Evaluation of Antifungal Activity of Endophytic Bacillus spp. and Identification of Secondary Metabolites Produced Against the Phytopathogenic Fungi.

Endophytic bacteria serve as a rich source of diverse antimicrobial compounds. Recently, there has been a growing interest in utilizing endophytic Bacillus spp. as biological agents against phytogenic fungi, owing to their potential to produce a wide range of antimicrobial substances. The objective of this research was to investigate the protective abilities of 15 endophytic Bacillus spp. isolated from previous study from wheat plant, against the phytopathogenic fungi, Fusarium graminearum and Macrophomina phaseolina. A dual culture plate assay was conducted as a preliminary analysis, revealing that 7 out of 15 endophytic Bacillus spp. demonstrated inhibition against one or both of the phytopathogenic fungi used in this study. All seven endophytes were further assessed for the presence of diffusible antifungal metabolites. The cultures were grown in potato dextrose broth for 120 h, and the cell-free supernatant was extracted and analyzed using the cup plate method. The methanolic extract yielded similar results to the dual culture plate analysis, except for WL2-15. Additionally, deformities in the mycelial structure were examined under the light microscope upon exposure to methanolic extract. Furthermore, the analysis and identification of metabolites were carried out via gas chromatography-mass spectrometry of methanolic extract from selected seven endophytic Bacillus spp. The chromatogram revealed the presence of some major peaks such as tridecanoic acid, methyl ester, hydroperoxide, 1-methylbutyl, 9-octadecenamide, (z)-, hexane-1,3,4-triol, 3,5-dimethyl- tetradecanoic acid. To the best of our knowledge, this is the first report of these biocontrol agents in endophytic Bacillus spp. Interestingly, volatile organic compound production was also seen in all the isolates against the phytopathogenic fungi.

Do rainfed production systems have lower environmental impact over irrigated production systems?: On -farm mitigation strategies.

The intensive agriculture practices improved the crop productivity but escalated energy inputs (EI) and carbon foot print (CF) which contributes to global warming. Hence designing productive, profitable crop management practices under different production systems with low environmental impact (EI and CF) is the need of the hour. To identify the practices, quantification of baseline emissions and the major sources of emissions are required. Indian agriculture has diversified crops and production systems but there is dearth of information on both EI and CF of these production systems and crops. Hence the present study was an attempt to find hot spots and identify suitable strategies with high productivity, energy use efficiency (EUE) and carbon use efficiency (CUE). Energy and carbon balance of castor, cotton, chickpea, groundnut, maize, rice (both rainfed and irrigated), wheat, sugarcane (only irrigated), pigeon pea, soybean, sorghum, pearl millet (only rainfed) in different production systems was assessed. Field specific data on different crop management practices as well as grain and biomass yields were considered. Rainfed production systems had lower EI and CF than irrigated system. The nonrenewable sources of energy like fertilizer (64 %), irrigation (78 %), diesel fuel (75 %) and electricity (67 %) are the major source of energy input. Rainfed crops recorded higher CUE over irrigated condition. Adoption of technologies like efficient irrigation strategies (micro irrigation), enhancing fertilizer use efficiency (site specific nutrient management or slow release fertilizer), conservation agriculture (conservation or reduced tillage) rice cultivation methods (SRI or Direct seeded rice) were the mitigation strategies. These results will help policy makers and stake holders in adoption of suitable strategies for sustainable intensification.

Pontine and bilateral cerebellar lesion in osmotic demyelination syndrome associated with uncontrolled type II diabetes mellitus: a case report.

Osmotic demyelination syndrome (ODS) as a result of the hyperosmolar hyperglycemic state is rare and can present with variable neurological manifestation due to lysis of myelin sheath. Case presentation: A 44-year diabetic male presented with complaints of sudden onset, progressive bilateral weakness in lower limbs, and slurring of speech for the past 1.5 months. Cerebellar examination showed a bilaterally impaired finger nose test, dysdiadochokinesia, impaired heel shin test, and an impaired tandem gait. MRI brain (T2 and fluid-attenuated inversion recovery sequences) showed high signal intensity in the central pons and bilateral cerebellum. With a diagnosis of ODS with poorly controlled diabetes, he was treated with insulin, metformin, and supportive measures following which his symptoms subsided gradually. Clinical discussion: A rapid correction of hyponatremia is considered the most common cause of ODS. Variations in plasma glucose levels, a rare cause of ODS, can cause an abrupt osmolality change causing pontine and extrapontine myelinolysis. Prevention of rapid correction of hyponatremia and rapid changes in plasma osmolality in vulnerable patients is the mainstay of treatment. Conclusions: Clinical features, imaging studies, and monitoring of serum osmolality, serum glucose, and electrolytes aid in diagnosis and favorable outcomes for the patient.

Progression of Stargardt Disease as Determined by Fundus Autofluorescence Over a 24-Month Period (ProgStar Report No. 17).

PURPOSE: To estimate the progression rate of atrophic lesions in Stargardt disease derived from fundus autofluorescence (FAF). DESIGN: International, multicenter, prospective cohort study. METHODS: A total of 259 participants aged ≥6 years with disease-causing variants in the ABCA4 gene were enrolled from 9 centers and followed over a 24-month period. FAF images were obtained every 6 months, and areas of definitely decreased autofluorescence (DDAF) and decreased autofluorescence (DAF) were quantified. Progression rates were estimated from linear mixed models with time as the independent variable. RESULTS: A total of 488 study eyes of 259 participants (88.8% with both eyes) were enrolled and images from 432 eyes were followed for 24 months. The overall estimated progression of DDAF was 0.74 mm2/y (95% CI 0.64-0.85, P < .0001) and that of DAF was 0.64 mm2/y (95% CI 0.57-0.71) over a 24-month period in univariate analysis. Growth rates were strongly dependent on baseline lesion area. After square root transformation, the DDAF growth rate was not dependent on baseline lesion radius (P = .11), whereas the DAF growth rate was dependent (P < .0001). Genotype was not found to significantly impact the growth rate of DDAF or DAF lesions. CONCLUSIONS: FAF may serve as a convenient monitoring tool and suitable end point for interventional clinical trials that aim to slow disease progression. DDAF and DAF lesion sizes at baseline are strong predicting factors for lesion area growth and can be partially accounted for by square root transformation.

Occurrence of osmotic demyelination syndrome in diabetes mellitus: A case report and literature review of various etiologies for osmotic demyelination syndrome.

Osmotic demyelination syndrome is a rare condition reported mainly in the case of rapid correction of hyponatremia, but it can occur even in the case of complicated diabetes mellitus either during rapid correction of hyperglycemia or anytime during the complicated diabetes mellitus. We report a case of complicated diabetes mellitus developing osmotic demyelination syndrome. The patient had presented with altered sensorium and seizure, which was initially diagnosed as hyperglycemia, but during his treatment, the magnetic resonance imaging of brain revealed central pontine myelinolysis. Our search on the causes of osmotic demyelination syndrome other than rapid correction of hyponatremia has revealed several other causes like autoimmune liver disease, Sjogren's syndrome and non-Hodgkin's lymphoma in addition to diabetes mellitus.

Delayed myelopathy after organophosphate intoxication: A case report.

Organophosphate-induced delayed neuropathy, a central-distal axonopathy, passes through latent, progressive, static and improvement phases. During the improvement phase, the peripheral nerves regenerate unmasking the spinal cord lesion with myelopathic features. We report a case of a 16-year-old male who developed myelopathy 6 weeks following chlorpyrifos poisoning. He had a motor weakness of 4/5 in bilateral hips and 3/5 in bilateral knees and ankles. Spasticity and exaggerated reflexes with ankle clonus were present in the lower limbs. Sensory and the upper limb motor examinations were all normal. Pertinent blood, cerebrospinal fluid and nerve conduction tests were normal. Magnetic resonance imaging of the spine showed features of cord atrophy. Three months following physiotherapy, his power improved to 5/5 in bilateral knee and hip joints and 4/5 in bilateral ankles with spasticity. Organophosphate-induced delayed neuropathy can present as earlier as 6 weeks with myelopathy. Previous history of organophosphorous exposure is important in myelopathy or peripheral neuropathy.

Calciphylaxis and Ulcerative Lesions of Bilateral Nipple-Areolar Complex Due to Alcoholic Liver Disease.

When faced with eczematous lesions involving the nipple-areolar complex (NAC), Paget's disease is assumed to be the diagnosis, anything else being considered a "Zebra" necessitating its exclusion on pathology. A middle-aged lady presented with synchronous asymmetrical ulceration of bilateral NAC with pleomorphic calcifications on mammography and simultaneous extensive vascular calcification in bilateral breasts that suggested systemic cause. Calciphylaxis is a rare diagnosis occurring usually due to underlying end-stage renal disease or hyperparathyroidism. There are very few case reports of calciphylaxis due to alcoholic liver disease and no cases to the best of our knowledge involving NAC. We report an extremely rare case of breast and NAC calciphylaxis due to alcoholic liver disease, highlighting need to consider benign etiology when bilateral involvement is present.

Mammography Trends in a Tertiary Care Hospital in Nepal.

BACKGROUND: Mammography is an established screening tool for early detection of breast cancer, with several protocols used worldwide. Such screening programs and related data are lacking in less developed countries. We documented and analyzed the mammographic trends at Tribhuvan University Teaching Hospital, a tertiary care referral center, in Kathmandu, Nepal, to develop baseline data which may be helpful in further researches. METHODS: In this descriptive study, imaging findings of consecutive patients who had undergone mammography between July 2016 and March 2018 were reviewed after obtaining ethical clearance from the Institutional Review Committee. Ultrasonography and histopathological examination were done as needed. Demographics, presenting complaints, breast density, Breast Imaging, Reporting, Assessment and Data System category and final diagnosis were recorded and analyzed using appropriate statistical methods. RESULTS: There were more diagnostic mammograms (62%) than screening with mastalgia the most common presenting complaint. Breast density was less in screening group. Overall, there were more benign lesions with incidence of breast cancer being 4.4% more in the diagnostic group. The age range varied from 22 to 86 years, with 15% (n=219) below 40 years age accounting for one-third of the cases of extremely dense breast and one-fourth of the suspicious lesions. Nearly 50% of breast cancers were seen in patients less than 50 years of age. CONCLUSIONS: The study showed greater number of diagnostic than screening mammograms, with malignancies detected more often in the diagnostic group and younger age. Fewer screening studies suggest a lack of breast cancer awareness in our population who seek medical help only when symptomatic.

Breast Arterial Calcifications on Mammography among Patients Attending the Radiology Department in a Tertiary Care Centre: A Descriptive Cross-sectional Study.

INTRODUCTION: Breast arterial calcifications are common mammographic findings which are associated with coronary artery disease. The aim of this study was to find the prevalence of breast arterial calcifications in women presenting for mammography in a tertiary care centre. METHODS: This descriptive cross-sectional study was performed in the Department of Radiology, in a tertiary care hospital after taking ethical clearance, Reference number 352(6-11)E-2, 077/078, data was collected from Syngovia database from March-June 2021 which included 1614 mammograms. Convenience sampling was done and mammograms evaluated for presence of vascular or non-vascular calcification. Further, vascular calcification was graded. Data was entered in Statistical Package for Social Sciences version 25. Point estimate at 95% Confidence Interval was done, and frequency and proportion were calculated. RESULTS: The prevalence of breast arterial calcification was 188 (11.6%) at 95% Confidence Interval (10.03-13.2). The mean age of women included in this study was 48.42±9.55 years with the largest number of patients in the age group 40-49 years, 682 (42.3%), and least in the age group 80-89 years, 3 (0.2%). All patients in the age group 80-89 years, 3 (100%) had vascular calcifications followed by 70-79 years group, 22 (57.5%) and none in patients younger than 30 years. CONCLUSIONS: We found an increase in the number and grade of vascular calcifications in breasts with the patient's age. When present breast arterial calcifications must be mentioned in mammogram report. Identification of such calcifications on mammogram should prompt further screening for atherosclerotic disease.

Paragonimiasis: A missed diagnosis from Nepal.

Paragonimiasis, or Oriental lung fluke is a parasitic infestation seen in Asia, Africa and South America which is spread by the consumption of crabs and crayfish. To date four cases have been reported from Nepal. Here, we report a case of paragonimiasis in a young male from Kathmandu valley who presented with symptoms of fever, on and off for 1 month, shortness of breath and cough. He was found to have pleural effusion. Sputum examination did not reveal acid fast bacilli, However, based on clinical features, he was started on antitubercular treatment. There was initial improvement, but later, he continued to have cough and hemoptysis over the next 2 years and on subsequent High-Resolution Computerized Tomography (HRCT) he was found to have a cavitary lung lesion in the superior segment of lower lobe of left lung. A possibility of aspergillosis was considered for which he underwent a lobectomy. The gross examination of the lung showed a small cavity measuring 5 mm which revealed on histology a parasitic structure with serous glands within it. In addition, there were many foreign body granulomas with ova within them. A diagnosis of paragonimiasis was made and the patient was started on Praziquantal. He recovered well and is currently asymptomatic. We can learn from this case that the signs and symptoms of paragonimiasis mimic that of tuberculosis and the mistaken diagnosis can lead to unnecessary treatment, prolonged morbidity and loss of time and resources.

Current Imaging Trends in COVID-19 Pneumonia.

COVID-19 has rapidly emerged as a pandemic threatening lives and healthcare systems worldwide. With the emergence of the disease in Nepal, all faculties of medicine need to be well prepared to face the challenge. Fortunately, now plenty of research is available to facilitate our preparedness in the war against COVID-19. The reverse transcriptase-polymerase chain reaction is the current gold standard diagnostic test and chest Computed Tomography scan for screening the disease is considered inappropriate by most society recommendations. The Nepal Radiologists' Association has proposed its guidelines which have been endorsed by the Nepal Medical Council. This article aims to summarize the role of imaging focusing on chest X-ray and Computed Tomography scan including the indications, specific findings, and important differentials. Imaging needs to be done taking necessary precautions, to minimize disease transmission, protect health care personnel, and preserve health care system functioning.

Progression of Stargardt Disease as Determined by Fundus Autofluorescence Over a 12-Month Period: ProgStar Report No. 11.

IMPORTANCE: Sensitive outcome measures for disease progression are needed for treatment trials of Stargardt disease. OBJECTIVE: To estimate the progression rate of atrophic lesions in the prospective Natural History of the Progression of Atrophy Secondary to Stargardt Disease (ProgStar) study over a 12-month period. DESIGN, SETTING, AND PARTICIPANTS: This multicenter prospective cohort study was conducted in an international selection of tertiary referral centers from October 21, 2013, to February 15, 2017. Patients who were affected by Stargardt disease, aged 6 years and older at baseline, and harboring disease-causing variants of the ABCA4 gene were enrolled at 9 centers in the United States, United Kingdom, and continental Europe. Data analysis occurred from November 2016 to January 2017. EXPOSURES: Autofluorescence images obtained with a standard protocol were sent to a central reading center, and areas of definitely decreased autofluorescence, questionably decreased autofluorescence, and the total combined area of decreased autofluorescence were outlined and quantified. Progression rates were estimated from linear mixed models with time as the independent variable. MAIN OUTCOMES AND MEASURES: Yearly rate of progression, using the growth of atrophic lesions measured by autofluorescence imaging. RESULTS: A total of 259 study participants (488 eyes; 230 individuals [88.8%] were examined in both eyes) were enrolled (mean [SD] age at first visit, 33.3 [15.1] years; 118 [54.4%] female). Gradable images were available for evaluation for 480 eyes at baseline and 454 eyes after 12 months. At baseline, definitely decreased autofluorescence was present in 306 eyes, and the mean (SD) lesion size was 3.93 (4.37) mm2. The mean total area of decreased autofluorescence at baseline was 4.07 (4.04) mm2. The estimated progression of definitely decreased autofluorescence was 0.76 (95% CI, 0.54-0.97) mm2 per year (P < .001), and the total area of both questionably and definitely decreased autofluorescence was 0.64 (95% CI, 0.50-0.78) mm2 per year (P < .001). Both progression rates depended on initial lesion size. CONCLUSIONS AND RELEVANCE: In Stargardt disease, autofluorescence imaging may serve as a monitoring tool and definitely decreased autofluorescence and total area as outcome measures for interventional clinical trials that aim to slow disease progression. Rates of progression depended mainly on initial lesion size.

Reproducibility of Measurements of Retinal Structural Parameters Using Optical Coherence Tomography in Stargardt Disease.

PURPOSE: To assess the reproducibility of retinal measurements from optical coherence tomography (OCT) in ABCA4-related Stargardt disease (STGD1). METHODS: The international multicenter Progression of Atrophy Secondary to Stargardt Disease (ProgStar) Study enrolled 259 STGD1 patients. OCT images were graded by the study reading center (RC). Semiautomatic segmentation with manual adjustments was used to segment the layers of retinal pigmentation epithelium, outer segments, inner segments (ISs), outer nuclear layer (ONL), inner retina, and the total retina (TR). The images were overlaid to the Early Treatment Diabetic Retinopathy Study (ETDRS) grid. For each layer, the thickness and the intact area of the ETDRS central subfield, inner ring, and outer ring were recorded, respectively. A different set of RC graders regraded 30 independent ProgStar images to evaluate measurement reproducibility. Reproducibility was assessed graphically and using statistics including intraclass correlation (ICC) and relative absolute difference (RAD). RESULTS: Across all layers, measurements of the ETDRS central subfield had low ICC and/or large RAD. The outer-ring region was not fully captured in some images. For inner ring, good reproducibility was observed for intact area in the IS (ICC = 0.99, RAD = 4%), thicknesses of the ONL (ICC = 0.93, RAD = 6%), and TR (ICC = 0.99, RAD = 1%). CONCLUSIONS: STGD1's complex morphology made outer retina segmentation challenging. Measurements of the inner ring, including the intact area of IS (i.e., the ellipsoid zone [EZ]) and ONL and TR thicknesses, had good reproducibility and showed anatomical impairment. TRANSLATIONAL RELEVANCE: ONL and TR thicknesses and the EZ intact area in the ETDRS inner ring hold potential as structural endpoints for STGD1 trials. Structure-function relationships need to be further established.

Sonography of Palpable Breast Lumps in a Tertiary Health Care Centre in Nepal.

BACKGROUND: With a palpable lesion in the breast, the goal is to diagnose malignancy at the earliest. Ultrasonography is used for evaluating symptomatic patients especially those with dense breasts where mammography gives limited information. The objective of this study was to evaluate the sonographic pattern of the palpable breast lumps and correlate with the final pathological diagnosis. METHODS: This was a retrospective study done at our tertiary health care center, from July 2016 to March 2017, including 121 patients presenting to the ultrasound department with complaint of palpable breast lump and whose pathological reports could be followed up. Various sonographic features were studied, sonography and final diagnosis compared. RESULTS: On sonography, about 46% of the cases were benign, 35 % malignant and 18 % indeterminate while tissue diagnosis revealed 63% to be benign, 34% malignant. The most common lesions in each group and sonographic characteristics were evaluated. Of the benign lesions, fibroadenoma was the most common. Most of the indeterminate lesions on sonography were histologically mastitis. We found nearly 58% of the malignant lesions had microlobulated margins. The sensitivity of sonography was 92.9% and specificity 97.5% with diagnostic accuracy 94.8%. CONCLUSIONS: Most of the palpable lumps were benign in our study, most common being fibroadenoma. We had a relatively higher percentage of malignancy which may be due to patients with obviously benign lesions not undergoing tissue diagnosis in our setting. The sonographic features and diagnosis correlated well with the histological diagnosis.

Scotopic Microperimetric Assessment of Rod Function in Stargardt Disease (SMART) Study: Design and Baseline Characteristics (Report No. 1).

PURPOSE: To describe the study design and characteristics at first visit of participants in the longitudinal Scotopic Microperimetric Assessment of Rod Function in Stargardt Disease (SMART) study. METHODS: Scotopic microperimetry (sMP) was performed in one designated study eye in a subset of participants with molecularly proven ABCA4-associated Stargardt disease (STGD1) enrolled in a multicenter natural history study (ProgStar). Study visits were every 6 months over a period ranging from 6 to 24 months, and also included fundus autofluorescence (FAF). RESULTS: SMART enrolled 118 participants (118 eyes). At the first visit of SMART, the mean sensitivity in mesopic microperimetry was 11.48 (±5.05; range 0.00-19.88) dB and in sMP 11.25 (±5.26; 0-19.25) dB. For FAF, all eyes had a lesion of decreased autofluorescence (mean lesion size 3.62 [±3.48; 0.10-21.46] mm2), and a total of 76 eyes (65.5%) had a lesion of definitely decreased autofluorescence with a mean lesion size of 3.46 (±3.60; 0.21-21.46) mm2. CONCLUSIONS: Rod function is impaired in STGD1 and can be assessed by sMP. Testing rod function may serve as a potential outcome measure for future clinical treatment trials. This is evaluated in the SMART study.

Ultrasonographic Elasticity Contrast Index of Palpable Breast Lumps.

INTRODUCTION: Elastrography used in addition to grey scale sonography increases its specificity. Elasticity contrast index (ECI) is based on strain elastrography and being a quantitative parameter, maybe more easy to obtain and reproducible while researches has been done in ECI in thyroid lesions, this is the first study, to the best of our knowledge to evaluate in breast lesions. This study was done to evaluate the diagnostic accuracy of Elasticity Contrast Index (ECI) in differentiating benign from malignant lesions of breast and to determine its cutoff value. METHODS: This is a descriptive cross-sectional study done at tertiary health care centre, which involved retrospective evaluation of data collected from September 2016 to March 2017. Conventional sonography was done followed by elastography on commercially available ultrasound machine. ECI was calculated in thyroid protocol available in the unit. Histopathological diagnosis was obtained for all the lesions and taken as gold standard. RESULTS: A total of 89 breast lumps were evaluated, of which was 61 (69.3%) were benign and 27 (30.7%) malignant on histopathology. Independent t test revealed the average ECI value of benign lesions was 2.48 and malignant 5.1. Receiver operating curve showed ECI value of 3.25 as the cutoff, above which the lesions were malignant. CONCLUSIONS: ECI is a quantitative elastography technique which can be easily used as an adjunct during breast sonography and can increase its specificity for diagnosing a lesion as malignant. This could reduce the number of false positive biopsies.

Progression of Stargardt Disease as Determined by Fundus Autofluorescence in the Retrospective Progression of Stargardt Disease Study (ProgStar Report No. 9).

Importance: Sensitive outcome measures for disease progression are needed for treatment trials of Stargardt disease. Objective: To describe the yearly progression rate of atrophic lesions in the retrospective Progression of Stargardt Disease study. Design, Setting, and Participants: A multicenter retrospective cohort study was conducted at tertiary referral centers in the United States and Europe. A total of 251 patients aged 6 years or older at baseline, harboring disease-causing variants in ABCA4 (OMIM 601691), enrolled in the study from 9 centers between August 2, 2013, and December 12, 2014; of these patients, 215 had at least 2 gradable fundus autofluorescence images with atrophic lesion(s) present in at least 1 eye. Exposures: Areas of definitely decreased autofluorescence (DDAF) and questionably decreased autofluorescence were quantified by a reading center. Progression rates were estimated from linear mixed models with time as the independent variable. Main Outcomes and Measures: Yearly rate of progression using the growth of atrophic lesions measured by fundus autofluorescence. Results: A total of 251 participants (458 study eyes) were enrolled. Images from 386 eyes of 215 participants (126 females and 89 males; mean [SD] age, 29.9 [14.7] years; mean [SD] age of onset of symptoms, 21.9 [13.3] years) showed atrophic lesions present on at least 2 visits and were graded for 2 (156 eyes), 3 (174 eyes), or 4 (57 eyes) visits. A subset of 224 eyes (123 female participants and 101 male participants; mean [SD] age, 33.0 [15.1] years) had areas of DDAF present on at least 2 visits; these eyes were included in the estimation of the progression of the area of DDAF. At the first visit, DDAF was present in 224 eyes (58.0%), with a mean (SD) lesion size of 2.2 (2.7) mm2. The total mean (SD) area of decreased autofluorescence (DDAF and questionably decreased autofluorescence) at first visit was 2.6 (2.8) mm2. Mean progression of DDAF was 0.51 mm2/y (95% CI, 0.42-0.61 mm2/y), and of total decreased fundus autofluorescence was 0.35 mm2/y (95% CI, 0.28-0.43 mm2/y). Rates of progression depended on the initial size of the lesion. Conclusions and Relevance: In Stargardt disease with DDAF lesions, fundus autofluorescence may serve as a monitoring tool for interventional clinical trials that aim to slow disease progression. Rates of progression depended mainly on initial lesion size.

Incidence of Atrophic Lesions in Stargardt Disease in the Progression of Atrophy Secondary to Stargardt Disease (ProgStar) Study: Report No. 5.

Importance: Outcome measures that are sensitive to disease progression are needed as clinical end points for future treatment trials in Stargardt disease. Objective: To examine the incidence of atrophic lesions of the retinal pigment epithelium in patients with Stargardt disease as determined by fundus autofluorescence imaging. Design, Setting, and Participants: In this retrospective multicenter cohort study, 217 patients 6 years and older at baseline at tertiary referral centers in Europe, the United States, and the United Kingdom who were harboring disease-causing variants in the adenosine triphosphate (ATP)-binding cassette subfamily A member 4 (ABCA4) gene and who met the following criteria were enrolled: (1) at least 1 well-demarcated area of atrophy with a minimum diameter of 300 µm, with the total area of all atrophic lesions being less than or equal to 12 mm2 in at least 1 eye at the most recent visit, and (2) fundus autofluorescence images for at least 2 visits with a minimum of 6 months between at least 2 visits. Data were collected between August 22, 2013, and December 12, 2014. Data analysis was performed from March 15, 2015, through January 31, 2017. Exposures: Images were evaluated by staff at a central reading center. Areas of definitely decreased autofluorescence (DDAF) and questionably decreased autofluorescence (QDAF) were outlined and quantified. Lesion-free survival rates were estimated using Kaplan-Meier survival curves. Main Outcomes and Measures: Incidence of atrophic lesions as determined by fundus autofluorescence. Results: The 217 patients (mean [SD] age, 21.8 [13.3] years; 127 female [57.5%]; 148 white [68.2%]) contributed 390 eyes for which the mean (SD) follow-up time was 3.9 (1.6) years (range, 0.7-12.1 years). Among eyes without DDAF at first visit, the median time to develop a DDAF lesion was 4.9 years (95% CI, 4.3-5.6 years). Among eyes without QDAF, the median time to develop a QDAF lesion was 6.3 years (95% CI, 5.6-9.7 years). Eyes with a lesion of DDAF at the first visit were less likely to develop a QDAF lesion compared with eyes without a lesion of DDAF (hazard ratio, 0.19; 95% CI, 0.05-0.70; P = .01). Conclusions and Relevance: An estimated 50% of the eyes without DDAF at first visit will develop the lesion in less than 5 years, suggesting that incidence of DDAF could serve as an outcome measure for treatment trials.

Comparison of Short-Wavelength Reduced-Illuminance and Conventional Autofluorescence Imaging in Stargardt Macular Dystrophy.

PURPOSE: To compare grading results between short-wavelength reduced-illuminance and conventional autofluorescence imaging in Stargardt macular dystrophy. DESIGN: Reliability study. METHODS: setting: Moorfields Eye Hospital, London (United Kingdom). PATIENTS: Eighteen patients (18 eyes) with Stargardt macular dystrophy. OBSERVATION PROCEDURES: A series of 3 fundus autofluorescence images using 3 different acquisition parameters on a custom-patched device were obtained: (1) 25% laser power and total sensitivity 87; (2) 25% laser power and freely adjusted sensitivity; and (3) 100% laser power and freely adjusted total sensitivity (conventional). The total area of 2 hypoautofluorescent lesion types (definitely decreased autofluorescence and poorly demarcated questionably decreased autofluorescence) was measured. MAIN OUTCOME MEASURES: Agreement in grading between the 3 imaging methods was assessed by kappa coefficients (κ) and intraclass correlation coefficients. RESULTS: The mean ± standard deviation area for images acquired with 25% laser power and freely adjusted total sensitivity was 2.04 ± 1.87 mm(2) for definitely decreased autofluorescence (n = 15) and 1.86 ± 2.14 mm(2) for poorly demarcated questionably decreased autofluorescence (n = 12). The intraclass correlation coefficient (95% confidence interval) was 0.964 (0.929, 0.999) for definitely decreased autofluorescence and 0.268 (0.000, 0.730) for poorly demarcated questionably decreased autofluorescence. CONCLUSIONS: Short-wavelength reduced-illuminance and conventional fundus autofluorescence imaging showed good concordance in assessing areas of definitely decreased autofluorescence. However, there was significantly higher variability between imaging modalities for assessing areas of poorly demarcated questionably decreased autofluorescence.