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Ophidian serpentoviruses, positive-sense RNA viruses in the order Nidovirales , are important infectious agents of both captive and free-ranging reptiles. Although the clinical significance of these viruses can be variable, some serpentoviruses are pathogenic and potentially fatal in captive snakes. While serpentoviral diversity and disease potential are well documented, little is known about the fundamental properties of these viruses, including their potential host ranges, kinetics of growth, environmental stability, and susceptibility to common disinfectants and viricides. To address this, three serpentoviruses were isolated in culture from three unique PCR-positive python species: Ball python ( Python regius ), green tree python ( Morelia viridis ), and Stimson's python ( Antaresia stimsoni ). A median tissue culture infectious dose (TCID 50 ) was established to characterize viral stability, growth, and susceptibility. All isolates showed an environmental stability of 10-12 days at room temperature (20 °C). While all three viruses produced variable peak titers on three different cell lines when incubated at 32 °C, none of the viruses detectably replicated at 35 °C. All viruses demonstrated a wide susceptibility to sanitizers, with 10% bleach, 2% chlorhexidine, and 70% ethanol inactivating the virus in one minute and 7% peroxide and a quaternary ammonium solution within three minutes. Of seven tested antiviral agents, remdesivir, ribavirin, and NITD-008, showed potent antiviral activity against the three viruses. Finally, the three isolates successfully infected 32 unique tissue culture cell lines representing different diverse reptile taxa and select mammals and birds as detected by epifluorescent immunostaining. This study represents the first characterization of in vitro properties of growth, stability, host range, and inactivation for a serpentovirus. The reported results provide the basis for procedures to mitigate the spread of serpentoviruses in captive snake colonies as well as identify potential non-pharmacologic and pharmacologic treatment options for ophidian serpentoviral infections.

Purpose: The lamina cribrosa (LC) is a layer of fenestrated connective tissue tethered to the posterior sclera across the scleral canal in the optic nerve head (ONH). It is located at the interface of intracranial and intraocular compartments and is exposed to intraocular pressure (IOP) anteriorly and intracranial pressure (ICP) or Cerebrospinal fluid (CSF) pressure (CSFP) posteriorly. We hypothesize that the pressure difference across LC will determine LC position and meridional diameter of scleral canal (also called Bruch's membrane opening diameter; BMOD)., Methods: We enrolled 19 human subjects undergoing a medically necessary lumbar puncture (LP) to lower CSFP and 6 anesthetized pigs, whose ICP was increased in 5 mm Hg increments using a lumbar catheter. We imaged ONH using optical coherence tomography and measured IOP and CSFP/ICP at baseline and after each intervention. Radial tomographic ONH scans were analyzed by two independent graders using ImageJ, an open-source software. The following ONH morphological parameters were obtained: BMOD, anterior LC depth and retinal thickness. We modeled effects of acute CSFP/ICP changes on ONH morphological parameters using ANOVA (human study) and generalized linear model (pig study)., Results: For 19 human subjects, CSFP ranged from 5 to 42 mm Hg before LP and 2 to 19.4 mm Hg after LP. For the six pigs, baseline ICP ranged from 1.5 to 9 mm Hg and maximum stable ICP ranged from 18 to 40 mm Hg. Our models showed that acute CSFP/ICP changes had no significant effect on ONH morphological parameters in both humans and pigs., Conclusion: We conclude that ONH does not show measurable morphological changes in response to acute changes of CSFP/ICP. Proposed mechanisms include compensatory and opposing changes in IOP and CSFP/ICP and nonlinear or nonmonotonic effects of IOP and CSFP/ICP across LC.

DNA metabarcoding can provide a high-throughput and rapid method for characterizing responses of communities to environmental stressors. However, within bulk samples, DNA metabarcoding hardly distinguishes live from the dead organisms. Here, both DNA and RNA metabarcoding were applied and compared in experimental freshwater mesocosms conducted for assessment of ecotoxicological responses of zooplankton communities to remediation treatment until 38 days post oil-spill. Furthermore, a novel indicator of normalized vitality (NV), sequence counts of RNA metabarcoding normalized by that of DNA metabarcoding, was developed for assessment of ecological responses. DNA and RNA metabarcoding detected similar taxa richness and rank of relative abundances. Both DNA and RNA metabarcoding demonstrated slight shifts in measured α-diversities in response to treatments. NV presented relatively greater magnitudes of differential responses of community compositions to treatments compared to DNA or RNA metabarcoding. NV declined from the start of the experiment (3 days pre-spill) to the end (38 days post-spill). NV also differed between Rotifer and Arthropoda, possibly due to differential life histories and sizes of organisms. NV could be a useful indicator for characterizing ecological responses to anthropogenic influence; however, the biology of target organisms and subsequent RNA production need to be considered., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Complex chemical reaction environments, such as those found in combustion engines, the upper atmosphere, or the interstellar medium, can contain large numbers of different reactive species participating in similarly large numbers of different chemical reactions. In such settings, identifying the most-likely multistep reaction mechanisms which lead to the production of a particular defined product species is an extremely challenging problem, requiring search and evaluation over a large number of different possible candidate mechanisms while also addressing the permutational challenges posed when considering a large number of reaction routes available to sets of identical molecular species. In this article, the problem of generating candidate reaction mechanisms which form a defined product from a diverse set of reactive molecules is cast as a discrete optimization of a permutationally invariant cost function describing similarity between the target product and the product generated by a trial reaction mechanism. This approach is demonstrated by generating 2230 candidate reaction mechanisms which form benzene from diverse sets of reactive molecules which have been experimentally identified in the interstellar medium. By screening this set of autogenerated mechanisms, using dispersion-corrected DFT to evaluate reaction energies and activation barriers, we identify several candidate barrierless reaction mechanisms (both previously proposed and new) for benzene formation which may operate in the low temperatures found in the interstellar medium and could be investigated further to supplement existing microkinetic models.

Purpose: Orbital veins such as the retinal veins and episcleral veins drain into the cavernous sinus, an intracranial venous structure. We studied the effects of acute intracranial pressure (ICP) elevation on episcleral venous pressure, intraocular pressure and retinal vein diameter in an established non-survival pig model., Methods: In six adult female domestic pigs, we increased ICP in 5 mm Hg increments using saline infusion through a lumbar drain. We measured ICP (using parenchymal pressure monitor), intraocular pressure (using pneumatonometer), episcleral venous pressure (using venomanometer), retinal vein diameter (using OCT images) and arterial blood pressure at each stable ICP increment. The average baseline ICP was 5.4 mm Hg (range 1.5-9 mm Hg) and the maximum stable ICP ranged from 18 to 40 mm Hg. Linear mixed models with random intercepts were used to evaluate the effect of acute ICP increase on outcome variables., Results: With acute ICP elevation, we found loss of retinal venous pulsation and increased episcleral venous pressure, intraocular pressure and retinal vein pressure in all animals. Specifically, acute ICP increase was significantly associated with episcleral venous pressure (β = 0.31; 95% CI 0.14-0.48, p < .001), intraocular pressure (β = 0.37, 95%CI 0.24-0.50; p < .001) and retinal vein diameter (β = 11.29, 95%CI 1.57-21.00; p = .03) after controlling for the effects of arterial blood pressure., Conclusion: We believe that the ophthalmic effects of acute ICP elevation are mediated by increased intracranial venous pressure producing upstream pressure changes within the orbital and retinal veins. These results offer exciting possibilities for the development of non-invasive ophthalmic biomarkers to estimate acute ICP elevations following significant neuro-trauma.

Introduction: Cross-sectional survey of 92 board-certified practicing Midwestern ophthalmologists to determine why prescribing habits favor brand-name drugs over generics and to identify approaches for increasing generic drug utilization., Methods: A survey was sent to members of state ophthalmology societies, private practice groups, and individual ophthalmologists to evaluate basic demographic/practice information, knowledge and opinions on generic drugs, frequency of drug representative visits, understanding of the Food and Drug Administration's process of evaluating generics, knowledge of patients' financial status and preferences, and action items that would increase generic utilization., Results: Three factors increase the likelihood of ophthalmologists switching patients to generic drugs: increased knowledge of (1) generic options, (2) price differences between brand-names and generics, and ((3) patient preference for generics. The following four factors decrease the likelihood of ophthalmologists switching patients to generic drugs: (1) increased disease severity, (2) feeling that patient outcomes may be affected by choice of brand-name versus generic, (3) personal preference for taking a brand-name drug over a generic for their own hypothetical eye disease even if both were free, and (4) increased personal preference for taking a brand-name drug for their hypothetical eye disease., Conclusion: Ophthalmologists should continue to update themselves on generic medication options, become familiar with the price difference of generics versus brand-name drugs for commonly prescribed medications, and seek patients' opinions on generics and correct them when possible to increase generic utilization. In addition, studies evaluating the clinical equivalence of generic drugs relative to brand-name drugs should be performed and may help increase generic utilization.

Background: Glaucoma prevalence and subtype profile vary across different racial and ethnic groups. This study provides a comparative evaluation of differences in aqueous humour dynamics (AHD) and ocular biometrics in healthy Chinese and Caucasian adults of two different age groups., Methods: Data from two independent studies with identical designs were compared. Cohorts included young adults (20-30 years old, 32 Chinese and 39 Caucasians) and older adults (>50 years old, 37 Chinese and 46 Caucasians). Parameters of AHD and ocular biometrics were evaluated. Group comparisons were made by generalised estimating equation methods., Results: Differences in young adult Caucasians compared with similarly aged Chinese were thinner central cornea (-29.27 µm, p<0.001), lower intraocular pressure (IOP) (-2.33 mm Hg, p<0.001), larger anterior chamber volume (ACV) (28.78 µL, p<0.001) and faster uveoscleral outflow rate (Fu) (0.82 µL/min, p<0.001). Differences in older adult Caucasians compared with similarly aged Chinese were slower aqueous flow rate (Fa) (-0.28 µL/min, p=0.042), lower IOP (-1.97 mm Hg, p<0.001) and larger ACV (33.15 µL, p<0.001). Considering all subjects together by race, Caucasian subjects had slower Fa (-0.22 µL/min, p=0.035), thinner corneas (-0.52 µm, p=0.003), lower IOP (-2.11 mm Hg, p<0.001), higher ACV (30.39 µL, p<0.001) and faster Fu (0.63 µL/min, p<0.001)., Conclusion: Differences in AHD and biometrics between Caucasian and Chinese adults include larger ACVs which may contribute to the wider angles reported in Caucasians, and slower Fa rates coupled with faster Fu rates which may contribute to their lower IOP and lower overall risk of glaucoma., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2019. No commercial re-use. See rights and permissions. Published by BMJ.)

Objective: Several attributes of female reproductive history, including age at natural menopause (ANM), have been related to primary open-angle glaucoma (POAG). We assembled 18 previously reported common genetic variants that predict ANM to determine their association with ANM or POAG., Methods: Using data from the Nurses' Health Study (7,143 women), we validated the ANM weighted genetic risk score in relation to self-reported ANM. Subsequently, to assess the relation with POAG, we used data from 2,160 female POAG cases and 29,110 controls in the National Eye Institute Glaucoma Human Genetics Collaboration Heritable Overall Operational Database (NEIGHBORHOOD), which consists of 8 datasets with imputed genotypes to 5.6+ million markers. Associations with POAG were assessed in each dataset, and site-specific results were meta-analyzed using the inverse weighted variance method., Results: The genetic risk score was associated with self-reported ANM (P = 2.2 × 10) and predicted 4.8% of the variance in ANM. The ANM genetic risk score was not associated with POAG (Odds Ratio (OR) = 1.002; 95% Confidence Interval (CI): 0.998, 1.007; P = 0.28). No single genetic variant in the panel achieved nominal association with POAG (P ≥0.20). Compared to the middle 80 percent, there was also no association with the lowest 10 percentile or highest 90 percentile of genetic risk score with POAG (OR = 0.75; 95% CI: 0.47, 1.21; P = 0.23 and OR = 1.10; 95% CI: 0.72, 1.69; P = 0.65, respectively)., Conclusions: A genetic risk score predicting 4.8% of ANM variation was not related to POAG; thus, genetic determinants of ANM are unlikely to explain the previously reported association between the two phenotypes., Competing Interests: Financial disclosure/conflicts of interest: Please see section at the end of the article.

Purpose: This study examines the effects of 2 Rho kinase inhibitors on intraocular pressure (IOP) and aqueous humor dynamics., Methods: IOPs of New Zealand albino rabbits with ocular normotension and cynomolgus macaques (nonhuman primate, NHP) with chronic unilateral laser-induced glaucoma were measured at baseline and periodically after a 9 a.m. dose of H-1152, Y-27632, or vehicle. In a separate group of NHPs, aqueous flow, outflow facility, uveoscleral outflow, and IOP were determined after treatment with Y-27632 or vehicle control., Results: Decreases in IOP were found in rabbits (n = 5) at 6 h after one dose of 2% Y-27632 (29%, P = 0.0002) or 1% H-1152 (35%, P = 0.0001), and in hypertensive eyes of NHPs (n = 7-9) at 3 h after one dose of 2% Y-27632 (35%, P = 0.005) or 1% H-1152 (51%, P = 0.0003). With 2 doses of 1% Y-27632 or vehicle in NHP hypertensive eyes (n = 12), significant drug effects were IOP reduction of 28% (P = 0.05) at 2.5 h after the second dose and increases in aqueous flow (36%; P = 0.013), uveoscleral outflow (59%, P = 0.008), and outflow facility (40%; P = 0.01). In normotensive eyes of the same animals, aqueous flow increased by 21% (P = 0.03). No significant change was found in any of the other parameters., Conclusions: Y-27632 and H-1152 lower IOP in rabbits and hypertensive eyes of NHPs for at least 6 h after single doses. The Y-27632 effect on IOP in hypertensive NHP eyes is caused by increases in outflow facility and uveoscleral outflow. An increase in aqueous humor formation attenuates but does not prevent an IOP decrease.

Glaucoma is a leading cause of blindness worldwide. Primary open-angle glaucoma (POAG) is the most common subtype and is a complex trait with multigenic inheritance. Genome-wide association studies have previously identified a significant association between POAG and the SIX6 locus (rs10483727, odds ratio (OR) = 1.32, p = 3.87×10(-11)). SIX6 plays a role in ocular development and has been associated with the morphology of the optic nerve. We sequenced the SIX6 coding and regulatory regions in 262 POAG cases and 256 controls and identified six nonsynonymous coding variants, including five rare and one common variant, Asn141His (rs33912345), which was associated significantly with POAG (OR = 1.27, p = 4.2×10(-10)) in the NEIGHBOR/GLAUGEN datasets. These variants were tested in an in vivo Danio rerio (zebrafish) complementation assay to evaluate ocular metrics such as eye size and optic nerve structure. Five variants, found primarily in POAG cases, were hypomorphic or null, while the sixth variant, found only in controls, was benign. One variant in the SIX6 enhancer increased expression of SIX6 and disrupted its regulation. Finally, to our knowledge for the first time, we have identified a clinical feature in POAG patients that appears to be dependent upon SIX6 genotype: patients who are homozygous for the SIX6 risk allele (His141) have a statistically thinner retinal nerve fiber layer than patients homozygous for the SIX6 non-risk allele (Asn141). Our results, in combination with previous SIX6 work, lead us to hypothesize that SIX6 risk variants disrupt the development of the neural retina, leading to a reduced number of retinal ganglion cells, thereby increasing the risk of glaucoma-associated vision loss.

Introduction: "Minutes in Motion" (MIM) was a community-based exercise challenge designed to recruit a large number of people to increase their physical activity for 6 weeks. We examined participant characteristics that improved the likelihood of success., Methods: This program challenged community members to engage in 30 minutes of physical activity every day for 6 weeks. Participants were asked to submit the number of minutes they exercised at the halfway point of the challenge and again at the end. Those who participated were eligible for prizes. Participants also were asked to complete voluntary pre- and post-surveys that included questions about usual amount of physical activity, perceived improvement from the exercise, and self-efficacy to exercise., Results: Of the 3505 community members who signed up for MIM, 78% initially participated and 61% met the challenge goal. The program evaluation was completed by 567 participants. As a result of MIM, 41% of subjects reported they were more active, 51% reported weight loss, 44% had improved endurance, and 51% had improved mood. Fifty-eight percent of subjects exercised more per week at the end of the study than they did before, and self-efficacy to exercise increased in 47% of the subjects. Keeping a log, exercising at work, and wearing a pedometer were related to many of these outcomes., Conclusions: A physical activity challenge targeted at a community can recruit a large number of people to increase their minutes in motion. Outcomes can be enhanced with certain recommended participation elements.

Objective: Older adults are at higher risk for developing chronic conditions such as diabetes, heart disease, or arthritis. Despite the aging process, maintaining independence is a major goal for older adults. Functional fitness has been found to be predictive of one's ability to perform necessary everyday activities needed to maintain independence. We conducted functional fitness assessments with community-dwelling older adults and correlated the findings to other participant characteristics., Methods: Participants completed 6 functional fitness tests and a health-screening questionnaire. Test performance was compared across demographic, behavioral, chronic illness, and activities of daily living categories., Results: One hundred sixty nine adults over age 50 completed the tests. Thirty-seven percent performed at or above the population norm on all tests. There was a significant positive correlation between test performance and activities of daily living (r=0.3520, P=0.0001). In multivariate analysis, the best model to predict test performance included education, self-rated health, obesity, diabetes, and activities of daily living., Conclusions: An objective test, such as the one reported here, may be helpful in predicting loss of independence. However, health care providers, using a few questions based on this study's key findings, may be able to screen for patients with poor functional status that are at risk of losing independence.

Objective: Physical inactivity is a health concern in the United States with nearly 70% of the population getting inadequate amounts of exercise. We set out to determine if wearing a pedometer could significantly increase awareness and amount of physical activity among female employees at a large health care setting., Methods: Employees purchased a pedometer, completed a survey, and were encouraged to walk 10,000 steps daily. Eight weeks later, they completed a follow-up survey., Results: Initially, 510 employees initially participated. Results from 400 women are reported. Setting daily step goals, keeping a log of steps walked, and wearing the pedometer all the time were the indicators most likely to predict significant improvements in level of awareness and amount of physical activity, self-efficacy, and other physical improvements (increased energy, ill less often, and weight loss). A majority (71%) indicated they would continue to wear the pedometer after the study ended., Conclusions: Wearing a pedometer is a simple, non-invasive way for women to increase awareness of daily activity and does lead to increased physical activity. Maximum results in improved activity and improvement in health occurred in women who were most compliant with the intervention.

Objective: To describe an outpatient smoking cessation clinic, and what predicts participant success., Methods: In 1999, a 12-week smoking cessation clinic was begun. In 2000, the duration was expanded to include an additional 12 weeks of relapse prevention. Participants completed surveys at the beginning and end of their clinic. All participants, even those who dropped out, were surveyed to determine current status. If they were not reached, they were assumed to be smoking., Results: Eleven clinics have been completed thus far, with 117 people enrolling. Of all participants, 57% were not smoking at their last contact (average length of follow-up was 14 months). Age, baseline smoking rate, and presence of a non-smoking support person were the best predictors of abstinence., Conclusions: A 57% abstinence rate was achieved in this outpatient clinic, serving a mix of rural and non-rural patients.