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Stop codon miscoding events give rise to longer proteins, which may alter the protein’s function and thereby generate short-lasting phenotypic variability from a single gene.In order to systematically assess the frequency and origin of stop codon miscoding events, we designed a library of reporters. We introduced premature stop codons into mScarlet that enabled high-throughput quantification of protein synthesis termination errors inE.coliusing fluorescent microscopy. We found that under stress conditions, stop codon miscoding may occur with a rate as high as 80%, depending on the nucleotide context, suggesting that evolution frequently samples stop codon miscoding events. The analysis of selected reporters by mass spectrometry and RNA-seq showed that not only translation but also transcription errors contribute to stop codon miscoding. The RNA polymerase is more likely to misincorporate a nucleotide at premature stop codons. Proteome-wide detection of stop codon miscoding by mass spectrometry revealed that temperature regulates the expression of cryptic sequences generated by stop codon miscoding inE.coli.Overall, our findings suggest that the environment influences the accuracy of protein production, which increases protein heterogeneity when the organisms need to adapt to new conditions.

Cells exposed to starvation have to adjust their metabolism to conserve energy and protect themselves. Protein synthesis is one of the major energy-consuming processes and as such has to be tightly controlled. Many mechanistic details about how starved cells regulate the process of protein synthesis are still unknown. Here, we report that the essential translation initiation factor eIF2B forms filaments in starved budding yeast cells. We demonstrate that filamentation is triggered by starvation-induced acidification of the cytosol, which is caused by an influx of protons from the extracellular environment. We show that filament assembly by eIF2B is necessary for rapid and efficient downregulation of translation. Importantly, this mechanism does not require the kinase Gcn2. Furthermore, analysis of site-specific variants suggests that eIF2B assembly results in enzymatically inactive filaments that promote stress survival and fast recovery of cells from starvation. We propose that translation regulation through filament assembly is an efficient mechanism that allows yeast cells to adapt to fluctuating environments., Summary: Filament formation by eIF2B is an adaptive mechanism that allows for efficient shutdown of translation in starved yeast cells.

How cells adapt to varying environmental conditions is largely unknown. Here, we show that, in budding yeast, the RNA-binding and stress granule protein Pub1 has an intrinsic property to form condensates upon starvation or heat stress and that condensate formation is associated with cell-cycle arrest. Release from arrest coincides with condensate dissolution, which takes minutes (starvation) or hours (heat shock). In vitro reconstitution reveals that the different dissolution rates of starvation- and heat-induced condensates are due to their different material properties: starvation-induced Pub1 condensates form by liquid-liquid demixing and subsequently convert into reversible gel-like particles; heat-induced condensates are more solid-like and require chaperones for disaggregation. Our data suggest that different physiological stresses, as well as stress durations and intensities, induce condensates with distinct physical properties and thereby define different modes of stress adaptation and rates of recovery., Cell Reports, 23 (11), ISSN:2666-3864, ISSN:2211-1247

Summary Cells sense elevated temperatures and mount an adaptive heat shock response that involves changes in gene expression, but the underlying mechanisms, particularly on the level of translation, remain unknown. Here we report that, in budding yeast, the essential translation initiation factor Ded1p undergoes heat-induced phase separation into gel-like condensates. Using ribosome profiling and an in vitro translation assay, we reveal that condensate formation inactivates Ded1p and represses translation of housekeeping mRNAs while promoting translation of stress mRNAs. Testing a variant of Ded1p with altered phase behavior as well as Ded1p homologs from diverse species, we demonstrate that Ded1p condensation is adaptive and fine-tuned to the maximum growth temperature of the respective organism. We conclude that Ded1p condensation is an integral part of an extended heat shock response that selectively represses translation of housekeeping mRNAs to promote survival under conditions of severe heat stress.

Stressed cells shut down translation, release mRNA molecules from polysomes, and form stress granules (SGs) via a network of interactions that involve G3BP. Here we focus on the mechanistic underpinnings of SG assembly. We show that, under non-stress conditions, G3BP adopts a compact auto-inhibited state stabilized by electrostatic intramolecular interactions between the intrinsically disordered acidic tracts and the positively charged arginine-rich region. Upon release from polysomes, unfolded mRNAs outcompete G3BP auto-inhibitory interactions, engendering a conformational transition that facilitates clustering of G3BP through protein-RNA interactions. Subsequent physical crosslinking of G3BP clusters drives RNA molecules into networked RNA/protein condensates. We show that G3BP condensates impede RNA entanglement and recruit additional client proteins that promote SG maturation or induce a liquid-to-solid transition that may underlie disease. We propose that condensation coupled to conformational rearrangements and heterotypic multivalent interactions may be a general principle underlying RNP granule assembly.

RNA and membraneless organelles Membraneless compartments can form in cells through liquidliquid phase separation (see the Perspective by Polymenidou). But what prevents these cellular condensates from randomly fusing together? Using the RNA-binding protein (RBP) Whi3, Langdon et al. demonstrated that the secondary structure of different RNA components determines the distinct biophysical and biological properties of the two types of condensates that Whi3 forms. Several RBPs, such as FUS and TDP43, contain prion-like domains and are linked to neurodegenerative diseases. These RBPs are usually soluble in the nucleus but can form pathological aggregates in the cytoplasm. Maharana et al. showed that local RNA concentrations determine distinct phase separation behaviors in different subcellular locations. The higher RNA concentrations in the nucleus act as a buffer to prevent phase separation of RBPs; when mislocalized to the cytoplasm, lower RNA concentrations trigger aggregation. Science , this issue p. 922 , p. 918 ; see also p. 859

INTRODUCTION The formation of dynamic, membraneless compartments using intracellular phase transitions such as phase separation and gelation provides an efficient way for cells to respond to environmental changes. Recent work has identified a special class of intrinsically disordered domains enriched for polar amino acids such as glycine, glutamine, serine, or tyrosine as potential drivers of phase separation in cells. However, more traditional work has highlighted the ability of these domains to drive the formation of fibrillar aggregates. Such domains are also known as prion domains. They have first been identified in budding yeast proteins that form amyloid-like aggregates. Because these aggregates are heritable and change the activity of the prion-domain–containing protein, they are thought to be a common mechanism for phenotypic inheritance in fungi and other organisms. However, the aggregation of prion domains has also been associated with neurodegenerative diseases in mammals. Therefore, the relationship between the role of these domains as drivers of phase separation and their ability to form prion-like aggregates is unknown. RATIONALE The budding yeast translation termination factor Sup35 is an archetypal prion-domain–containing protein. Sup35 forms irreversible heritable aggregates, and these aggregates have been proposed to be either a disease or an adaptation that generates heritable phenotypic variation in populations of budding yeast. Despite having been described almost 25 years ago, the physiological functions of the Sup35 prion domain and other prion-like domains remain unclear. Uncovering these functions is a prerequisite for understanding the evolutionary pressures shaping prion-like sequences and how their physiological and pathological transitions affect cellular fitness. RESULTS Here, we show that the prion domain of Sup35 drives the reversible phase separation of the translation termination factor into biomolecular condensates. These condensates are distinct and different from fibrillar amyloid-like prion particles. Combining genetic analysis in cells with in vitro reconstitution protein biochemistry and quantitative biophysical methods, we demonstrate that Sup35 condensates form by pH-induced liquid-like phase separation as a response to sudden stress. The condensates are liquid-like initially but subsequently solidify to form protective protein gels. Cryo–electron tomography demonstrates that these gel-like condensates consist of cross-linked Sup35 molecules forming a porous meshwork. A cluster of negatively charged amino acids functions as a pH sensor and regulates condensate formation. The ability to form biomolecular condensates is shared among distantly related budding yeast and fission yeast. This suggests that condensate formation is a conserved and ancestral function of the prion domain of Sup35. In agreement with an important physiological function of the prion domain, the catalytic guanosine triphosphatase (GTPase) domain of the translation termination factor Sup35 readily forms irreversible aggregates in the absence of the prion domain. Consequently, cells lacking the prion domain exhibit impaired translational activity and a growth defect when recovering from stress. These data demonstrate that the prion domain rescues the essential GTPase domain of Sup35 from irreversible aggregation, thus ensuring that the translation termination factor remains functional during harsh environmental conditions. CONCLUSION The prion domain of Sup35 is a highly regulated molecular device that has the ability to sense and respond to physiochemical changes within cells. The N-terminal prion domain provides the interactions that drive liquid phase separation. Phase separation is regulated by the adjacent stress sensor. The synergy of these two modules enables the essential translation termination factor to rapidly form protective condensates during stress. This suggests that prion domains are protein-specific stress sensors and modifiers of protein phase transitions that allow cells to respond to specific environmental conditions.

The prevalence of morbid obesity and its co-morbidities is dramatically increasing, as is the extent of weight loss surgery. A large number of patients after various bariatric procedures need revisional intervention for various reasons. We investigated the efficacy and the safety of revisional laparoscopic Roux Y gastric bypass (LRYGB) among our patients, who were revised as a consequence of inadequate weight loss or weight regain after previous bariatric interventions. A comparative, double-centre, match pair study was performed comparing the data of 44 patients after revisional surgery with 44 patients after primary gastric bypasses, focusing on weight loss, life quality and improvement of co-morbidities. Matching criteria were age, gender, preoperative BMI and follow-up period. Previous procedures consisted of 23 gastric bandings, 13 sleeve resections, 4 LRYGB and 4 vertical banded gastroplasties. Extra weight loss (EWL) was significantly reduced after revisional gastric bypasses compared to primary intervention (EWL 66 vs. 91 %, p

PURPOSE Eye care practitioners (ECPs) acknowledge that their patients do not always follow recommendations for lens replacement, but many may not realize the possible implications for their offices. The study was conducted to investigate whether there is a relationship between contact lens compliance and the interval between full eye examinations (IEE). METHODS The study was conducted in ECP offices in the United States. Eye care practitioners and patients independently completed linked questionnaires, evaluating their contact lens wear and care. Patients were required to be current wearers of daily disposable (DD) lenses or reusable silicone hydrogel lenses with a manufacturer-recommended replacement frequency (MRRF) of 2 weeks (2WR) or 1 month (1MR). RESULTS A total of 2147 questionnaires from 141 offices were eligible. Fifty-four percent of patients were wearing 2WR, 37% 1MR, and 9% DD lenses. Wearers of 2WR lenses were significantly less compliant with replacement than wearers of both DD and 1MR lenses (34% vs. 74% and 67%, both p < 0.001); patients purchasing an annual supply were more compliant (55% vs. 45%, p < 0.001). The mean IEE was 16 months and was longer for wearers who were noncompliant with the MRRF (17.4 months vs. 14.5 months, p < 0.001). Other factors affecting IEE were household income (p = 0.030), insurance (p < 0.001), purchase source (p < 0.001), and sex (p = 0.007). CONCLUSIONS Patients who were not compliant with the MRRF had longer IEEs and were less likely to purchase an annual supply of lenses. Patients who purchased lenses from their ECP, had a higher household income, had eye examination insurance, and were female had shorter IEEs. Patients failing to replace their lenses when scheduled were also found to be less compliant with lens care procedures. Eye care practitioners should reinforce the importance of all aspects of lens wear and care with their patients, with the overall aim of reducing possible complications and retaining successful contact lens wearers in their offices.

Cells can enter into a dormant state when faced with unfavorable conditions. However, how cells enter into and recover from this state is still poorly understood. Here, we study dormancy in different eukaryotic organisms and find it to be associated with a significant decrease in the mobility of organelles and foreign tracer particles. We show that this reduced mobility is caused by an influx of protons and a marked acidification of the cytoplasm, which leads to widespread macromolecular assembly of proteins and triggers a transition of the cytoplasm to a solid-like state with increased mechanical stability. We further demonstrate that this transition is required for cellular survival under conditions of starvation. Our findings have broad implications for understanding alternative physiological states, such as quiescence and dormancy, and create a new view of the cytoplasm as an adaptable fluid that can reversibly transition into a protective solid-like state. DOI: http://dx.doi.org/10.7554/eLife.09347.001, eLife digest Most organisms live in unpredictable environments, which can often lead to nutrient shortages and other conditions that limit their ability to grow. To survive in these harsh conditions, many organisms adopt a dormant state in which their metabolism slows down to conserve vital energy. When the environmental conditions improve, the organisms can return to their normal state and continue to grow. The interior of cells is known as the cytoplasm. It is very crowded and contains many molecules and compartments called organelles that carry out a variety of vital processes. The cytoplasm has long been considered to be fluid-like in nature, but recent evidence suggests that in bacterial cells it can solidify to resemble a soft glass-type material under certain conditions. When cells become dormant they stop dividing and reorganise their cytoplasm in several ways; for example, the water content drops and many essential proteins form storage compartments. However, it was not clear how cells regulate the structure of the cytoplasm to enter into or exit from dormancy. Now, Munder et al. analyse the changes that occur in the cytoplasm when baker’s yeast cells enter a dormant state. The experiments show that when yeast cells are deprived of energy – as happens during dormancy – the cytoplasm becomes more acidic than normal. This limits the ability of molecules and organelles to move around the cytoplasm. Similar results were also seen in other types of fungi and an amoeba. Munder et al. found that this increase in acidity during dormancy causes many proteins to interact with each other and form large clumps or filament structures that result in the cytoplasm becoming stiffer. A separate study by Joyner et al. found that when yeast cells are starved of sugar, two large molecules are less able to move around the cell interior. Together, the findings of the studies suggest that the interior of cells can undergo a transition from a fluid-like to a more solid-like state to protect the cells from damage when energy is in short supply. The next challenge is to understand the molecular mechanisms that cause the physical properties of the cytoplasm to change under different conditions. DOI: http://dx.doi.org/10.7554/eLife.09347.002

The deposition of misfolded proteins in cytoplasmic protein bodies requires the concerted action of stress-inducible protein-sorting factors and molecular chaperones. Protein sequestration during acute stress is a cellular strategy that adjusts the flux of misfolded proteins to the capacities of the protein quality control system., Acute stress causes a rapid redistribution of protein quality control components and aggregation-prone proteins to diverse subcellular compartments. How these remarkable changes come about is not well understood. Using a phenotypic reporter for a synthetic yeast prion, we identified two protein-sorting factors of the Hook family, termed Btn2 and Cur1, as key regulators of spatial protein quality control in Saccharomyces cerevisiae. Btn2 and Cur1 are undetectable under normal growth conditions but accumulate in stressed cells due to increased gene expression and reduced proteasomal turnover. Newly synthesized Btn2 can associate with the small heat shock protein Hsp42 to promote the sorting of misfolded proteins to a peripheral protein deposition site. Alternatively, Btn2 can bind to the chaperone Sis1 to facilitate the targeting of misfolded proteins to a juxtanuclear compartment. Protein redistribution by Btn2 is accompanied by a gradual depletion of Sis1 from the cytosol, which is mediated by the sorting factor Cur1. On the basis of these findings, we propose a dynamic model that explains the subcellular distribution of misfolded proteins as a function of the cytosolic concentrations of molecular chaperones and protein-sorting factors. Our model suggests that protein aggregation is not a haphazard process but rather an orchestrated cellular response that adjusts the flux of misfolded proteins to the capacities of the protein quality control system.

The laparoscopic Roux Y gastric bypass (LRYGB) is one of the most often performed bariatric surgical intervention. Intraoperative gastroscopy (IOG) seems to be reliable to decrease the leakage rate of gastrojejunal anastomosis (GJA) and of gastric pouch (GP). Our aim was to test the efficacy and the safety of this method. Two hundred fifty-two LRYGB operations were performed in our institution between 1 January 2008 and 1 January 2010. IOG is routinely made to test the integrity of GJA and of GP. Patients' dates were retrospectively analysed. The intragastric pressure developed during gastroscopy in humans was measured and compared with pressure values led to destruction (positive air test) of the GJA and/or GP in animal models (hybrid pigs). Stomach and bowel wall samples from the test animals without pressure strain, with pressure strain developed at gastroscopy in humans and with pressure strains led to destruction of GJA and/or GP were histologically examined. IOG resulted in six of our cases (2.3%) positive air test. There was no anastomosis insufficiency in postoperative period. Mean pressure during IOG was 32 mmHg, mean time of examination was 3.8 min and mean maximal pressure was 43 mmHg in humans. The mean pressure leading to positive air test in pigs was 150 mmHg. We could not detect any microscopical difference between stomach and jejunum samples without pressure strain and after pressure strain developed in humans during the gastroscopy. We conclude that intraoperative gastroscopy is an effective and safe method to test the integrity of GJA and GP in LRYGB surgery.

Lipid homeostasis is essential for proper function of cells and organisms. To unravel new regulators of this system, we developed a screening procedure, combining RNA interference in HeLa cells and TLC, which enabled us to monitor modifications of lipid composition resulting from short, interfering RNA knock-downs. We applied this technique to the analysis of 600 human kinases. Despite the occurrence of off-target effects, we identified JNK2 as a new player in triglyceride (TG) homeostasis and lipid droplet metabolism and, more specifically, in the regulation of lipolysis. Similar control of the level of TGs and lipid droplets was observed for its Schizosaccharomyces pombe homolog, Sty1, suggesting an evolutionary conserved function of mitogen-activated protein kinases in the regulation of lipid storage in eukaryotic cells.

One of the key questions in biology is how the metabolism of a cell responds to changes in the environment. In budding yeast, starvation causes a drop in intracellular pH, but the functional role of this pH change is not well understood. Here, we show that the enzyme glutamine synthetase (Gln1) forms filaments at low pH and that filament formation leads to enzymatic inactivation. Filament formation by Gln1 is a highly cooperative process, strongly dependent on macromolecular crowding, and involves back-to-back stacking of cylindrical homo-decamers into filaments that associate laterally to form higher order fibrils. Other metabolic enzymes also assemble into filaments at low pH. Hence, we propose that filament formation is a general mechanism to inactivate and store key metabolic enzymes during a state of advanced cellular starvation. These findings have broad implications for understanding the interplay between nutritional stress, the metabolism and the physical organization of a cell. DOI: http://dx.doi.org/10.7554/eLife.02409.001, eLife digest Life is based on a series of chemical reactions that control how cells live, grow, and divide. Various metabolic enzymes allow cells to control the rate at which these reactions occur. Recently, researchers have noticed that metabolic enzymes can form filaments in cells, usually when the cells are deprived of energy or nutrients. Petrovska, Nüske et al. now reveal more about how and why an enzyme called glutamine synthetase (Gln1) forms filaments in yeast cells. Gln1 has a cylindrical shape. This shape means that stacking the enzymes end-to-end under the right conditions is enough to make them bond into a long filament. In addition, a zip-like mechanism enables neighboring filaments to fuse to create thicker fibres. These filaments are more likely to form if there are high concentrations of large background molecules around—a condition known as macromolecular crowding. Petrovska, Nüske et al. also found evidence that the filaments are part of a strategy to help cells survive starvation. Enzymes were more likely to construct filaments when the cell division cycle had stopped, which commonly occurs due to a lack of nutrients. In addition, Gln1 filaments only form if the cytoplasm of the cell becomes acidic—which is a response to the cell starving. This has been seen for other metabolic enzymes as well, suggesting that acidification is a signal to reprogram the metabolism of a cell. The Gln1 enzymes in a filament are inactivated, but become active again after the filament breaks up. In addition, preventing Gln1 filament formation makes it harder for cells to recover from a period of starvation. Petrovska, Nüske et al. therefore suggest that the filaments act as a storage depot for the enzymes during starvation. Further experiments are now needed to uncover exactly how these manage to help the starving cell to survive and recover. DOI: http://dx.doi.org/10.7554/eLife.02409.002

This chapter presents Body-centred activities in Mbembe (Benue-Congo, Central Jukunoid) to include physical processes as well as cognition, perception and emotion. These can be referred to by different classes of verbs. Certain verbs are specific to one body-centred activity, while others can describe different activities or be used in other contexts. Depending on the class of the verb as well as on the meaning expressed in the respective context the arguments are realised in different ways. For example, verbs which can be used in contexts other than body-centred activities often require an explicit reference to the body of the referent performing or being affected by the activity. Further in-depth studies are needed to fully understand expressions of body centred activities in Mbembe, especially in the cases where verbs of different semantic classes combine to describe new meanings. Keywords: body-centred activities; cognition; emotion; mbembe; perception; verb

To collect questionnaire data from patients in a large clinical population that would allow for an estimate of the prevalence of self-reported symptoms of dry eyes.A 13-point questionnaire (The Canada Dry Eye Epidemiology Study, CANDEES) was mailed to all optometric practices in Canada in October 1994, with the request that it be completed by 30 successive nonselected patients.Four hundred fifty sets of questionnaires (total, 13,517) were analyzed (a 15.7% return rate) from patients aged from10 years to80 years; 55% were in the 21- to 50-year age group; 60.7% were female; and 24.3% were contact lens wearers. A total of 28.7% reported dry eye symptoms, of whom 24.2% reported concurrent dry mouth, 24.5% had worse symptoms in the morning, 30.3% reported concurrent lid problems, and 35.7% reported a history of allergies. Of the 3716 patients reporting symptoms, 62 (1.6%) were in the "severe" category and 290 (7.8%) were in the "constant but moderate" category. Contact lens wear, concurrent allergies, dry mouth, lid problems, or use of medications increased the chance of a patient reporting dry eye symptoms.The prevalence of patients reporting any level of symptoms of dry eyes was approximately 1 in 4; severe symptoms were reported by 1 in 225 patients.

This study assessed the repeatability of the Nikon NRK-8000, the Nidek AR-1000, and subjective refraction. In addition, the accuracy of the Nikon and the Nidek were evaluated in comparison to subjective refraction. Measurements were taken with all 3 techniques on 2 separate occasions with a test-retest separation of at least 24 h. The right eyes of 30 normal subjects were used. Repeatability and accuracy statistics and plots were generated using matrix representations of dioptric power. Subjective refraction was the most repeatable method, with the coefficient of repeatability (COR) found to be 0.611, 0.224, and 0.490 in the vertical, torsional, and horizontal meridians. The autorefractors' COR was found to range from 0.712 to 0.826 for the vertical and horizontal meridians, whereas the torsional meridian ranged from 0.224 to 0.319.

Purpose. To evaluate subjective ocular comfort across the day with three silicone hydrogel daily disposables (SHDDs) in a group of adapted lens wearers. Methods. Masked subjects (asymptomatic or symptomatic of end-of-day (EOD) dryness with habitual lenses) wore three SHDDs: DAILIES TOTAL1 (DT1), Clariti 1day (C1D), or 1-DAYACUVUE TRUEYE (AVTE), each for 3 days. On day 2, wearing time (WT) and comfort ratings after insertion, at 4, 8, and 12 hours, and at EOD were recorded. Because not all subjects wore lenses for 12 hours, comfort was analyzed across the day (up to 8 hours, 8 to 12 hours), and a new variable (‘‘cumulative comfort’’ [CC]) was calculated for EOD. Results. One hundred four subjects completed the study (51 asymptomatic, 53 symptomatic). The two groups had different WTs (mean WT, 14.0 and 12.7 hours, respectively; p G 0.001). Ocular comfort was rated higher in the asymptomatic group throughout the day (p G 0.001). One hundred four subjects wore all three SHDDs for at least 8 hours, whereas 74 (45 asymptomatic, 29 symptomatic) subjects wore them for 12 hours or longer. Comfort ratings were higher with DT1 (least square means [LSM] = 91.0) than with C1D (LSM = 86.5; p G 0.001) and AVTE (LSM = 87.7; p = 0.011) for the first 8 hours and lower with C1D compared with DT1 (p = 0.012) from 8 to 12 hours. Mean EOD (T SD) comfort with the C1D lens was 72T 21, lower than both DT1 (mean, 79T 17; p = 0.001) and AVTE (mean, 78T 21; p = 0.010). Mean CC was higher in the asymptomatic group (mean, 1261 T 59) compared with that in the symptomatic group (mean, 1009 T 58; p G 0.001) and higher for DT1 (mean, 1184 T 258) than C1D (mean, 1094 T 318; p = 0.002) and AVTE (mean, 1122 T 297; p = 0.046). Conclusions. All three SHDDs had average WTs of 12 hours or longer for 1 day. Comfort during the first 12 hours was highest with DT1 (similar to AVTE between 8 and 12 hours) and lowest with C1D. End-of-day comfort was lowest with C1D, and CC was highest for DT1. Cumulative comfort may be a valuable new metric to assess ocular comfort during the day. (Optom Vis Sci 2013;90:945Y953)

Purpose To investigate compliance with daily disposable contact lens (DDCL) wear and investigate re-use of lenses according to country and DDCL material worn. Methods Optometrists invited eligible DDCL patients from their practices to participate in a survey on DDCL wear in Australia, Norway, the United Kingdom (UK) and the United States (US). Eligible participants completed an online or paper version of the survey. Results 805 participants completed the survey (96% online): Australia 13%, Norway 32%, UK 17%, US 38%. The median age was 38 years; 66% were female. Silicone hydrogel (SiHy) DDCLs were worn by 14%. Overall, 9% were non-compliant with DDCL replacement; Australia 18%, US 12%, UK 7% and Norway 4%. There were no differences with respect to sex, years of contact lens wear experience or DDCL material (SiHy versus hydrogels). The primary reason for re-use was “to save money” (60%). Re-use of DDCLs resulted in inferior comfort at insertion and prior to lens removal ( p = 0.001). 75% reported occasional napping and 28% reported sleeping overnight for at least one night in the preceding month, while wearing their DDCLs. Conclusion Non-compliance with replacement of DDCLs occurred in all countries investigated; the rate was highest in Australia and lowest in Norway. Re-use of DDCLs was associated with reduced comfort. DDCL wearers often reported wearing lenses overnight. It is important for optometrists to counsel their patients on the importance of appropriate lens wear and replacement for DDCLs.

PURPOSE To determine distribution of central corneal swelling (CCS) across subjects after 8 hr of sleep in eyes wearing silicone hydrogel lenses with various oxygen transmissibility (Dk/t) values and in eyes without lenses. METHODS Twenty-nine neophytes wore lotrafilcon A (Dk, 140), balafilcon A (Dk, 91), galyfilcon A (Dk, 60), and senofilcon A (Dk, 103) lenses in powers -3.00, -10.00, and +6.00 diopters on separate nights, in random order, and on 1 eye only. The contralateral eye (no lens) served as the control. Central corneal thickness was measured using a digital optical pachometer before lens insertion and immediately after lens removal on waking. RESULTS The average difference between the mean (7%) and the median (6.8%) CCS of all lenses was only 0.2%, suggesting a normal distribution. There was no correlation between the mean and the range of the CCS (r=0.058, P=0.766). Normal CCS distributions were also found with each lens-wearing eye and the control eye (P>0.20 for all). There was a significant correlation between lens-wearing eye and control eye (r=0.895, P

Purpose. Silicone hydrogel (SH) lenses are usually replaced after 2 weeks (2W) or 1 month (1M); however, many patients do not comply with the manufacturers’ recommended replacement frequency (MRRF). The purpose of this analysis was to investigate the effect of compliance with MRRF on comfort and vision in SH wearers. Methods. As a part of a larger study investigating compliance with MRRF, patients were asked to rate their subjective comfort and vision from 0 (very poor) to 10 (excellent) in the morning, at the end of the day (EVE), when lenses were new, and needed replacing (NR). Results. One thousand three hundred forty-four patients wore 2W replacement modality (2WR) (n 717) or 1M replacement modality (1MR) (n 617) SH lenses. Comfort and vision in the morning and when lenses are new were significantly higher than for EVE and NR (p 0.001). Twenty-nine percent (95% confidence interval 25.3–32.4) of 1MR and 59% (95% confidence interval 55.5–62.7) of 2WR wearers were non-compliant with the MRRF. Compliance had a significant effect on EVE (p 0.002, p 0.008) and NR (p 0.001, p 0.001) comfort and vision. After accounting for compliance, EVE and NR comfort and EVE vision were higher for 1MR than 2WR (p 0.015, p 0.044, p 0.019). Conclusions. Compliant patients had better EVE and NR comfort and vision than non-compliant patients, regardless of replacement modality. Optimal subjective performance with SH lenses seems to be facilitated by replacing lenses as recommended. (Optom Vis Sci 2010;87:1–●●●)

To assess eye care practitioners (ECPs) recommendations for replacement frequency (RF) of silicone hydrogel (SH) and daily disposable (DD) lenses in Canada and the U.S. and to compare noncompliance (NC) with manufacturer recommended RF by the ECP and patient, and the reasons given for NC.Invitations to participate were sent by e-mail to ECPs in Canada and the U.S. Twenty patient surveys were sent to 420 ECPs, and 2232 eligible surveys were received from 216 ECPs (26% Canada, 74% U.S.). Questions related to patient demographics, lens type, wearing patterns, ECP instructions for RF, and actual patient RF. ECPs provided lens information and their recommendation for RF after the surveys were completed and sealed in envelopes. Responses were anonymous.DD accounted for 18% (Canada) vs. 16% (U.S.) of wearers (p0.05); 35% (Canada) vs. 45% (U.S.) wore 2-week replacement SH (2WR; p = 0.011); and 47% (Canada) vs. 39% (U.S.) wore 1-month replacement SH (1MR) lenses (p = 0.025). Thirty-four percent (Canada) vs. 18% (U.S.) of ECPs recommended longer RFs than the manufacturer recommended RF for 2WR lens wearers (p0.001); 6% (Canada) vs. 4% (U.S.) for DD wearers; and 2% (Canada) vs. 1% (U.S.) for 1MR lens wearers. NC rates for actual RFs reported by patients were not different between countries (p0.05) and were lowest for DD (13% Canada, 12% U.S.), followed by 1MR (33% Canada, 28% U.S.). The highest NC rates were with 2WR (50% Canada, 52% U.S.). The most frequent reason for NC with 2WR and 1MR was "forgetting which day to replace lenses" (54% Canada, 53% U.S.) and in DD wearers "to save money" (56% Canada, 29% U.S., p0.001).1MR lenses are more frequently prescribed in Canada. ECPs in Canada were NC with 2WR lenses more frequently than U.S. ECPs, but patient NC rates were the same in both countries for all lens types. ECP and patient NC rates were highest for 2WR lens wearers.

Purpose: A four-month extended wear clinical trial was conducted to compare the ocular effects of a high Dk Balafilcon A silicone hydrogel lens and a low Dk HEMA 38.6 per cent H20 soft lens. Method: Twenty-four subjects who were adapted to daily wear of soft lenses wore a high Dk lens in one eye and a low Dk HEMA lens in the other eye for four months on an extended wear basis after one week of daily wear. Thirteen progress evaluations were conducted using standard clinical procedures. Results: Eighteen subjects (75 per cent) completed the study. The high Dk lens induced significantly less bulbar and limbal injection and corneal vascularisation than the low Dk HEMA lens (p < 0.05). Epithelial microcysts were observed only in the eyes wearing the low Dk lens. A significant increase in myopia was found in the eyes wearing the low Dk HEMA lens (mean = 0.50 D, p < 0.01) compared to the insignificant myopic increase of 0.06 D in the eyes wearing the high Dk lens. Three subjects developed small infiltrates in the high Dk lens wearing eyes and significantly more post-lens debris was observed under the high Dk lens. Six subjects developed papillary conjunctivitis in the eye wearing silicone hydrogel lenses but only two of those were discontinued from the study. Conclusion: No hypoxia-related effects were observed with extended wear of the high Dk Balafilcon A silicone hydrogel lens.

Purpose. The purpose of this study was to determine the impact of extended wear of high and low oxygen permeability (Dk) lenses on the development or resolution of limbal hyperemia and corneal neovascularization in a randomized prospective clinical trial. Methods. Adapted daily-wear contact lens wearers were randomly assigned to one of two groups. The low-Dk group (N = 23) wore etafilcon A lenses (Dk/t = 40) for up to 7 days and 6 nights, and the high-Dk group (N = 39) wore lotrafilcon A lenses (Dk/t = 175) for up to 30 days and nights. Limbal hyperemia and neovascularization were assessed and graded during 9 months of extended wear. A stratified analysis to determine how change in ocular redness depended on initial presentation was also conducted. Results. On a 0 to 100 scale, extended wear of low-Dk lenses resulted in a 16-point increase in limbal hyperemia (p < 0.0001); no significant change occurred with the high-Dk lenses. The difference was greatest for low-Dk participants who initially presented with lower levels of hyperemia (N = 18). There was a slight resolution of redness in participants who initially presented with higher levels of hyperemia (N = 10) after wearing the high-Dk lenses. Neovascularization increased by an average of 0.5 on a scale of 0 to 4 in low-Dk lens wearers (p < 0.0001) but remained unchanged in the high-Dk lens wearers. The increase was most marked for the low-Dk group with lower levels of neovascularization at baseline. Conclusions. Subjects who wore lenses with higher oxygen permeability for 9 months of extended wear exhibited a lower vascular response of the ocular surface compared with subjects who wore lenses of lower oxygen permeability for the same period. This result was found both for the surface limbal vessels and in the deeper neovascular stromal vessels. Moderate neovascularization also developed after 3 months of extended wear of low-Dk hydrogels, whereas high-Dk lenses caused no neovascularization.

A small but significant increase in myopia after extended wear of low oxygen permeability (Dk) hydrogel lenses has been previously reported; however, the specific impact of hypoxia on refractive status and corneal curvature with extended wear are not well documented. The purpose of this study was to compare the refractive changes induced over a period of 9 months' extended wear with high-Dk fluorosiloxane hydrogel lenses and low-Dk hydrogel lenses.Adapted daily wear contact lens wearers were randomly assigned to one of two groups. The low-Dk group wore etafilcon A (Dk = 28) for up to 7 days and 6 nights and the high-Dk group wore lotrafilcon A (Dk = 140) for up to 30 days and nights. Refractive error and corneal curvature were measured at 3-month intervals over 9 months of extended wear.The etafilcon A group demonstrated an average increase in myopia of 0.30 D over the 9-month period; however, no change in spherical myopic correction was measured in the lotrafilcon A group. The cylindrical component did not change in either group. A stratified analysis revealed a greater increase in myopia for low myopes than moderate myopes in the etafilcon A group but no difference in the lotrafilcon A group. Keratometric analysis revealed no change in the etafilcon A group and a small degree of central corneal flattening in both major meridians of 0.35 D in the lotrafilcon A group.Nine months of extended wear of low-Dk lenses is associated with a small degree of myopic progression in adult myopes that appears to be reversible. Wearing fluorosiloxane-hydrogel lenses of high-Dk had no impact on refractive error and may be associated with a small degree of central corneal flattening.

This study assessed the repeatability of the Nikon NRK-8000, the Nidek KM-800 and the Bausch and Lomb keratometer. In addition, the comparability of the Nikon NRK-8000 and the Nidek KM-800 are evaluated relative to the Bausch and Lomb keratometer. Measurements were taken with all three techniques on two separate occasions with a test-retest separation of at least 24 hr. The right eyes of 30 normal subjects were used. Repeatability and comparability statistics and plots were generated using matrix representations of dioptric power. The Nidek KM-800 coefficient of repeatability values were found to be 0.345, 0.187, and 0.321 in the vertical, torsional, and horizontal meridians while the Nikon NRK-8000 values were 0.346, 0.232, and 0.276 respectively.