Climate impacts of landfill gas emissions: Analysis for 20-year and 100-year time horizons.

Climate impacts of landfill gas emissions were investigated for 20- and 100-year time horizons to identify the effects of atmospheric lifetimes of short- and long-lived drivers. Direct and indirect climate impacts were determined for methane and 79 trace species. The impacts were quantified using global warming potential, GWP (direct and indirect); atmospheric degradation (direct); tropospheric ozone forming potential (indirect); secondary aerosol forming potential (indirect) and stratospheric ozone depleting potential (indirect). Effects of cover characteristics, landfill operational conditions, and season on emissions were assessed. Analysis was conducted at five operating municipal solid waste landfills in California, which collectively contained 13% of the waste in place in the state. Climate impacts were determined to be primarily due to direct emissions (99.5 to 115%) with indirect emissions contributing -15 to 0.5%. Methane emissions were 35 to 99% of the total emissions and the remainder mainly greenhouse gases (hydro)chlorofluorocarbons (up to 42% of total emissions) and nitrous oxide. Cover types affected emissions, where the highest emissions were generally from intermediate covers with the largest relative landfill surface areas. Landfill-specific direct emissions varied between 683 and 103,411 and between 381 and 37,925 Mg CO2-eq./yr for 20- and 100-yr time horizons, respectively. Total emissions (direct + indirect) were 680 to 103,600 (20-yr) and were 374 to 38,108 (100-yr) Mg CO2-eq./yr. Analysis time horizon significantly affected emissions. The 20-yr direct and total emissions were consistently higher than the 100-yr emissions by up to 2.5 times. Detailed analysis of time-dependent climate effects can inform strategies to mitigate climate change impacts of landfill gas emissions.

The effect of perinatal high-dose erythropoietin on retinal structural and vascular characteristics in children born preterm.

PURPOSE: To study the long-term effects of perinatal high-dose recombinant human erythropoietin (rhEPO) on macular structural and vascular development in preterm children. DESIGN: Randomized, double-blind clinical trial follow-up plus cohort study. METHODS: Setting: Department of Ophthalmology, University Hospital Zurich, Zurich, Switzerland. STUDY POPULATION: extremely or very preterm born children aged 7-15 years from an ongoing neuropediatric study (EpoKids). These had been previously randomized to receive either high-dose rhEPO or placebo perinatally. INCLUSION CRITERIA: participation in the EpoKids Study, written informed consent (IC). EXCLUSION CRITERIA: previous ocular trauma or surgery; retinal or developmental disease unrelated to prematurity. Term-born children of comparable age were enrolled as a healthy control (HC) group. INCLUSION CRITERIA: term birth, IC. EXCLUSION CRITERIA: any ocular or visual abnormality, high refractive error. Examiners were blinded regarding intervention status until completion of all analyses. (Participants/guardians remain blinded). OBSERVATION PROCEDURES: Spectral-domain OCT scans (Heidelberg Spectralis system) and OCTA imaging (Zeiss PlexElite 9000) were obtained. Ophthalmological and orthoptic examinations excluded ocular comorbidities. MAIN OUTCOME MEASURES: OCT (central retinal thickness, CRT; total macular volume, TMV), superficial plexus OCTA (foveal avascular zone, FAZ; vessel density, VD; vessel length density, VLD) parameters and foveal hypoplasia grade according to published criteria. RESULTS: Macular vessel density parameters (VD and VLD) were significantly lower (p =0.015, CI-95: 0.01 to 0.06 and p=0.015, CI-95: 0.74 to 3.64) in the EPO group (n= 52) when compared to placebo (n=35). No other significant differences were observed between the EPO and placebo group. When comparing the intervention subgroups to HC we found six significant differences in OCT and OCTA parameters (FAZ, VD, VLD and CRT comparing HC and EPO group; FAZ and CRT when comparing HC and placebo group). CONCLUSIONS: Early high-dose rhEPO in infants born extremely or very preterm affects macular vessel density parameters compared to placebo. Premature birth (regardless of intervention status) affects retinal structure and vascular development. Our findings on macular vascular development do not contraindicate the administration of early high-dose EPO in preterm infants. For further understanding of the role of EPO on macular development and its clinical significance, future studies are needed.

Flicker electroretinogram in preterm infants.

BACKGROUND: Infants born prematurely are at risk of developing retinopathy of prematurity, which is associated with abnormalities in retinal function as measured using electroretinography. The aim of this study was to record non-invasive flicker electroretinograms (ERGs) in preterm infants and compare function of moderate and very or extremely preterm infants. METHODS: In this non-randomized, cross-sectional study, 40 moderate preterm (gestational age (GA) 34 0/7 to 36 6/7 weeks, Group A) and 40 very or extremely preterm infants (GA ≤ 31 weeks, Group B) were recruited for flicker ERG recording through closed eyelids using the RETeval® device and skin electrodes. Group A was tested within the first week of life and Group B between 34th and 37th week postmenstrual age. Flicker stimuli were presented at 28.3 Hz with stimulus levels of 3, 6, 12, 30 and 50 cd•s/m2. Primary endpoints were peak time (ms) and amplitude (µV). RESULTS: Flicker ERGs were recordable in most infants with the highest proportion of reproducible ERGs at 30 cd•s/m2. Amplitudes increased with stronger flicker stimulation, while peak times did not differ significantly between stimulus levels nor groups. Amplitudes were significantly greater in Group B at the strongest stimulus level (Mann-Whitney-U-Test=198.00, Z = 4.097, p = <0.001). CONCLUSIONS: Feasibility of collecting flicker ERG data in most preterm infants was confirmed. We found no evidence of reduced retinal responses to flicker stimuli associated with extreme prematurity. Higher amplitudes in very and extremely preterm infants could indicate acceleration of retinal development following birth, triggered by visual stimulation.

Visual outcome measures in pediatric myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD).

BACKGROUND: Myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD) comprises various age-dependent clinical phenotypes and may be monophasic, multiphasic, or chronic. Optic neuritis (ON) is a common manifestation and frequently appears in combination with other MOGAD phenotypes, particularly in young children. Despite permanent structural damage to the retinal nerve fiber layer (RNFL), children often experience complete visual recovery. AIMS: To analyze the progression and impact of MOGAD on the visual system of pediatric patients independently of the history of ON. METHODS: This retrospective study included children who met specific criteria: myelin oligodendrocyte glycoprotein (MOG) immunoglobulin G (IgG) seropositivity, acute presentation of MOGAD, and written general consent. Main outcome measures were global peripapillary retinal nerve fiber layer (pRNFL) thickness, and near and distance visual acuity, analyzed using descriptive statistics. RESULTS: We identified 10 patients with median age of 7.7 years at first event: 7 patients manifested with acute disseminated encephalomyelitis (ADEM) (with ON 5/7, ADEM only 1/7, with transverse myelitis (TM) 1/7), 2 with isolated ON, and 1 patient with neuromyelitis optica spectrum disorder (NMOSD)-like phenotype with ON. Among ON patients, 5/8 were affected bilaterally, with 3 initially diagnosed with unilateral ON but experiencing subsequent involvement of the fellow eye. None of the patients without previous ON showed a deterioration of visual acuity and, if evaluated, a reduction of the pRNFL. CONCLUSION: Most pediatric MOGAD-ON patients in our cohort presented with acute vison loss and optic disc edema. All patients achieved complete visual recovery, independent of number of relapses or initial visual loss. The pRNFL thickness decreased for several months and stabilized at reduced levels after 12 months in the absence of further relapses. MOGAD may not have subclinical/'silent' effects on the visual system, as visual acuity and pRNFL were not affected in patients without ON.

Vitamin A deficiency retinopathy related to medical interventions in a Swiss cohort: a case series.

AIMS OF THE STUDY: Vitamin A deficiency retinopathy is a potentially blinding disease. In developed countries, vitamin A deficiency due to malnutrition is rare. However, vitamin A deficiency can be caused by malabsorption resulting from bowel resection or medication. In this retrospective study, we present five cases of vitamin A deficiency retinopathy related to malabsorption secondary to medical interventions. METHODS: Electronic charts over a ten-year period (2012-2022) were screened for vitamin A deficiency retinopathy. Only patients with vitamin A deficiency confirmed by laboratory tests were included. Symptoms, medical history, visual acuity, optical coherence tomography, fundus autofluorescence, electrophysiological examination, and vitamin A levels were reviewed. RESULTS: Five eligible cases were identified. Median age was 44.7 years (range 22.2-88.9), median duration of ocular symptoms prior to diagnosis was 14 months, and median visual acuity was 1.0 (range 0.5-1.0, Snellen, decimal). Three patients had a history of bariatric surgery, one patient had a small bowel resection and was on octreotide treatment, and one patient suffered from cystic fibrosis and had a history of small bowel resection and severe hepatopathy. Optical coherence tomography showed various abnormalities, including a reduced interdigitation zone, subretinal drusenoid deposits, and a thinned outer nuclear layer. Electroretinogram findings ranged from abnormal oscillatory potentials to non-recordable rod responses. CONCLUSIONS: Vitamin A deficiency retinopathy can occur following medical interventions associated with malabsorption. In cases of night blindness, vitamin A levels should be measured.

Disaster reconnaissance framework for sustainable post-disaster materials management.

A first foundational assessment is provided for disaster debris reconnaissance that includes identifying tools and techniques for reconnaissance activities, identifying challenges in field reconnaissance, and identifying and developing preliminary guidelines and standards based on advancements from a workshop held in 2022. In this workshop, reconnaissance activities were analyzed in twofold: in relation to post-disaster debris and waste materials and in relation to waste management infrastructure. A four-phase timeline was included to capture the full lifecycle of management activities ranging from collection to temporary storage to final management route: pre-disaster or pre-reconnaissance, post-disaster response (days/weeks), short-term recovery (weeks/months), and long-term recovery (months/years). For successful reconnaissance, objectives of field activities and data collection needs; data types and metrics; and measurement and determination methods need to be identified. A reconnaissance framework, represented using a 3x2x2x4 matrix, is proposed to incorporate data attributes (tools, challenges, guides), reconnaissance attributes (debris, infrastructure; factors, actions), and time attributes (pre-event, response, short-term, long-term). This framework supports field reconnaissance missions and protocols that are longitudinally based and focused on post-disaster waste material and infrastructure metrics that advance sustainable materials management practices. To properly frame and develop effective reconnaissance activities, actions for all data attributes (tools, challenges, guides) are proposed to integrate sustainability and resilience considerations. While existing metrics, tools, methods, standards, and protocols can be adapted for sustainable post-disaster materials management reconnaissance, development of new approaches are needed for addressing unique aspects of disaster debris management.

Functional and Morphological Characteristics of the Retina of Patients with Drusen-like Deposits and Systemic Lupus Erythematosus Treated with Hydroxychloroquine: A Retrospective Study.

PURPOSE: To evaluate the impact of drusen-like deposits (DLD) on retinal layer integrity and retinal function by optical coherence tomography (OCT) and multifocal electroretinography (mfERG) in patients with systemic lupus erythematosus (SLE). METHODS: We identified 66 eyes of 33 SLE patients treated with hydroxychloroquine (HCQ) that were categorized into two groups according to whether DLDs were present (34 eyes, Group One) or absent (32 eyes, Group Two). The groups were matched for age, sex, HCQ treatment duration, daily, and cumulative dosage. OCT (retinal layer thicknesses, central retinal thickness, CRT) and mfERG concentric ring analysis were analyzed and compared. RESULTS: CRT was significantly thicker in Group One compared to Group Two (273.21 ± 3.96 vs. 254.5 ± 7.62) (p = 0.023). Group One also demonstrated an overall thicker retinal pigment epithelium compared to Group Two; however, the other outer retinal layers, outer nuclear layer, and photoreceptor layer were found to be significantly thinner in Group One compared to Group Two. We found no differences in mfERG parameters between the two groups. CONCLUSIONS: DLDs in SLE patients lead to abnormal central retinal layer thickness, which has no measurable impact on cone-mediated retinal function assessed by mfERG.

Rapid onset hydroxychloroquine toxicity.

PURPOSE: Hydroxychloroquine (HCQ) can cause irreversible damage to the retina, especially when taken over longer periods. The American Academy of Ophthalmology recommends a regimen for dosing, screening and monitoring of patients treated with HCQ. We present an unusual case of a rapid development of severe HCQ-associated-retinopathy already after 2 years after commencing HCQ treatment. METHODS: Observational case report. Clinical examination, optical coherence tomography (OCT), fundus autofluorescence imaging (FAF), perimetry, full-field & multifocal electroretinography (ERG) were performed. Ancillary tests included neoplastic and paraneoplastic work-up, vitamin levels and whole exome sequencing, in order to rule out other potential causes of a panretinal degeneration. RESULTS: We report on a 58-year-old woman with rheumatoid arthritis, treated initially with 200mg HCQ daily for 1 year (daily dose 3.6mg/kg), then 400mg daily for 1 year (daily dose 7.2mg/kg), and a cumulative dose of 216 g. Her medical history was otherwise unremarkable. No family history for inherited retinal conditions. She was referred due to a rapid and sudden progressive and severe concentric visual field constriction, two years after commencing HCQ treatment. CONCLUSION: This case of a rapid-onset, severe panretinal degeneration shortly after start of HCQ treatment suggests underlying mechanisms and risk factors for HCQ toxicity in addition to those previously reported, and a potential need for supplementary screening tests to prevent HCQ toxicity. AAO dosing guidelines of 5 MG/KG should be strictly adhered to in patients receiving HCQ therapy.

Application of cavity ring-down spectroscopy and a novel near surface Gaussian plume estimation approach to inverse model landfill methane emissions.

Fugitive methane emissions from municipal solid waste landfills impact global climate change and reliable emissions quantification is of increasing importance. Ground-based cavity ring-down spectrometer (CRDS) measurements were used to determine methane concentrations and isotopic compositions of carbon in CH4. Then, CH4 oxidation through various cover materials was assessed using the Keeling plot method. A novel inverse modeling approach using Gaussian dispersion analysis, termed near-surface Gaussian plume estimation (NSGPE), was developed to predict whole-site landfill methane emissions. The concentration data obtained around the landfill perimeter with the mobile ground-based CRDS were used. Methane concentration data were integrated to parameterize discretized point source emissions from a Gaussian dispersion model. Post-processing algorithms were applied to refine modeling predictions to account for the influence of topographical and meteorological conditions on methane transport. Results indicate spatially resolved and consistent emissions estimates among multiple optimization simulations, with refinements increasing the resolution and spatial trends of emissions. Post-processing algorithms resolve consistent overestimation of emissions commonly observed using conventional Gaussian dispersion models.•Ground-based CRDS used to obtain methane concentration and oxidation data.•Novel inverse Gaussian dispersion modeling approach developed to predict methane emissions from landfills accounting for site-specific topography and meteorology.•Post-processing algorithms refine emissions estimates.

Automated detection of hyperreflective foci in the outer nuclear layer of the retina.

PURPOSE: Hyperreflective foci are poorly understood transient elements seen on optical coherence tomography (OCT) of the retina in both healthy and diseased eyes. Systematic studies may benefit from the development of automated tools that can map and track such foci. The outer nuclear layer (ONL) of the retina is an attractive layer in which to study hyperreflective foci as it has no fixed hyperreflective elements in healthy eyes. In this study, we intended to evaluate whether automated image analysis can identify, quantify and visualize hyperreflective foci in the ONL of the retina. METHODS: This longitudinal exploratory study investigated 14 eyes of seven patients including six patients with optic neuropathy and one with mild non-proliferative diabetic retinopathy. In total, 2596 OCT B-scan were obtained. An image analysis blob detector algorithm was used to detect candidate foci, and a convolutional neural network (CNN) trained on a manually labelled subset of data was then used to select those candidate foci in the ONL that fitted the characteristics of the reference foci best. RESULTS: In the manually labelled data set, the blob detector found 2548 candidate foci, correctly detecting 350 (89%) out of 391 manually labelled reference foci. The accuracy of CNN classifier was assessed by manually splitting the 2548 candidate foci into a training and validation set. On the validation set, the classifier obtained an accuracy of 96.3%, a sensitivity of 88.4% and a specificity of 97.5% (AUC 0.989). CONCLUSION: This study demonstrated that automated image analysis and machine learning methods can be used to successfully identify, quantify and visualize hyperreflective foci in the ONL of the retina on OCT scans.

Flicker electroretinogram in newborn infants.

PURPOSE: To develop and validate a flicker electroretinogram (ERG) protocol in term-born neonates as a potential tool for assessing preterm infants at risk of developing retinopathy of prematurity. METHODS: A custom flicker ERG protocol was developed for use with the hand-held RETeval® electrophysiology device. Feasibility of measuring flicker ERG through closed eyelids and without mydriasis was established in a pilot study enabling optimisation of the test protocol. Following this, healthy term-born neonates (gestational age 37-42 weeks) were recruited at the Neonatology clinic of the University Hospital Zurich. Flicker ERG recordings were performed using proprietary disposable skin electrodes during the first four days of life when the infants were sleeping. Flicker stimuli were presented at 28.3 Hz for a stimulus series at 3, 6, 12, 30, and 50 cd·s/m2, with two measurements at each stimulus level. Results were analysed offline. Flicker ERG peak times and amplitudes were derived from the averaged measurements per stimulus level for each subject. RESULTS: 28 term-born neonates were included in the analysis. All infants tolerated the testing procedure well. Flicker ERG recording was achieved in all subjects with reproducible flicker ERG waveforms for 30 and 50 cd·s/m2 stimuli. Reproducible ERGs were recorded in the majority of infants for the weaker stimuli (with detectable ERGs in 20/28, 25/28, and 27/28 at 3, 6, and 12 cd·s/m2, respectively). Flicker ERG amplitudes increased with increasing stimulus strength, with peak times concurrently decreasing slightly. CONCLUSION: Flicker ERG recording is feasible and reliably recorded in sleeping neonates through closed eyelids using skin electrodes and without mydriasis. Flicker ERG amplitude decreases for lower luminance flicker but remains detectable for 3 cd·s/m2 flicker in the majority of healthy term-born neonates. These data provide a basis to study retinal function in premature infants using this protocol.

Assessment of methane emissions from a California landfill using concurrent experimental, inventory, and modeling approaches.

Methane flux and emissions were obtained at a California landfill concurrently using field measurements, inventory analyses, and modeling. Measured fluxes ranged from -3.7 to 828 g/m2-day and generally decreased from daily to intermediate to final covers. Soil covers with high-plasticity clay had the lowest fluxes. Whole-site emissions ranged from 406 to 47,414 tonnes/year (11,368 to 1,327,592 tonnes CO2-eq./year), and were dominated by intermediate covers with high relative surface area. Emissions estimates from flux chamber tests and California Landfill Methane Inventory Model (CALMIM) with oxidation were similar and low, whereas emissions from aerial measurements and CALMIM without oxidation were similar and high. The inventory analyses provided intermediate emissions and a new Gaussian plume model based on ground cavity ring-down spectrometer measurements provided the highest emissions. The assumptions used and the inherent strengths and limitations of the different approaches resulted in the flux and emissions differences. With varied attributes (experimental/modeling; flux/emissions; whole-site/cover-specific, top-down/bottom-up), the approaches provide envelopes of methane emissions and can be used selectively for the two main purposes of landfill methane emissions analysis: to mechanistically determine the factors that control/limit surface emissions and to provide data for atmospheric methane analysis. To reduce emissions, progression from temporary to permanent cover areas can be accelerated and covers with coarser materials can be amended with plastic fines.

Initial Experience with a COVID-19 Screening Chatbot Before Radiology Appointments.

Guidelines for COVID-19 issued by the Centers for Disease Control and Prevention prompted state and local governments to mandate safety measures for screening high-risk patient populations and for institutions to look for ways to limit human contact when possible. The aim of this study was to determine the feasibility of an automated communication system (chatbot) for COVID-19 screening before patients' radiology appointments and to describe patient experiences with the chatbot. We developed a chatbot for COVID-19 screening before outpatient radiology examination appointments and tested it in a pilot study from July 6 to August 31, 2020. The chatbot assessed the presence of any symptoms, exposure, and recent testing. User experience was assessed via a questionnaire based on a 5-point Likert scale. Multivariable logistic regression was performed to predict response rate. The chatbot COVID-19 screening SMS message was sent to 4687 patients. Of these patients, 2722 (58.1%) responded. Of the respondents, 46 (1.7%) reported COVID-19 symptoms; 34 (1.2%) had COVID-19 tests scheduled or pending. Of the 1965 nonresponders, authentication failed for 174 (8.8%), 1496 (76.1%) did not engage with the SMS message, and 251 (12.8%) timed out of the chatbot. The mean rating for the chatbot experience was 4.6. In a multivariable logistic regression model predicting response rate, English written-language preference independently predicted response (odds ratio, 2.71 [95% CI, 1.77-2.77]; P = .007). Age (P = 0.57) and sex (P = 0.51) did not predict response rate. SMS-based COVID-19 screening before scheduled radiology appointments was feasible. English written-language preference (not age or sex) was associated with higher response rate.

Multisystem involvement, defective lysosomes and impaired autophagy in a novel rat model of nephropathic cystinosis.

Recessive mutations in the CTNS gene encoding the lysosomal transporter cystinosin cause cystinosis, a lysosomal storage disease leading to kidney failure and multisystem manifestations. A Ctns knockout mouse model recapitulates features of cystinosis, but the delayed onset of kidney manifestations, phenotype variability and strain effects limit its use for mechanistic and drug development studies. To provide a better model for cystinosis, we generated a Ctns knockout rat model using CRISPR/Cas9 technology. The Ctns-/- rats display progressive cystine accumulation and crystal formation in multiple tissues including kidney, liver and thyroid. They show an early onset and progressive loss of urinary solutes, indicating generalized proximal tubule dysfunction, with development of typical swan-neck lesions, tubulointerstitial fibrosis and kidney failure, and decreased survival. The Ctns-/- rats also present crystals in the cornea, and bone and liver defects, as observed in patients. Mechanistically, the loss of cystinosin induces a phenotype switch associating abnormal proliferation and dedifferentiation, loss of apical receptors and transporters, and defective lysosomal activity and autophagy in the cells. Primary cultures of proximal tubule cells derived from the Ctns-/- rat kidneys confirmed the key changes caused by cystine overload, including reduced endocytic uptake, increased proliferation and defective lysosomal dynamics and autophagy. The novel Ctns-/- rat model and derived proximal tubule cell system provide invaluable tools to investigate the pathogenesis of cystinosis and to accelerate drug discovery.

A three-year longitudinal study of retinal function and structure in patients with multiple sclerosis.

BACKGROUND: Researchers have in recent years begun to investigate ophthalmological manifestations of multiple sclerosis (MS) other than optic neuritis (ON), and it is now clear that changes to retinal function (measured using the electroretinogram, ERG) and structure (measured using optical coherence tomography, OCT) are found in MS patients irrespective of prior ON episodes. ERG results are consistent with dysfunctional bipolar cells, as in other autoimmune diseases. To date, studies have presented only cross-sectional data regarding ERG and OCT. We, therefore, studied the longitudinal course of ERG and OCT in patients with MS, as well as the effect of disability changes and non-ON clinical relapses on these functional and structural measures. METHODS: MS patients (n = 23) participating in an ongoing longitudinal observational study were invited to take part in a 3-year ophthalmological substudy. ERG and OCT were performed, and measures of MS-related disability and relapse history were obtained. Study visits were repeated annually. ERG peak times, rod b-wave amplitude, mixed rod/cone and cone b-/a-wave amplitude ratios, thickness of the peripapillary retinal nerve fibre layer, and volumes of the segmented retinal layers/complexes were analysed. Using generalised estimating equation models adjusted for age, ON, and MS treatment status, we assessed changes to ERG and OCT over the study duration, the effect of changes in disability and recent non-ON MS relapses on ERG and OCT, and the effect of selected OCT parameters on corresponding ERG parameters. RESULTS: At the group level, small fluctuations of several ERG peak times were recorded, with OCT values remaining stable. Increased disability between visits was associated with significant prolongation of mixed rod-cone ERG b-wave peak times. No evidence of associations between OCT and ERG parameters was observed. CONCLUSIONS: Retinal bipolar cell function may be affected by changes in disability in patients with MS; however, recent non-ON MS clinical relapses appear not to affect ERG or OCT results. As ERG changes in MS patients over 3 years are likely to be small and of uncertain clinical relevance, longitudinal studies of retinal function in MS should be planned over an extended period.

Long-Range PCR-Based NGS Applications to Diagnose Mendelian Retinal Diseases.

The purpose of this study was to develop a flexible, cost-efficient, next-generation sequencing (NGS) protocol for genetic testing. Long-range polymerase chain reaction (PCR) amplicons of up to 20 kb in size were designed to amplify entire genomic regions for a panel (n = 35) of inherited retinal disease (IRD)-associated loci. Amplicons were pooled and sequenced by NGS. The analysis was applied to 227 probands diagnosed with IRD: (A) 108 previously molecularly diagnosed, (B) 94 without previous genetic testing, and (C) 25 undiagnosed after whole-exome sequencing (WES). The method was validated with 100% sensitivity on cohort A. Long-range PCR-based sequencing revealed likely causative variant(s) in 51% and 24% of proband from cohorts B and C, respectively. Breakpoints of 3 copy number variants (CNVs) could be characterized. Long-range PCR libraries spike-in extended coverage of WES. Read phasing confirmed compound heterozygosity in 5 probands. The proposed sequencing protocol provided deep coverage of the entire gene, including intronic and promoter regions. Our method can be used (i) as a first-tier assay to reduce genetic testing costs, (ii) to elucidate missing heritability cases, (iii) to characterize breakpoints of CNVs at nucleotide resolution, (iv) to extend WES data to non-coding regions by spiking-in long-range PCR libraries, and (v) to help with phasing of candidate variants.

Genotype-phenotype spectrum in isolated and syndromic nanophthalmos.

PURPOSE: To (i) describe a series of patients with isolated or syndromic nanophthalmos with the underlying genetic causes, including novel pathogenic variants and their functional characterization and (ii) to study the association of retinal dystrophy in patients with MFRP variants, based on a detailed literature review of genotype-phenotype correlations. METHODS: Patients with nanophthalmos and available family members received a comprehensive ophthalmological examination. Genetic analysis was based on whole-exome sequencing and variant calling in core genes including MFRP, BEST1, TMEM98, PRSS56, CRB1, GJA1, C1QTNF5, MYRF and FAM111A. A minigene assay was performed for functional characterization of a splice site variant. RESULTS: Seven patients, aged between three and 65 years, from five unrelated families were included. Novel pathogenic variants in MFRP (c.497C>T, c.899-3C>A, c.1180G>A), and PRSS56 (c.1202C>A), and a recurrent de novo variant in FAM111A (c.1706G>A) in a patient with Kenny-Caffey syndrome type 2, were identified. In addition, we report co-inheritance of MFRP-related nanophthalmos and ADAR-related Aicardi-Goutières syndrome. CONCLUSION: Nanophthalmos is a genetically heterogeneous condition, and the severity of ocular manifestations appears not to correlate with variants in a specific gene. However, retinal dystrophy is only observed in patients harbouring pathogenic MFRP variants. Furthermore, heterozygous carriers of MFRP and PRSS56 should be screened for the presence of high hyperopia. Identifying nanophthalmos as an isolated condition or as part of a syndrome has implications for counselling and can accelerate the interdisciplinary care of patients.

Assessment of Regional Methane Emission Inventories through Airborne Quantification in the San Francisco Bay Area.

This study derives methane emission rates from 92 airborne observations collected over 23 facilities including 5 refineries, 10 landfills, 4 wastewater treatment plants (POTWs), 2 composting operations, and 2 dairies in the San Francisco Bay Area. Emission rates are measured using an airborne mass-balance technique from a low-flying aircraft. Annual measurement-based sectorwide methane emissions are 19,000 ± 2300 Mg for refineries, 136,700 ± 25,900 Mg for landfills, 11,900 ± 1,500 Mg for POTWs, and 11,100 ± 3,400 Mg for composting. The average of measured emissions for each refinery ranges from 4 to 23 times larger than the corresponding emissions reported to regulatory agencies, while measurement-derived landfill and POTW estimates are approximately twice the current inventory estimates. Significant methane emissions at composting facilities indicate that a California mandate to divert organics from landfills to composting may not be an effective measure for mitigating methane emissions unless best management practices are instituted at composting facilities. Complementary evidence from airborne remote sensing imagery indicates atmospheric venting from refinery hydrogen plants, landfill working surfaces, composting stockpiles, etc., to be among the specific source types responsible for the observed discrepancies. This work highlights the value of multiple measurement approaches to accurately estimate facility-scale methane emissions and perform source attribution at subfacility scales to guide and verify effective mitigation policy and action.