across a wide range of narrow UV radiation bands. The findings demonstrate that damage to the conjunctiva is greater than that to the cornea and, compared with the traditional skin cell erythemal profile, UVA has a much more significant effect on conjunctival cells than previously thought. UV radiation is known to cause oxidative damage by generating oxygen-derived free radicals, especially in the mitochondria. This causes damage to biomolecules such as proteins, lipids and nucleic acids, leading to cell damage and eventual cell death. These findings imply even greater importance of UV filtering by CLs in the form of a higher CL-SPF. It is important to note that a standard soft contact lens does not cover the entire conjunctiva, and so any protection afforded by UV-blocking properties is limited to the limbal area, cornea and internal ocular structures. A rigid corneal lens will only protect some of the central cornea and reduce UV entering the pupil. However, larger ‘specialist’ soft CLs and scleral lenses will provide ocular surface protection over a wider area. A similar limitation also applies to the SPF quoted for sunscreen lotions which is based on a thick layer (2 mg/cm² of sun-exposed skin) and regular reapplication,¹⁶ and the SPF of clothes, which similar to CLs, will only protect the area that they cover. A recent study found that blocking UV transmission through asoft contact lens for at least 5 years was beneficial in maintaining the eyes' ability to focus, suggesting that presbyopia may be delayed in long-term UV-blocking soft CL wearers; soft contact lenses also provide protection to the critical limbal region.²⁹

To be comparable with the approach taken to generate SPF for sunscreens, it is proposed that the original skin-based SPF bands are retained, which separates CLs into three categories:

CL-SPF with no UV blocker=1.0–2.0—equivalent to using no sunscreen.

CL-SPF equivalent to Class 2 UV blocker=12.2–24.8—equivalent to sunscreen SPF 15, taken as the level used by many adults in moderately sunny conditions.

CL-SPF equivalent to Class 1 UV blocker=59.6–66.2—equivalent to sunscreen 50+ (adopted as a labelling restriction maximum by the European Union and Australia, as well as being proposed by the Food and Drug Administration), generally applied to children and people with fair skin.

In conclusion, this study provides a CL-SPF quantification technique for CLs which differentiates between no UV blocker, Class 1 and Class 2 UV blocking and uses classifications known and accepted by consumers. While ocular cells seem to have a similar (relative damage profile to the skin, they seem to be more susceptible to UV damage, making a higher CL-SPF even more critical. Further investigations are needed to determine whether there are potential differences between a cellular culture model compared with in vivo measurement. However, the present study shows that CL-SPF is a viable metric to communicate the protection from the transmission of UV that some CLs offer wearers.

Data availability statement

Data are available upon reasonable request.

Ethics statements

Patient consent for publication

Not applicable.

Ethics approval

The cells used in this study were sourced from PromoCell who state 'the isolation of human cell cultures is derived from donors who have signed an informed consent form (this being done by the donor himself, an authorized agent, or a legal agent) which outlines in detail the purpose of the donation and the procedure for processing the tissue'.