High-throughput methylome analysis reveals differential methylation for early and late onset preeclampsia for mothers and their children.

Preeclampsia is a hypertensive disorder of pregnancy that affects ∼2%-5% of all pregnancies, contributes to 4 of the top 10 causes of pregnancy-related deaths, and remains a long-term risk factor for cardiometabolic diseases. Yet, little is still known about the molecular mechanisms that lead to this disease. There is evidence that some cases have a genetic cause. However, it is well appreciated that harmful factors in the environment, such as poor nutrition, stress, and toxins, may lead to epigenetics changes that can contribute to this disease. DNA methylation is one of the epigenetic modifications known to be fairly stable and impact gene expression. Using DNA from buccal swabs, we analyzed global DNA methylation among three groups of individuals: mothers who experienced 1) early-stage preeclampsia (<32 wk), 2) late-stage preeclampsia (>37 wk), or 3) no complications during their pregnancies, as well as the children from these three groups. We found significant differentially methylated regions (DMRs) between mothers who experienced preeclampsia compared with those with no complications adjacent or within genes that are important for placentation, embryonic development, cell adhesion, and inflammation (e.g., the cadherin pathway). A significant portion of DMR genes showed expression in tissues relevant to preeclampsia (i.e., the brain, heart, kidney, uterus, ovaries, and placenta). As this study was performed on DNA extracted from cheek swabs, this opens the way to future studies in different tissues, aimed at identifying possible biomarkers of risk and early detection, developing targeted interventions, and reducing the progression of this life-threatening disease.NEW & NOTEWORTHY Preeclampsia is a life-threatening hypertensive disorder, affecting 2%-5% of pregnancies, that remains poorly understood. This study analyzed DNA methylation from buccal swabs from mothers who experienced early and late-stage preeclampsia and those with uncomplicated pregnancies, along with their children. Differentially methylated regions were found near and within genes crucial for placental function, embryonic development, and inflammation. Many of these genes are expressed in preeclampsia-related tissues, offering hope for future biomarker development for this condition.

Grandmaternal allergen sensitization reprograms epigenetic and airway responses to allergen in second-generation offspring.

Asthma susceptibility is influenced by environmental, genetic, and epigenetic factors. DNA methylation is one form of epigenetic modification that regulates gene expression and is both inherited and modified by environmental exposures throughout life. Prenatal development is a particularly vulnerable time period during which exposure to maternal asthma increases asthma risk in offspring. How maternal asthma affects DNA methylation in offspring and what the consequences of differential methylation are in subsequent generations are not fully known. In this study, we tested the effects of grandmaternal house dust mite (HDM) allergen sensitization during pregnancy on airway physiology and inflammation in HDM-sensitized and challenged second-generation mice. We also tested the effects of grandmaternal HDM sensitization on tissue-specific DNA methylation in allergen-naïve and -sensitized second-generation mice. Descendants of both allergen- and vehicle-exposed grandmaternal founders exhibited airway hyperreactivity after HDM sensitization. However, grandmaternal allergen sensitization significantly potentiated airway hyperreactivity and altered the epigenomic trajectory in second-generation offspring after HDM sensitization compared with HDM-sensitized offspring from vehicle-exposed founders. As a result, biological processes and signaling pathways associated with epigenetic modifications were distinct between lineages. A targeted analysis of pathway-associated gene expression found that Smad3 was significantly dysregulated as a result of grandmaternal allergen sensitization. These data show that grandmaternal allergen exposure during pregnancy establishes a unique epigenetic trajectory that reprograms allergen responses in second-generation offspring and may contribute to asthma risk.NEW & NOTEWORTHY Asthma susceptibility is influenced by environmental, genetic, and epigenetic factors. This study shows that maternal allergen exposure during pregnancy promotes unique epigenetic trajectories in second-generation offspring at baseline and in response to allergen sensitization, which is associated with the potentiation of airway hyperreactivity. These effects are one mechanism by which maternal asthma may influence the inheritance of asthma risk.

Co-option of the lineage-specific LAVA retrotransposon in the gibbon genome.

Co-option of transposable elements (TEs) to become part of existing or new enhancers is an important mechanism for evolution of gene regulation. However, contributions of lineage-specific TE insertions to recent regulatory adaptations remain poorly understood. Gibbons present a suitable model to study these contributions as they have evolved a lineage-specific TE called LAVA (LINE-AluSz-VNTR-AluLIKE), which is still active in the gibbon genome. The LAVA retrotransposon is thought to have played a role in the emergence of the highly rearranged structure of the gibbon genome by disrupting transcription of cell cycle genes. In this study, we investigated whether LAVA may have also contributed to the evolution of gene regulation by adopting enhancer function. We characterized fixed and polymorphic LAVA insertions across multiple gibbons and found 96 LAVA elements overlapping enhancer chromatin states. Moreover, LAVA was enriched in multiple transcription factor binding motifs, was bound by an important transcription factor (PU.1), and was associated with higher levels of gene expression in cis We found gibbon-specific signatures of purifying/positive selection at 27 LAVA insertions. Two of these insertions were fixed in the gibbon lineage and overlapped with enhancer chromatin states, representing putative co-opted LAVA enhancers. These putative enhancers were located within genes encoding SETD2 and RAD9A, two proteins that facilitate accurate repair of DNA double-strand breaks and prevent chromosomal rearrangement mutations. Co-option of LAVA in these genes may have influenced regulation of processes that preserve genome integrity. Our findings highlight the importance of considering lineage-specific TEs in studying evolution of gene regulatory elements.

Assessment of Metabolic Flexibility by Substrate Oxidation Responses and Blood Lactate in Women Expressing Varying Levels of Aerobic Fitness and Body Fat.

ABSTRACT: Waldman, HS, Bryant, AR, Knight, SN, Killen, LG, Davis, BA, Robinson, MA, and O'Neal, EK. Assessment of metabolic flexibility by substrate oxidation responses and blood lactate in women expressing varying levels of aerobic fitness and body fat. J Strength Cond Res 37(3): 581-588, 2023-Collection of substrate oxidation responses during exercise is proposed as a noninvasive means for assessing metabolic flexibility in male subjects. However, because of hormonal and metabolic differences between sexes, this method may not be applicable to female subjects. This study assessed metabolic flexibility through indirect calorimetry across female subjects with different maximal oxidative capacities. Thirty-eight (18-45 years) eumenorrheic female subjects were stratified ( p < 0.05) based on V̇ o2 peak (mL·kg -1 ·min -1 ) into (1) endurance-trained (ET, n = 12, 42.6 ± 5.3), (2) recreationally active (RA, n = 13, 32.3 ± 1.6), or (3) overweight female subjects (OW, n = 13, 21.0 ± 4.0). Subjects completed the same 5-stage graded exercise test with intensities of 30, 45, 60, 75, and 90 W. Lactate [La - ], carbohydrate (CHOox), and fat (FATox) oxidation rates were assessed during the last min of each 5-minute stage. Subjects then cycled to exhaustion to determine V̇ o2 peak. Endurance-trained and RA female subjects expressed significantly ( p ≤ 0.05) higher absolute rates and rates scaled to fat-free mass of CHOox and FATox compared with OW female subjects during multiple stages. [La - ] failed to consistently differentiate the 3 groups with higher [La - ] for OW only found during stage 4; however, RER differed by 0.09 units or more at each stage for OW vs. ET. It seems that RER was more sensitive to cohort characteristics than [La - ] contrasting recent findings in male cohorts. In conclusion, indirect calorimetry is a practical and noninvasive method for assessing metabolic flexibility in eumenorrheic female subjects of varying aerobic fitness levels.

Axial elongation associated with biomechanical factors during near work.

PURPOSE: To investigate the changes occurring in the axial length, choroidal thickness, and anterior biometrics of the eye during a 10-minute near task performed in downward gaze. METHODS: Twenty young adult subjects (10 emmetropes and 10 myopes) participated in this study. To measure ocular biometrics in downward gaze, an optical biometer was inclined on a custom-built height- and tilt-adjustable table. Baseline measures were collected after each subject performed a distance primary gaze control task for 10 minutes to provide washout period for previous visual tasks before each of three different accommodation/gaze conditions. These other three conditions included a near task (2.5 diopters [D]) in primary gaze and a near (2.5 D) and a far (0 D) accommodative task in downward gaze (25 degrees), all for 10 minutes' duration. Immediately after and then 5 and 10 minutes from the commencement of each trial, measurements of ocular biometrics (e.g., anterior biometrics, axial length, choroidal thickness, and retinal thickness) were obtained. RESULTS: Axial length increased with accommodation and was significantly greater for downward gaze with accommodation (mean ± SD change, 23 ± 13 µm at 10 minutes) compared with primary gaze with accommodation (8 ± 15 µm at 10 minutes) (p < 0.05). A small amount of choroidal thinning was also found during accommodation that was statistically significant in downward gaze (13 ± 14 µm at 10 minutes; p < 0.05). Accommodation in downward gaze also caused greater changes in anterior chamber depth and lens thickness compared with accommodation in primary gaze. CONCLUSIONS: Axial length, choroidal thickness, and anterior eye biometrics change significantly during accommodation in downward gaze as a function of time. These changes seem to be caused by the combined influence of biomechanical factors (i.e., extraocular muscle forces, ciliary muscle contraction) associated with near tasks in downward gaze.

24 h Hydration profile of collegiate soccer players training once versus twice per day in the heat.

This study examined 24-h hydration parameters among collegiate, male soccer players (n = 17) during twice (X2) and once (X1) per day practice schedules in the heat. Urine specific gravity (USG) and body mass were measured before morning practices, afternoon practice (X2)/team meeting (X1), and the next morning practices. Fluid intake, sweat losses, and urinary losses were assessed during each 24-h window. Pre-practice body mass or USG did not differ among the timepoints. Sweat losses differed among all practices (p < 0.05) and averaged approximately 2.181±0.693 (X2AM) 1.710±0.474 (X2PM), and 3.361±0.956 L (X1AM), but players averaged replacing >50% of sweat losses with fluid intake every practice. Fluid intake during and between practices from practice 1 to the afternoon practice for X2 resulted in a positive fluid balance for X2 (+0.446±0.916 L). However, higher sweat loss during the initial morning practice and lower relative fluid intake prior to the afternoon team meeting the following morning resulted in a negative fluid balance (-0.304±0.675 L; p < 0.05: Cohen's d = 0.94) over the same time period for X1. By the start of the next morning practice sessions, both X1 (+0.664±1.051 L) and X2 (+0.446±0.916 L) were in positive fluid balance, respectively. Ample fluid consumption opportunities, scaled down practice intensities during X2, and potentially intentional greater relative fluid intake during X2 training resulted in no difference in fluid shift versus an X1 schedule before the start of practices. The majority of players maintained fluid balance drinking ad libitum regardless of practice schedule.

Runners greatly underestimate sweat losses before and after a 1-hr summer run.

The purpose of this study was to determine how accurately runners estimate their sweat losses. Male (n = 19) and female (n = 20) runners (41 ± 10 yr, VO2max 57 ± 9 ml · kg(-1) · min(-1) from the southeastern U.S. completed an ~1-hr run during late summer on a challenging outdoor road course (wet bulb globe temperature 24.1 ± 1.5 °C). Runs began at ~6:45 a.m. or p.m. Before and after running, participants filled race-aid-station paper cups with a volume of fluid they felt would be equivalent to their sweat losses. Total sweat losses and losses by percent body weight differed (p < .01) between men (1,797 ± 449 ml, 2.3% ± 0.6%) and women (1,155 ± 258 ml, 1.9% ± 0.4%). Postrun estimates (738 ± 470 ml) were lower (p < .001) than sweat losses (1,468 ± 484 ml), equaling underestimations of 50% ± 23%, with no differences in estimation accuracy by percentage between genders. Runners who reported measuring changes in pre- and postrun weight to assess sweat losses within the previous month (n = 9, -54% ± 18%) were no more accurate (p = .55) than runners who had not (n = 30, -48% ± 24%). These results suggest that inadequate fluid intake during runs or between runs may stem from underestimations of sweat losses and that runners who do assess sweat-loss changes may be making sweat-loss calculation errors or do not accurately translate changes in body weight to physical volumes of water.

DNA Methylation Analysis of Turner Syndrome BAV.

Turner Syndrome (TS) is a rare cytogenetic disorder caused by the complete loss or structural variation of the second sex chromosome. The most common cause of early mortality in TS results from a high incidence of left-sided congenital heart defects, including bicuspid aortic valve (BAV), which occurs in about 30% of individuals with TS. BAV is also the most common congenital heart defect in the general population with a prevalence of 0.5-2%, with males being three-times more likely to have a BAV than females. TS is associated with genome-wide hypomethylation when compared to karyotypically normal males and females. Alterations in DNA methylation in primary aortic tissue are associated with BAV in euploid individuals. Here we show significant differences in DNA methylation patterns associated with BAV in TS found in peripheral blood by comparing TS BAV (n = 12), TS TAV (n = 13), and non-syndromic BAV (n = 6). When comparing TS with BAV to TS with no heart defects we identified a differentially methylated region encompassing the BAV-associated gene MYRF, and enrichment for binding sites of two known transcription factor contributors to BAV. When comparing TS with BAV to euploid women with BAV, we found significant overlapping enrichment for ChIP-seq transcription factor targets including genes in the NOTCH1 pathway, known for involvement in the etiology of non-syndromic BAV, and other genes that are essential regulators of heart valve development. Overall, these findings suggest that altered DNA methylation affecting key aortic valve development genes contributes to the greatly increased risk for BAV in TS.

The influence of downward gaze and accommodation on ocular aberrations over time.

A Shack-Hartmann wavefront sensor was modified to allow measurement of ocular aberrations in downward gaze with binocular fixation. Subjects first performed a control task prior to four different trials involving a distance task or a near task (accommodative demand of 2.5 D) performed in primary and downward gaze (25°). Immediately after beginning and then again 5 and 10 min after the commencement of each trial, ocular aberrations were measured. To observe the recovery in ocular aberrations following each test condition, subjects again viewed a distance target in primary gaze and aberration measurements were taken at 0, 5, and 10 min. During the distance task, small but significant changes in refractive power and higher order aberrations were observed in downward gaze compared to primary gaze. The changes in ocular aberrations that occurred in downward gaze recovered almost immediately after shifting gaze from downward back to primary gaze. During the accommodation tasks, there was a significant influence of gaze for changes in primary spherical aberration C(4, 0) [p=0.004] and secondary spherical aberration C(6, 0) [p=0.02]. There was also a significant gaze by time interaction (p=0.04) for changes in C(6, 0). These findings show that ocular aberrations change from primary to downward gaze, particularly during accommodation.

Gene-Specific DNA Methylation Linked to Postoperative Cognitive Dysfunction in Apolipoprotein E3 and E4 Mice.

BACKGROUND: Geriatric surgical patients are at higher risk of developing postoperative neurocognitive disorders (NCD) than younger patients. The specific mechanisms underlying postoperative NCD remain unknown, but they have been linked to genetic risk factors, such as the presence of APOE4, compared to APOE3, and epigenetic modifications caused by exposure to anesthesia and surgery. OBJECTIVE: To test the hypothesis that compared to E3 mice, E4 mice exhibit a more pronounced postoperative cognitive impairment associated with differential DNA methylation in brain regions linked to learning and memory. METHODS: 16-month-old humanized apolipoprotein-E targeted replacement mice bearing E3 or E4 were subjected to surgery (laparotomy) under general isoflurane anesthesia or sham. Postoperative behavioral testing and genome-wide DNA methylation were performed. RESULTS: Exposure to surgery and anesthesia impaired cognition in aged E3, but not E4 mice, likely due to the already lower cognitive performance of E4 prior to surgery. Cognitive impairment in E3 mice was associated with hypermethylation of specific genes, including genes in the Ephrin pathway implicated in synaptic plasticity and learning in adults and has been linked to Alzheimer's disease. Other genes, such as the Scratch Family Transcriptional Repressor 2, were altered after surgery and anesthesia in both the E3 and E4 mice. CONCLUSION: Our findings suggest that the neurocognitive and behavioral effects of surgery and anesthesia depend on baseline neurocognitive status and are associated with APOE isoform-dependent epigenetic modifications of specific genes and pathways involved in memory and learning.

Integrated analysis of behavioral, epigenetic, and gut microbiome analyses in AppNL-G-F, AppNL-F, and wild type mice.

Epigenetic mechanisms occurring in the brain as well as alterations in the gut microbiome composition might contribute to Alzheimer's disease (AD). Human amyloid precursor protein knock-in (KI) mice contain the Swedish and Iberian mutations (AppNL-F) or those two and also the Arctic mutation (AppNL-G-F). In this study, we assessed whether behavioral and cognitive performance in 6-month-old AppNL-F, AppNL-G-F, and C57BL/6J wild-type (WT) mice was associated with the gut microbiome, and whether the genotype modulates this association. The genotype effects observed in behavioral tests were test-dependent. The biodiversity and composition of the gut microbiome linked to various aspects of mouse behavioral and cognitive performance but differences in genotype modulated these relationships. These genotype-dependent associations include members of the Lachnospiraceae and Ruminococcaceae families. In a subset of female mice, we assessed DNA methylation in the hippocampus and investigated whether alterations in hippocampal DNA methylation were associated with the gut microbiome. Among other differentially methylated regions, we identified a 1 Kb region that overlapped ing 3'UTR of the Tomm40 gene and the promoter region of the Apoe gene that and was significantly more methylated in the hippocampus of AppNL-G-F than WT mice. The integrated gut microbiome hippocampal DNA methylation analysis revealed a positive relationship between amplicon sequence variants (ASVs) within the Lachnospiraceae family and methylation at the Apoe gene. Hence, these microbes may elicit an impact on AD-relevant behavioral and cognitive performance via epigenetic changes in AD-susceptibility genes in neural tissue or that such changes in the epigenome can elicit alterations in intestinal physiology that affect the growth of these taxa in the gut microbiome.

Age-Associated DNA Methylation Patterns Are Shared Between the Hippocampus and Peripheral Blood Cells.

As the population ages, interest in identifying biomarkers of healthy aging and developing antiaging interventions has increased. DNA methylation has emerged as a potentially powerful molecular marker of aging. Methylation changes at specific sites in the human genome that have been identified in peripheral blood have been used as robust estimators of chronological age. Similar age-related DNA methylation signatures are also seen in various tissue types in rodents. However, whether these peripheral alterations in methylation status reflect changes that also occur in the central nervous system remains unknown. This study begins to address this issue by identifying age-related methylation patterns in the hippocampus and blood of young and old mice. Reduced-representation bisulfite sequencing (RBSS) was used to identify differentially methylated regions (DMRs) in the blood and hippocampus of 2- and 20-month-old C57/Bl6 mice. Of the thousands of DMRs identified genome-wide only five were both found in gene promoters and significantly changed in the same direction with age in both tissues. We analyzed the hippocampal expression of these five hypermethylated genes and found that three were expressed at significantly lower levels in aged mice [suppressor of fused homolog (Sufu), nitric oxide synthase 1 (Nos1) and tripartite motif containing 2 (Trim2)]. We also identified several transcription factor binding motifs common to both hippocampus and blood that were enriched in the DMRs. Overall, our findings suggest that some age-related methylation changes that occur in the brain are also evident in the blood and could have significant translational relevance.

RNA-Seq of human whole blood: Evaluation of globin RNA depletion on Ribo-Zero library method.

Peripheral blood is a highly accessible biofluid providing a rich source of information about human physiology and health status. However, for studies of the blood transcriptome with RNA sequencing (RNA-Seq) techniques, high levels of hemoglobin mRNAs (hgbRNA) present in blood can occupy valuable sequencing space, impacting detection and quantification of non-hgbRNAs. In this study, we evaluated two methods for preparing ribosomal RNA (rRNA)-depleted sequencing libraries for RNA-Seq of whole blood, one of which is also designed to deplete hgbRNAs. Two experiments were performed: one evaluating library performance across 6 human blood samples and the other examining library reproducibility and performance in a two-subject subset. We find that addition of hgbRNA depletion to the rRNA-depletion protocol for library preparation from blood RNA effectively reduces highly abundant hgbRNA reads; however, it does not result in a statistically significant increase in differentially expressed genes in our patient-control study. Bioinformatic removal of globin gene counts in non-hgbRNA depleted libraries provides improvement in overall performance of these libraries. We conclude that use of a standard ribosomal RNA depletion method for library preparation coupled with bioinformatic removal of globin gene counts is sufficient for reproducible and sensitive measurement of both coding and noncoding RNAs in the blood transcriptome.

Dynamic changes in chromatin accessibility, altered adipogenic gene expression, and total versus de novo fatty acid synthesis in subcutaneous adipose stem cells of normal-weight polycystic ovary syndrome (PCOS) women during adipogenesis: evidence of cellular programming.

BACKGROUND: Normal-weight polycystic ovary syndrome (PCOS) women exhibit adipose resistance in vivo accompanied by enhanced subcutaneous (SC) abdominal adipose stem cell (ASC) development to adipocytes with accelerated lipid accumulation per cell in vitro. The present study examines chromatin accessibility, RNA expression and fatty acid (FA) synthesis during SC abdominal ASC differentiation into adipocytes in vitro of normal-weight PCOS versus age- and body mass index-matched normoandrogenic ovulatory (control) women to study epigenetic/genetic characteristics as well as functional alterations of PCOS and control ASCs during adipogenesis. RESULTS: SC abdominal ASCs from PCOS women versus controls exhibited dynamic chromatin accessibility during adipogenesis, from significantly less chromatin accessibility at day 0 to greater chromatin accessibility by day 12, with enrichment of binding motifs for transcription factors (TFs) of the AP-1 subfamily at days 0, 3, and 12. In PCOS versus control cells, expression of genes governing adipocyte differentiation (PPARγ, CEBPα, AGPAT2) and function (ADIPOQ, FABP4, LPL, PLIN1, SLC2A4) was increased two-sixfold at days 3, 7, and 12, while that involving Wnt signaling (FZD1, SFRP1, and WNT10B) was decreased. Differential gene expression in PCOS cells at these time points involved triacylglycerol synthesis, lipid oxidation, free fatty acid beta-oxidation, and oxidative phosphorylation of the TCA cycle, with TGFB1 as a significant upstream regulator. There was a broad correspondence between increased chromatin accessibility and increased RNA expression of those 12 genes involved in adipocyte differentiation and function, Wnt signaling, as well as genes involved in the triacylglycerol synthesis functional group at day 12 of adipogenesis. Total content and de novo synthesis of myristic (C14:0), palmitic (C16:0), palmitoleic (C16:1), and oleic (C18:1) acid increased from day 7 to day 12 in all cells, with total content and de novo synthesis of FAs significantly greater in PCOS than controls cells at day 12. CONCLUSIONS: In normal-weight PCOS women, dynamic chromatin remodeling of SC abdominal ASCs during adipogenesis may enhance adipogenic gene expression as a programmed mechanism to promote greater fat storage.

Synergistic Effects of Hyperandrogenemia and Obesogenic Western-style Diet on Transcription and DNA Methylation in Visceral Adipose Tissue of Nonhuman Primates.

Polycystic ovary syndrome (PCOS) is a major reproductive disorder that is responsible for 80% of anovulatory infertility and that is associated with hyperandrogenemia, increased risk of obesity, and white adipose tissue (WAT) dysfunction. We have previously demonstrated that the combination of chronic testosterone (T) treatment and an obesogenic Western-style diet (WSD) exerts synergistic functional effects on WAT, leading to increased lipid accumulation in visceral adipocytes by an unknown mechanism. In this study, we examined the whole-genome transcriptional response in visceral WAT to T and WSD, alone and in combination. We observed a synergistic effect of T and WSD on gene expression, resulting in upregulation of lipid storage genes concomitant with adipocyte hypertrophy. Because DNA methylation is known to be associated with body fat distribution and the etiology of PCOS, we conducted whole-genome DNA methylation analysis of visceral WAT. While only a fraction of differentially expressed genes also exhibited differential DNA methylation, in silico analysis showed that differentially methylated regions were enriched in transcription factor binding motifs, suggesting a potential gene regulatory role for these regions. In summary, this study demonstrates that hyperandrogenemia alone does not induce global transcriptional and epigenetic response in young female macaques unless combined with an obesogenic diet.

Corneal refractive changes due to short-term eyelid pressure in downward gaze.

PURPOSE: To assess corneal refractive changes after 15-minute visual tasks and their association with eyelid morphology. SETTING: Contact Lens and Visual Optics Laboratory, School of Optometry, Queensland University of Technology, Brisbane, Queensland, Australia. METHODS: Eighteen young subjects with normal ocular health were recruited. Corneal topography was measured with a videokeratoscope before and after 4 conditions consisting of 2 downward gaze angles (20 degrees and 40 degrees) and 2 types of visual tasks (reading and steady fixation). Anterior eye photography in downward gaze was used to determine the eyelid angle, tilt, and position with respect to the cornea. RESULTS: Corneal refractive power changed significantly after the 15-minute downward gaze tasks. The largest mean corneal spherocylindrical change was +0.33 -0.30 x 84 after reading in the 40-degree downward gaze (4.0 mm corneal diameter). The refractive changes were significantly larger after the 40-degree tasks than after the 20-degree tasks (P<.001). The changes in refractive root-mean-square error were significant for all conditions, except the 20-degree steady fixation task, with 4.0 and 6.0 mm analysis diameters (P<.05). Significant correlations were found between some aspects of eyelid morphometry and corneal refractive change. CONCLUSIONS: The pressure of the eyelids on the cornea in short-term downward gaze resulted in optically and clinically relevant corneal changes. Correlation between the refractive corneal changes and eyelid parameters suggests that the angle, shape, and position of the eyelids influence the nature of the corneal changes. When high accuracy is required, refraction should be qualified by the visual tasks undertaken before assessment.

24-h fluid kinetics and perception of sweat losses following a 1-h run in a temperate environment.

This study examined 24-h post-run hydration status and sweat loss estimation accuracy in college age runners (men=12, women=8) after completing a 1-h self-paced outdoor run (wet bulb globe temperature=19.9±3.0 °C). Sweat losses (1353±422 mL; 1.9%±0.5% of body mass) were significantly greater (p<0.001) than perceived losses (686±586 mL). Cumulative fluid consumption equaled 3876±1133 mL (218±178 mL during) with 37% of fluid ingested lost through urine voids (1450±678 mL). Fluid balance based on intake and urine production equaled +554±669 mL at 12 h and +1186±735 mL at 24 h. Most runners reported euhydrated (pre-run urine specific gravity (USG)=1.018±0.008) with no changes (p=0.33) at hours 12 or 24 when both genders were included. However, USG was higher (p=0.004) at 12 h post-run for men (1.025±0.0070 vs. 1.014±0.007), who consumed 171%±40% of sweat losses at 12 h vs. 268%±88% for women. Most runners do not need intervention concerning between bout hydration needs in temperate environments. However, repeated USG measurements were able to identify runners who greatly under or over consumed fluid during recovery. Practitioners can use multiple USG assessments as cheap method to detect runners who need to modify their hydration strategies and should promote assessment of sweat losses by change in body mass, as runners had poor perception of sweat losses.

Corneal changes following short-term rigid contact lens wear.

PURPOSE: The aim of this cross-over study was to investigate the changes in corneal thickness, anterior and posterior corneal topography, corneal refractive power and ocular wavefront aberrations, following the short term use of rigid contact lenses. METHOD: Fourteen participants wore 4 different types of contact lenses (RGP lenses of 9.5mm and 10.5mm diameter, and for comparison a PMMA lens of 9.5mm diameter and a soft silicone hydrogel lens) on 4 different days for a period of 8h on each day. Measures were collected before and after contact lens wear and additionally on a baseline day. RESULTS: Anterior corneal curvature generally showed a flattening with both of the RGP lenses and a steepening with the PMMA lens. A significant negative correlation was found between the change in corneal swelling and central and peripheral posterior corneal curvature (all p ≤ 0.001). RGP contact lenses caused a significant decrease in corneal refractive power (hyperopic shift) of approximately 0.5D. The PMMA contact lenses caused the greatest corneal swelling in both the central (27.92 ± 15.49 µm, p<0.001) and peripheral (17.78 ± 12.11 µm, p=0.001) corneal regions, a significant flattening of the posterior cornea and an increase in ocular aberrations (all p ≤ 0.05). CONCLUSION: The corneal swelling associated with RGP lenses was relatively minor, but there was slight central corneal flattening and a clinically significant hyperopic change in corneal refractive power after the first day of lens wear. The PMMA contact lenses resulted in significant corneal swelling and reduced optical performance of the cornea.

Axial length changes with shifts of gaze direction in myopes and emmetropes.

PURPOSE: The purpose of the study was to investigate the changes in axial length occurring with shifts in gaze direction. METHODS: Axial length measurements were obtained from the left eye of 30 young adults (10 emmetropes, 10 low myopes, and 10 moderate myopes) through a rotating prism with 15° deviation, along the foveal axis, using a noncontact optical biometer in each of the nine different cardinal directions of gaze over 5 minutes. The subject's fellow eye fixated on an external distance (6 m) target to control accommodation, also with 15° deviation. Axial length measurements were also performed in 15° and 25° downward gaze with the biometer inclined on a tilting table, allowing gaze shifts to be achieved with either full head turn but no eye turn, or full eye turn with no head turn. RESULTS: There was a significant influence of gaze angle and time on axial length (both P < 0.001), with the greatest axial elongation (+18 ± 8 µm) occurring with inferonasal gaze (P < 0.001) and a slight decrease in axial length in superior gaze (-12 ± 17 µm) compared with primary gaze (P < 0.001). In downward gaze, a significant axial elongation occurred when eye turn was used (P < 0.001), but not when head turn was used to shift gaze (P > 0.05). CONCLUSIONS: The angle of gaze has a small but significant short-term effect on axial length, with greatest elongation occurring in inferonasal gaze. The elongation of the eye appears to be due to the influence of the extraocular muscles, in particular the oblique muscles.

Measurement of ocular aberrations in downward gaze using a modified clinical aberrometer.

Changes in corneal optics have been measured after downward gaze. However, ocular aberrations during downward gaze have not been previously measured. A commercial Shack-Hartmann aberrometer (COAS-HD) was modified by adding a relay lens system and a rotatable beam splitter to allow on-axis aberration measurements in primary gaze and downward gaze with binocular fixation. Measurements with the modified aberrometer (COAS-HD relay system) in primary and downward gaze were validated against a conventional aberrometer. In human eyes, there were significant changes (p<0.05) in defocus C(2,0), primary astigmatism C(2,2) and vertical coma C(3,-1) in downward gaze (25 degrees) compared to primary gaze, indicating the potential influence of biomechanical forces on the optics of the eye in downward gaze. To demonstrate a further clinical application of this modified aberrometer, we measured ocular aberrations when wearing a progressive addition lens (PAL) in primary gaze (0 degree), 15 degrees downward gaze and 25 degrees downward gaze.