A program to analyse optical coherence tomography images of the ciliary muscle

Purpose: To describe and validate bespoke software designed to extract morphometric data from ciliary muscle Visante Anterior Segment Optical Coherence Tomography (AS-OCT) images. Method: Initially, to ensure the software was capable of appropriately applying tiered refractive index corrections and accurately measuring orthogonal and oblique parameters, 5 sets of custom-made rigid gas-permeable lenses aligned to simulate the sclera and ciliary muscle were imaged by the Visante AS-OCT and were analysed by the software. Human temporal ciliary muscle data from 50 participants extracted via the internal Visante AS-OCT caliper method and the software were compared. The repeatability of the software was also investigated by imaging the temporal ciliary muscle of 10 participants on 2 occasions. Results: The mean difference between the software and the absolute thickness measurements of the rigid gas-permeable lenses were not statistically significantly different from 0 (t = -1.458, p = 0.151). Good correspondence was observed between human ciliary muscle measurements obtained by the software and the internal Visante AS-OCT calipers (maximum thickness t = -0.864, p = 0.392, total length t = 0.860, p = 0.394). The software extracted highly repeatable ciliary muscle measurements (variability ≤6% of mean value). Conclusion: The bespoke software is capable of extracting accurate and repeatable ciliary muscle measurements and is suitable for analysing large data sets.

International flavours

It is an Olympic year and we have just witnessed the fantastic games hosted by Rio de Janeiro. Well done to team USA for winning the most medals overall but also well done to so many other nations and individuals who performed so well or were ambassadors in other ways. Teenage swimmer Yusra Mardini who swam for the refugee team and South Africa's Wayde van Niekerk who broke the longstanding 400 m record of Michael Johnson that has stood since 1999. Of course, we must mention sprinter Usain Bolt and swimmer Michael Phelps, who have now transcended superstar status and entered a new level of icon. My personal highlight was the sportsmanship witnessed in the 5000 m when American Abbey D’Agostino was accidentally felled by New Zealand runner Nikki Hamblin. D’Agostino helped Hamblin back to her feet but slumped to the track after realising her own injury. Hamblin helped her up and stayed with her so that both completed the race. The International Olympic Committee has awarded both with the prestigious Pierre de Coubertin award, also known as the International Fair Play Trophy. Fair play is of paramount importance in publishing in peer-reviewed papers. At CLAE we try and maintain, as do other journals, this by ensuring double blind peer review and allowing authors to select the most appropriate handling editor for their submission. Our handling editors are placed across the world (2 in Europe, 1 in the Americas, 1 in Australia and 1 in Asia) and part of their role is to encourage submissions from their region. Over the last decade we certainly have seen more and more papers from places that haven’t previously published in CLAE. In this issue of CLAE we have a true international blend of papers. We have papers from authors from the UK, USA, Iran, Jordan, France, Poland, Turkey, Nigeria, France, Spain and Brazil. I think it's a testament to the continued success of the journal that we are attracting new writers from so many parts of the world and retain papers from more established authors and research centres. We do continue to attract many weaker papers that are rejected early in the review process. Often these will be unexceptional case reports or papers describing a surgical technique. Case reports are published but only those that offer something original and especially those with interesting photographs. In this issue you will see Professor James Wolffsohn (UK) has an interesting paper around a lot of the focus of his recent research activity into clinical evaluation of methods of correcting presbyopia. In this paper he highlights predictors to aid success of presbyopic contact lenses. If you have been involved in any clinical work or research in the field of dry eye disease then you will know well the CLDEQ (Contact Lens Dry Eye Questionnaire) devised by Robin Chalmers and her colleagues (USA). This issue of CLAE details the latest research using the CLDEQ-8 (the 8 item version of the CLDEQ). The Shahroud Eye Cohort Study has produced many papers already and in this issue we see Fotouhi Akbar (Iran) looking at changes in central and peripheral corneal thickness over a five year period. These days we use a lot of new instrumentation, such as optical low-coherence reflectometry. In this issue Emre Güler (Turkey) compares that to a new optical biometry unit. Dry eye is more common and in this issue we see a study by Oluyemi Fasina (Nigeria) to investigate the disease in adults in South-West Nigeria. The TearLab™ is now commonly used to investigate osmolarity and Dorota Szczesna-Iskander (Poland) looks at measurement variability of this device. Following the theme of dry eyes and tear testing Renaud Laballe (France) looks at the use of scleral lenses as a reservoir-based ocular therapeutic system. In this issue we have a couple of papers looking at different aspects of keratoconus. Magdalena Popiela (UK) looks at demographics of older keratoconic patients in Wales, Faik Orucoglu (Turkey) reports a novel scoring system for distinguishing keratoconus from normal eyes, Gonzalo Carracedo (Spain) reports the effect of rigid gas permeable lens wear on dry eye in keratoconus and Hatice Nur Colak (Turkey) compares topographic and aberrations in keratoconus. Other interesting papers you will find are Mera Haddad (Jordan) investigates contact lens prescribing in Jordan, Camilla Fraga Amaral (Brazil) offers a report on the use of ocular prosthetics, Naveed Ahmed Khan (Malaysia) reports of the use of dimethyl sulfoxide in contact lens disinfectant and Michael Killpartrick (UK) offers a short piece with some useful advice on contamination risk factors that may occur from the posterior surface of disposable lenses. So for this issue I would say that the Gold Medal for biggest contribution in terms of papers has to go to Turkey. I could have awarded it to the UK too, but Turkey has three full papers and the UK has two plus one short communication. Turkey is also one of the countries that has shown the largest increase in submissions over the last decade. Finally, welcome aboard to our newest Editorial Board Member Nicole Carnt from Australia. Nicole has been an active researcher for many years and acted as a reviewer for CLAE many times in the past. We look forward to working with you.

Ultraviolet photodetection of flexible ZnO nanowire sheets in polydimethylsiloxane polymer

ZnO nanowires are normally exposed to an oxygen atmosphere to achieve high performance in UV photodetection. In this work we present results on a UV photodetector fabricated using a flexible ZnO nanowire sheet embedded in polydimethylsiloxane (PDMS), a gas-permeable polymer, showing reproducible UV photoresponse and enhanced photoconduction. PDMS coating results in a reduced response speed compared to that of a ZnO nanowire film in air. The rising speed is slightly reduced, while the decay time is prolonged by about a factor of four. We conclude that oxygen molecules diffusing in PDMS are responsible for the UV photoresponse

Packed bed bioreactor for the isolation and expansion of placental-derived Mesenchymal Stromal Cells

Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs.

Optical sensors for carbon dioxide: an overview of sensing strategies past and present

The evolution of the optical sensor for CO2 over the past two decades is outlined and illustrated through examples of luminescent-based sensors. The basic principles and design of the early 'wet covered' type sensor, in which a pH sensitive dye in an aqueous buffer is covered by a gas permeable, ion impermeable, membrane, are outlined. The gradual move from the 'wet covered' types of CO2 optical sensor to 'solid-water droplet' type sensors and then onto 'solid' sensors is charted. The basic design and principles of operation of the modern 'solid' optical sensor for P-CO2 is covered in some detail. Other sensing strategies outside the simple use of pH-sensitive dyes are also considered, most notably those based on luminescence lifetime measurements.

Use of contact lenses in patients with filtering blebs

Purpose: The purpose of this study was to describe the outcome of patients with filtering blebs who were fit with contact lenses. Methods: We retrospectively studied patients with filtering blebs secondary to glaucoma or cataract surgery who were fit with contact lenses. Eight eyes from seven patients were identified. Results: Five patients (six eyes) were fit with gas permeable contact lenses and two patients (two eyes) were fit with soft contact lenses. Successful fits were achieved in all patients. No complications were observed after a mean follow-up of 64.6±28.5 months. Conclusions: No significant complications were recorded in our series of patients with filtering blebs who were fit with contact lenses. We think that when indications for fitting contact lenses are justified, patients with filtering blebs are acceptable candidates for contact lens use. However, adequate selection of cases, careful contact lens fitting, patient education, and close follow-up are necessary.

Contact lens use after contact lens-associated infectious ulcers

Purpose: The purpose of this study was to evaluate the outcome of patients with healed moderate to severe contact lens-associated corneal infectious ulcers who were re-fit with contact lenses. Methods: We retrospectively studied patients who were fit with contact lenses on our service and who had had moderate to severe corneal infectious ulcers associated with previous contact lens use. Six patients were included in the study. Results: Gas permeable contact lenses were fit in five patients, and a soft contact lens was fit in one patient. Successful fit was achieved in all cases and visual acuities were equal to or better than 20/30 in all patients. No significant complications were observed after a mean follow-up of 23 months (range, 6-45 months). Conclusion: In this small series of patients with a history of moderate to severe contact-lens related infectious keratitis, no complications were observed after contact lenses were refit. Contact lens wear in patients with a history of infectious keratitis may be safe and useful in order to achieve visual rehabilitation.

Continuous-flow processing of gaseous ammonia using a Teflon AF-2400 tube-in-tube reactor: synthesis of thioureas and in-line titrations

A simple tube-in-tube reactor based on the gas-permeable membrane Teflon AF-2400 was used in the continuous flow reaction of gaseous ammonia with isothiocyanates and one isocyanate. A colourimetric in-line titration technique is also reported as a simple method to quantify the amount of ammonia taken up by the solvent in the system.

Properties of seawater (partial pressure of carbon dioxide (pCO2)) from a ProOceanus CO2-Pro sensor mounted on the continuous surface water sampling system during the Tara Oceans expedition 2009-2013

The Tara Oceans Expedition (2009-2013) sampled the world oceans on board a 36 m long schooner, collecting environmental data and organisms from viruses to planktonic metazoans for later analyses using modern sequencing and state-of-the-art imaging technologies. Tara Oceans Data are particularly suited to study the genetic, morphological and functional diversity of plankton. The present data set provides continuous measurements of partial pressure of carbon dioxide (pCO2), using a ProOceanus CO2-Pro instrument mounted on the flowthrough system. This automatic sensor is fitted with an equilibrator made of gas permeable silicone membrane and an internal detection loop with a non-dispersive infrared detector of PPSystems SBA-4 CO2 analyzer. A zero-CO2 baseline is provided for the subsequent measurements circulating the internal gas through a CO2 absorption chamber containing soda lime or Ascarite. The frequency of this automatic zero point calibration was set to be 24 hours. All data recorded during zeroing processes were discarded with the 15-minute data after each calibration. The output of CO2-Pro is the mole fraction of CO2 in the measured water and the pCO2 is obtained using the measured total pressure of the internal wet gas. The fugacity of CO2 (fCO2) in the surface seawater, whose difference with the atmospheric CO2 fugacity is proportional to the air-sea CO2 fluxes, is obtained by correcting the pCO2 for non-ideal CO2 gas concentration according to Weiss (1974). The fCO2 computed using CO2-Pro measurements was corrected to the sea surface condition by considering the temperature effect on fCO2 (Takahashi et al., 1993). The surface seawater observations that were initially estimated with a 15 seconds frequency were averaged every 5-min cycle. The performance of CO2-Pro was adjusted by comparing the sensor outputs against the thermodynamic carbonate calculation of pCO2 using the carbonic system constants of Millero et al. (2006) from the determinations of total inorganic carbon (CT ) and total alkalinity (AT ) in discrete samples collected at sea surface. AT was determined using an automated open cell potentiometric titration (Haraldsson et al. 1997). CT was determined with an automated coulometric titration (Johnson et al. 1985; 1987), using the MIDSOMMA system (Mintrop, 2005). fCO2 data are flagged according to the WOCE guidelines following Pierrot et al. (2009) identifying recommended values and questionable measurements giving additional information about the reasons of the questionability.

Prolonged production of NADPH oxidase-corrected granulocytes after gene therapy of chronic granulomatous disease

Little is known about the potential for engraftment of autologous hematopoietic stem cells in human adults not subjected to myeloablative conditioning regimens. Five adult patients with the p47phox deficiency form of chronic granulomatous disease received intravenous infusions of autologous CD34+ peripheral blood stem cells (PBSCs) that had been transduced ex vivo with a recombinant retrovirus encoding normal p47phox. Although marrow conditioning was not given, functionally corrected granulocytes were detectable in peripheral blood of all five patients. Peak correction occurred 3–6 weeks after infusion and ranged from 0.004 to 0.05% of total peripheral blood granulocytes. Corrected cells were detectable for as long as 6 months after infusion in some individuals. Thus, prolonged engraftment of autologous PBSCs and continued expression of the transduced gene can occur in adults without conditioning. This trial also piloted the use of animal protein-free medium and a blood-bank-compatible closed system of gas-permeable plastic containers for culture and transduction of the PBSCs. These features enhance the safety of PBSCs directed gene therapy.

Specialist contact lens fitting

What is meant by the term ‘specialist contact lens fitting’? Or put another way, what would be considered non-specialist contact lens fitting? Is there such a thing as routine contact lens fitting? Soft or silicone hydrogel fitting for daily wear would probably be considered as routine contact lens fitting, but would extended or flexible wear remain in the same category or would they be considered a specialist fit? Different eras will classify different products as being ‘specialist’. Certainly twenty years ago soft toric contact lenses were considered as being speciality lenses but today would be thought of as routine lenses. Conversely, gas permeable lenses were thought of as mainstream twenty years ago but now are considered as speciality lenses. Although this would not be the same globally, as in some countries (such as Netherlands, France and Japan) gas permeable lens fitting remains popular and is not on the decline as in other countries (Canada, Australia and Sweden) [1]. Bandage soft lenses applied after surface laser refractive procedures would be considered as therapeutic lenses but in reality they are just plano thin hydrogel lenses worn constantly for 3–4 days to allow the underlying epithelium to convalesce and are then removed [2]. Some patients find that wearing hydrogel lenses during periods when they suffer from seasonal allergies actually improves their ocular comfort as the contact lens acts as a barrier to the allergen [3] and [4]. Scleral lenses have long been considered speciality lenses, apart from a time when they were the only lenses available but at that time all contact lens work would have been considered speciality practice! Nowadays we see the advent of mini-scleral designs and we see large diameter gas permeable lenses too. It is possible that these lenses increase the popularity of gas permeable lenses again and they become more main stream. So it would seem that the lines between routine and speciality contact lens fitting are not clear. Whether a lens is classed a specialist fit or not would depend on the lens type, why it was fitted, where in the world the fitting was being done and even the era in which it was fitted. This begs the question as to what would be considered entry level knowledge in contact lens fitting. This may not be an issue for most BCLA members or CLAE readers but certainly would be for bodies such as the College of Optometrists (UK) or the Association of British Dispensing Opticians when they are planning the final registration examinations for budding practitioners or when planning the level of higher level qualifications such as College Certificates or Diplomas. Similarly for training institutions when they are planning their course content. This becomes even trickier when trying to devise a qualification that spans across many countries, like the European Diploma in Optometry and Optics. How do we know if the training and examination level is correct? One way would be to analyse things when they go wrong and if patterns of malpractice are seen then maybe that could be used as an indicator to more training being needed. There were 162 Fitness to Practice Hearing at the General Optical Council between 2001 and 2010. Forty-seven of these were clinically related case, 39 fraud related, and 76 others. Of the clinical ones only 3 were contact lens related. So it would appear that as whole, in the profession, contact lens clinical skills are not being questioned too often (although it seems a few of us can’t keep our hands out the cookie jar!).

Changes at the BCLA

As we welcome 2014 we say goodbye to 2013 and I must start with an apology to authors who have submitted papers to CLAE and seen a delay in either the review process or the hard copy publication of their proofed article. The delays were caused by a major hike in the number of submissions to the journal in 2012 that increased further in 2013. In the 12 months leading to the end of October 2011 we had 94 new paper submissions, and for the same period to the end of 2012 the journal had 116 new papers. In 2012 we were awarded an impact factor for the first time and following that the next 12 month period to the end of October 2013 saw a massive increase in submissions with 171 new manuscripts being submitted. This is nearly twice as many papers as 2 years ago and 3 times as many as when I took over as Editor-in-Chief. In addition to this the UK academics will know that 2014 is a REF year (Research Excellence Framework) where universities are judged on their research and one of the major components of this measure remains to be published papers so there is a push to publishing before the REF deadline for counting. The rejection rate at CLAE has gone up too and currently is around 50% (more than double the rejection rate when I took over as Editor-in-Chief). At CLAE the number of pages that we publish each year has remained the same since 2007. When compiling issue 1 for 2014 I chose the papers to be included from the papers that were proofed and ready to go and there were around 200 proofed pages ready, which is enough to fill 3½ issues! At present Elsevier and the BCLA are preparing to increase the number the pages published per issue so that we can clear some of this backlog and remain up to date with the papers published in CLAE. I should add that on line publishing of papers is still available and there may have been review delays but there are no publishing online so authors can still get an epub on line final version of their paper with a DOI (digital object identifier) number enabling the paper to be cited. There are two awards that were made in 2013 that I would like to make special mention of. One was for my good friend Jan Bergmanson, who was awarded an honorary life fellowship of the College of Optometrists. Jan has served on the editorial board of CLAE for many years and in 2013 also celebrated 30 years of his annual ‘Texan Corneal and contact lens meeting’. The other award I wish to mention is Judith Morris, who was the BCLA Gold Medal Award winner in 2013. Judith has had many roles in her career and worked at Moorfields Eye Hospital, the Institute of Optometry and currently at City University. She has been the Europe Middle East and Africa President of IACLE (International Association of Contact Lens Educators) for many years and I think I am correct in saying that Judith is the only person who was President of both the BCLA (1983) and a few years later she was the President College of Optometrists (1989). Judith was also instrumental in introducing Vivien Freeman to the BCLA as they had been friends and Judith suggested that Vivien apply for an administrative job at the BCLA. Fast forward 29 years and in December 2013 Vivien stepped down as Secretary General of the BCLA. I would like to offer my own personal thanks to Vivien for her support of CLAE and of me over the years. The BCLA will not be the same and I wish you well in your future plans. But 2014 brings in a new position to the BCLA – Cheryl Donnelly has been given the new role of Chief Executive Officer. Cheryl was President of the BCLA in 2000 and has previously served on council. I look forward to working with Cheryl and envisage a bright future for the BCLA and CLAE. In this issue we have some great papers including some from authors who have not published with CLAE before. There is a nice paper on contact lens compliance in Nepal which brings home some familiar messages from an emerging market. A paper on how corneal curvature is affected by the use of hydrogel lenses is useful when advising patients how long they should leave their contact lenses out for to avoid seeing changes in refraction or curvature. This is useful information when refracting these patients or pre-laser surgery. There is a useful paper offering tips on fitting bitoric gas permeable lenses post corneal graft and a paper detailing surgery to implant piggyback multifocal intraocular lenses. One fact that I noted from the selection of papers in the current issue is where they were from. In this issue none of the corresponding authors are from the United Kingdom. There are two papers each from the United States, Spain and Iran, and one each from the Netherlands, Ireland, Republic of Korea, Australia and Hong Kong. This is an obvious reflection of the widening interest in CLAE and the BCLA and indicates the new research groups emerging in the field.

Extruded colour-based plastic film for the measurement of dissolved CO2

Dissolved CO2 measurements are usually made using a Severinghaus electrode, which is bulky and can suffer from electrical interference. In contrast, optical sensors for gaseous CO2, whilst not suffering these problems, are mainly used for making gaseous (not dissolved) CO2 measurements, due to dye leaching and protonation, especially at high ionic strengths (>0.01 M) and acidity (<pH 4). This is usually prevented by coating the sensor with a gas-permeable, but ion-impermeable, membrane (GPM). Herein, we introduce a highly sensitive, colourimetric-based, plastic film sensor for the measurement of both gaseous and dissolved CO2, in which a pH-sensitive dye, thymol blue (TB) is coated onto particles of hydrophilic silica to create a CO2-sensitive, TB-based pigment, which is then extruded into low density polyethylene (LDPE) to create a GPM-free, i.e. naked, TB plastic sensor film for gaseous and dissolved CO2 measurements. When used for making dissolved CO2 measurements, the hydrophobic nature of the LDPE renders the film: (i) indifferent to ionic strength, (ii) highly resistant to acid attack and (iii) stable when stored under ambient (dark) conditions for >8 months, with no loss of colour or function. Here, the performance of the TB plastic film is primarily assessed as a dissolved CO2 sensor in highly saline (3.5 wt%) water. The TB film is blue in the absence of CO2 and yellow in its presence, exhibiting 50% transition in its colour at ca. 0.18% CO2. This new type of CO2 sensor has great potential in the monitoring of CO2 levels in the hydrosphere, as well as elsewhere, e.g. food packaging and possibly patient monitoring.

Self-sealing of thermal fatigue and mechanical damage in fiber-reinforced composite materials

Fiber reinforced composite tanks provide a promising method of storage for liquid oxygen and hydrogen for aerospace applications. The inherent thermal fatigue of these vessels leads to the formation of microcracks, which allow gas phase leakage across the tank walls. In this dissertation, self-healing functionality is imparted to a structural composite to effectively seal microcracks induced by both mechanical and thermal loading cycles. Two different microencapsulated healing chemistries are investigated in woven glass fiber/epoxy and uni-weave carbon fiber/epoxy composites. Self-healing of mechanically induced damage was first studied in a room temperature cured plain weave E-glass/epoxy composite with encapsulated dicyclopentadiene (DCPD) monomer and wax protected Grubbs' catalyst healing components. A controlled amount of microcracking was introduced through cyclic indentation of opposing surfaces of the composite. The resulting damage zone was proportional to the indentation load. Healing was assessed through the use of a pressure cell apparatus to detect nitrogen flow through the thickness direction of the damaged composite. Successful healing resulted in a perfect seal, with no measurable gas flow. The effect of DCPD microcapsule size (51 um and 18 um) and concentration (0 - 12.2 wt%) on the self-sealing ability was investigated. Composite specimens with 6.5 wt% 51 um capsules sealed 67% of the time, compared to 13% for the control panels without healing components. A thermally stable, dual microcapsule healing chemistry comprised of silanol terminated poly(dimethyl siloxane) plus a crosslinking agent and a tin catalyst was employed to allow higher composite processing temperatures. The microcapsules were incorporated into a satin weave E-glass fiber/epoxy composite processed at 120C to yield a glass transition temperature of 127C. Self-sealing ability after mechanical damage was assessed for different microcapsule sizes (25 um and 42 um) and concentrations (0 - 11 vol%). Incorporating 9 vol% 42 um capsules or 11 vol% 25 um capsules into the composite matrix leads to 100% of the samples sealing. The effect of microcapsule concentration on the short beam strength, storage modulus, and glass transition temperature of the composite specimens was also investigated. The thermally stable tin catalyzed poly(dimethyl siloxane) healing chemistry was then integrated into a [0/90]s uniweave carbon fiber/epoxy composite. Thermal cycling (-196C to 35C) of these specimens lead to the formation of microcracks, over time, formed a percolating crack network from one side of the composite to the other, resulting in a gas permeable specimen. Crack damage accumulation and sample permeability was monitored with number of cycles for both self-healing and traditional non-healing composites. Crack accumulation occurred at a similar rate for all sample types tested. A 63% increase in lifetime extension was achieved for the self-healing specimens over traditional non-healing composites.

Theory of Gas Hydrates: Effect of the Approximation of Rigid Water Lattice

One of the assumptions of the van der Waals and Platteeuw theory for gas hydrates is that the host water lattice is rigid and not distorted by the presence of guest molecules. In this work, we study the effect of this approximation on the triple-point lines of the gas hydrates. We calculate the triple-point lines of methane and ethane hydrates via Monte Carlo molecular simulations and compare the simulation results with the predictions of van der Waals and Platteeuw theory. Our study shows that even if the exact intermolecular potential between the guest molecules and water is known, the dissociation temperatures predicted by the theory are significantly higher. This has serious implications to the modeling of gas hydrate thermodynamics, and in spite of the several impressive efforts made toward obtaining an accurate description of intermolecular interactions in gas hydrates, the theory will suffer from the problem of robustness if the issue of movement of water molecules is not adequately addressed.