Genetic modification of flavone biosynthesis in rice enhances biofilm formation of soil diazotrophic bacteria and biological nitrogen fixation.

Improving biological nitrogen fixation (BNF) in cereal crops is a long-sought objective; however, no successful modification of cereal crops showing increased BNF has been reported. Here, we described a novel approach in which rice plants were modified to increase the production of compounds that stimulated biofilm formation in soil diazotrophic bacteria, promoted bacterial colonization of plant tissues and improved BNF with increased grain yield at limiting soil nitrogen contents. We first used a chemical screening to identify plant-produced compounds that induced biofilm formation in nitrogen-fixing bacteria and demonstrated that apigenin and other flavones induced BNF. We then used CRISPR-based gene editing targeting apigenin breakdown in rice, increasing plant apigenin contents and apigenin root exudation. When grown at limiting soil nitrogen conditions, modified rice plants displayed increased grain yield. Biofilm production also modified the root microbiome structure, favouring the enrichment of diazotrophic bacteria recruitment. Our results support the manipulation of the flavone biosynthetic pathway as a feasible strategy for the induction of biological nitrogen fixation in cereals and a reduction in the use of inorganic nitrogen fertilizers.

Nitrogen fixation in a landrace of maize is supported by a mucilage-associated diazotrophic microbiota.

Plants are associated with a complex microbiota that contributes to nutrient acquisition, plant growth, and plant defense. Nitrogen-fixing microbial associations are efficient and well characterized in legumes but are limited in cereals, including maize. We studied an indigenous landrace of maize grown in nitrogen-depleted soils in the Sierra Mixe region of Oaxaca, Mexico. This landrace is characterized by the extensive development of aerial roots that secrete a carbohydrate-rich mucilage. Analysis of the mucilage microbiota indicated that it was enriched in taxa for which many known species are diazotrophic, was enriched for homologs of genes encoding nitrogenase subunits, and harbored active nitrogenase activity as assessed by acetylene reduction and 15N2 incorporation assays. Field experiments in Sierra Mixe using 15N natural abundance or 15N-enrichment assessments over 5 years indicated that atmospheric nitrogen fixation contributed 29%-82% of the nitrogen nutrition of Sierra Mixe maize.

African Orphan Crops Consortium (AOCC): status of developing genomic resources for African orphan crops.

MAIN CONCLUSION: The African Orphan Crops Consortium (AOCC) successfully initiated the ambitious genome sequencing project of 101 African orphan crops/trees with 6 genomes sequenced, 6 near completion, and 20 currently in progress. Addressing stunting, malnutrition, and hidden hunger through nutritious, economic, and resilient agri-food system is one of the major agricultural challenges of this century. As sub-Saharan Africa harbors a large portion of the severely malnourished population, the African Orphan Crops Consortium (AOCC) was established in 2011 with an aim to reduce stunting and malnutrition by providing nutritional security through improving locally adapted nutritious, but neglected, under-researched or orphan African food crops. Foods from these indigenous or naturalized crops and trees are rich in minerals, vitamins, and antioxidant, and are an integral part of the dietary portfolio and cultural, social, and economic milieu of African farmers. Through stakeholder consultations supported by the African Union, 101 African orphan and under-researched crop species were prioritized to mainstream into African agri-food systems. The AOCC, through a network of international-regional-public-private partnerships and collaborations, is generating genomic resources of three types, i.e., reference genome sequence, transcriptome sequence, and re-sequencing 100 accessions/species, using next-generation sequencing (NGS) technology. Furthermore, the University of California Davis African Plant Breeding Academy under the AOCC banner is training 150 lead African scientists to breed high yielding, nutritious, and climate-resilient (biotic and abiotic stress tolerant) crop varieties that meet African farmer and consumer needs. To date, one or more forms of sequence data have been produced for 60 crops. Reference genome sequences for six species have already been published, 6 are almost near completion, and 19 are in progress.

Arsenic from community water fluoridation: quantifying the effect.

Community water fluoridation is a WHO recommended strategy to prevent dental carries. One debated concern is that hydrofluorosilicic acid, used to fluoridate water, contains arsenic and poses a health risk. This study was undertaken to determine if fluoridation contributes to arsenic in drinking water, to estimate the amount of additional arsenic associated with fluoridation, and compare this to the National Sanitation Foundation/American National Standards Institute (NSF/ANSI) standard and estimates from other researchers. Using surveillance data from Ontario drinking water systems, mixed effects linear regression was performed to examine the effect of fluoridation status on the difference in arsenic concentration between raw water and treated water samples. On average, drinking water treatment was found to reduce arsenic levels in water in both fluoridated and non-fluoridated systems by 0.2 µg/L. However, fluoridated systems were associated with an additional 0.078 µg/L (95% CI 0.021, 0.136) of arsenic in water when compared to non-fluoridated systems (P = 0.008) while controlling for raw water arsenic concentrations, types of treatment processes, and source water type. Our estimate is consistent with concentrations expected from other research and is less than 10% of the NSF/ANSI standard of 1 µg/L arsenic in water. This study provides further information to inform decision-making regarding community water fluoridation.

Light depolarization measurements in malaria: A new job for an old friend.

The use of flow cytometry in malaria research has increased over the last decade. Most approaches use nucleic acid stains to detect parasite DNA and RNA and require complex multi-color, multi-parameter analysis to reliably detect infected red blood cells (iRBCs). We recently described a novel and simpler approach to parasite detection based on flow cytometric measurement of scattered light depolarization caused by hemozoin (Hz), a pigment formed by parasite digestion of hemoglobin in iRBCs. Depolarization measurement by flow cytometry was described in 1987; however, patent issues restricted its use to a single manufacturer's hematology analyzers until 2009. Although we recently demonstrated that depolarization measurement of Hz, easily implemented on a bench top flow cytometer (Cyflow), provided useful information for malaria work, doubts regarding its application and utility remain in both the flow cytometry and malaria communities, at least in part because instrument manufacturers do not offer the option of measuring depolarized scatter. Under such circumstances, providing other researchers with guidance as to how to do this seemed to offer the most expeditious way to resolve the issue. We accordingly examined how several commercially available flow cytometers (CyFlow SL, MoFLo, Attune and Accuri C6) could be modified to detect depolarization due to the presence of free Hz on solution, or of Hz in leukocytes or erythrocytes from rodent or human blood. All were readily adapted, with substantially equivalent results obtained with lasers emitting over a wide wavelength range. Other instruments now available may also be modifiable for Hz measurement. Cytometric detection of Hz using depolarization is useful to study different aspects of malaria. Adding additional parameters, such as DNA content and base composition and RNA content, can demonstrably provide improved accuracy and sensitivity of parasite detection and characterization, allowing malaria researchers and eventually clinicians to benefit from cytometric technology.

Neutralization of vascular endothelial growth factor slows progression of retinal nonperfusion in patients with diabetic macular edema.

OBJECTIVE: To determine the effect of suppression of vascular endothelial growth factor (VEGF) by monthly injection of ranibizumab on posterior retinal nonperfusion (RNP) in patients with diabetic macular edema (DME). DESIGN: Unplanned retrospective analysis of prospectively collected data from 2 randomized, sham injection-controlled, double-masked, multicenter clinical trials. PARTICIPANTS: Six hundred sixty-six patients with DME. METHODS: An independent reading center measured the area of RNP on fluorescein angiograms obtained in the phase 3 RISE and RIDE trials. MAIN OUTCOME MEASURES: The percentage of patients with no posterior RNP. RESULTS: The percentage of patients with no posterior RNP decreased in the sham group between baseline and month 24, but remained relatively stable in the 2 ranibizumab groups. After month 24, the sham group crossed over to receive monthly injections of ranibizumab 0.5 mg, and the differences between the sham and ranibizumab groups were reduced. The percentage of patients who showed an increase in posterior RNP from baseline increased over time in all 3 groups, but at a faster rate in the sham group, resulting in statistically significant differences at every time point between months 3 (9.6% vs. 18.5%; P = 0.016) and 24 (16.1% vs. 37.6%; P<0.0001) for ranibizumab 0.5 mg versus sham and from months 6 (12.3% vs. 23.0%; P = 0.013) through 24 (15.0% vs. 37.6%; P<0.0001) for ranibizumab 0.3 mg. Initiation of ranibizumab in the sham group at month 24 was followed by reduction in the percentage of patients with an increase in posterior RNP from baseline at months 30 and 36, whereas the 2 ranibizumab groups continued their gradual rise. CONCLUSIONS: Just as high VEGF levels contribute to progression of retinal nonperfusion in retinal vein occlusion, the same is true in patients with DME, suggesting that regardless of the underlying disease process, high levels of VEGF can cause closure of retinal vessels. However, our data also suggest that VEGF-induced worsening of retinal perfusion in DME is superimposed on another cause of more gradually worsening perfusion, possibly glucotoxicity. Thus, monthly injections of ranibizumab can slow, but not completely prevent, retinal capillary closure in patients with DME.

De-epithelialized fasciocutaneous turnover flap for recurrent calcaneal wound with osteomyelitis.

Recurrent ulcerations of the foot and ankle almost always present a challenge to lower extremity surgeons. Recalcitrant heel ulcerations with osteomyelitis are especially difficult to treat because of the lack of soft tissue coverage. The turnover flap is a simple, fast, and effective treatment method for lower extremity wounds. It is a de-epithelialized fasciocutaneous flap harvested from the adjacent area of the wound. We believe it is an underused technique for advanced wound closure in the lower extremity. It offers several advantages compared with traditional, more difficult to perform, flaps. We have seen an excellent result 18 months after using the turnover flap in a patient with recurrent posterior heel ulceration with calcaneal osteomyelitis.

Flow Cytometry: The Glass Is Half Full.

Accompanied by a historical perspective of the field of cytometry, this introductory chapter provides a broad view of what flow cytometry can do; hence, the glass is half full.

Ranibizumab treatment outcomes in phakic versus pseudophakic eyes: an individual patient data analysis of 2 phase 3 trials.

OBJECTIVE: To compare visual outcomes in phakic and pseudophakic eyes treated with monthly intravitreal ranibizumab for exudative age-related macular degeneration (AMD). DESIGN: Meta-analysis of individual patient data from 2 phase 3 clinical trials of intravitreal ranibizumab in neovascular AMD (Anti-VEGF Antibody for the Treatment of Predominantly Classic Choroidal Neovascularization in Age-Related Macular Degeneration [ANCHOR], ClinicalTrials.gov number, NCT00061594; and Minimally Classic/Occult Trial of the Anti-VEGF Antibody Ranibizumab in the Treatment of Neovascular Age-Related Macular Degeneration [MARINA], ClinicalTrials.gov number NCT00056836). PARTICIPANTS AND CONTROLS: A total of 1137 patients from 2 phase 3 clinical trials. METHODS: Phakic and pseudophakic eyes were treated with monthly intravitreal ranibizumab (0.3 mg or 0.5 mg), sham injections plus verteporfin photodynamic therapy (ANCHOR), or sham injections alone (MARINA). MAIN OUTCOME MEASURES: Mean change from baseline in Early Treatment Diabetic Retinopathy Study (ETDRS) visual acuity (VA) and the proportion of patients gaining or losing 15 or more ETDRS letters. RESULTS: After adjusting for baseline covariates, no differences were seen in mean change in VA for phakic versus pseudophakic eyes. Pseudophakic eyes were more likely to lose 15 or more letters of vision than phakic eyes at 12 months, but not at 24 months. CONCLUSIONS: Overall, in this analysis, lens status did not demonstrate an independent influence on mean VA for eyes treated with monthly ranibizumab. It is possible that phakic eyes may be less prone to severe vision loss. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Predictive value in retinal vein occlusions of early versus late or incomplete ranibizumab response defined by optical coherence tomography.

PURPOSE: To determine if optical coherence tomography (OCT) at baseline or month 3 in the Treatment of Macular Edema following Branch Retinal Vein Occlusion: Evaluation of Efficacy and Safety (BRAVO) and Treatment of Macular Edema following Central Retinal Vein Occlusion: Evaluation of Efficacy and Safety (CRUISE) studies provides information that predicts visual outcome. DESIGN: Post hoc analysis from 2 prospective, randomized, controlled clinical trials. PARTICIPANTS: Three hundred ninety-seven patients from the BRAVO study and 392 patients from the CRUISE study. METHODS: Time-domain OCT imaging data were analyzed. MAIN OUTCOME MEASURES: Mean change from baseline best-corrected visual acuity (BCVA) letter score at month 6 and month 12. RESULTS: Among ranibizumab-treated patients, 71.2% (0.3 mg) and 78.5% (0.5 mg) in the CRUISE study and 79.1% (0.3 mg) and 84.7% (0.5 mg) in the BRAVO study had central foveal thickness (CFT) of 250 µm or less at month 3 and therefore were categorized as early ranibizumab responders. Early ranibizumab responders had excellent visual outcomes regardless of ranibizumab dose; mean improvement in BCVA letter score at 6 and 12 months was 15.0 to 16.5 (central retinal vein occlusion [CRVO]) and 17.4 to 19.1 (branch retinal vein occlusion [BRVO]). Late or incomplete ranibizumab responders with CRVO (CFT >250 µm at month 3) did not fare as well as early responders if they were treated with 0.3 mg ranibizumab (month 6, P = 0.012). At month 6, compared with ranibizumab-treated CRVO patients with resolved cystoid macular edema (CME) at month 3, those with persistent CME did worse, on average, and significantly so for 0.5 mg (13.1 vs. 18.6; P = 0.027). At baseline, subretinal fluid (SRF) was present in 57% of patients with CRVO and in 45% of patients with BRVO; its presence did not portend a poor outcome in patients treated with ranibizumab for whom SRF was eliminated in almost all by month 3. CONCLUSIONS: At month 3 of ranibizumab treatment, OCT images provide predictive information for patients with CRVO, but not for those with BRVO. Visual outcome at months 6 and 12 was reduced in 0.5 mg ranibizumab-treated patients with CRVO who had persistent CME at month 3. It also was reduced in CRVO for those with CFT of more than 250 µm at month 3 who were treated with 0.3 mg ranibizumab. The findings suggest that late or incomplete responders may need careful follow-up. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Vascular endothelial growth factor promotes progressive retinal nonperfusion in patients with retinal vein occlusion.

OBJECTIVE: Central retinal vein occlusion (CRVO) or branch retinal vein occlusion (BRVO) causes hypoperfusion, high levels of vascular endothelial growth factor (VEGF), macular edema, and loss of vision. Many patients also show areas of complete closure of retinal vessels (retinal nonperfusion [RNP]) that increase over time. The objective was to assess the effect of blocking VEGF on progression of RNP. DESIGN: Retrospective analysis of prospectively collected data from 2 randomized, sham injection-controlled, double-masked, multicenter clinical trials. PARTICIPANTS: A total of 392 and 397 patients with macular edema due to CRVO or BRVO. METHODS: An independent reading center measured the area of RNP on fluorescein angiograms (FAs) in 2 phase III trials investigating the effect of ranibizumab (RBZ; Lucentis; Genentech, Inc, South San Francisco, CA) in patients with CRVO or BRVO. MAIN OUTCOME MEASURES: The percentage of patients with no posterior RNP at months 0, 3, 6, 9, and 12. RESULTS: There was no difference among treatment groups at baseline, but at the month 6 primary end point the percentage of patients with CRVO and no RNP was significantly greater in the RBZ groups (0.3 mg, 82.0%, P = 0.0092; 0.5 mg, 84.0%, P = 0.0067) versus the sham group (67.0%). Reperfusion of nonperfused retina was rare (1%) in sham-treated patients with CRVO, but occurred in 6% to 8% of patients with CRVO treated with RBZ (30% of those who had RNP and could improve). Results in patients with BRVO mirrored those in patients with CRVO. Crossover to 0.5 mg RBZ from sham at month 6 halted the progression of RNP and resulted in improvement in both CRVO and BRVO. CONCLUSIONS: Treatment with RBZ did not worsen RNP in patients with RVO, but rather reduced its occurrence compared with sham. These data provide an important new insight regarding the pathogenesis of RVO; the initial vein occlusion is a precipitating event that causes baseline ischemia and release of VEGF, which then contributes to progression of RNP and thus worsening of ischemia. Timely, aggressive blockade of VEGF prevents the worsening of RNP, promotes reperfusion, and eliminates a positive feedback loop.

Anatomical measures as predictors of visual outcomes in ranibizumab-treated eyes with neovascular age-related macular degeneration.

PURPOSE: To investigate if anatomical characteristics of eyes undergoing ranibizumab therapy were predictive of best-corrected visual acuity (BCVA) outcomes over 2 years. METHODS: Post hoc analyses of patients with age-related macular degeneration from PIER studies, defined by fundus fluorescein angiography, quantitative optical coherence tomography (OCT), and qualitative OCT, were performed to determine if associations with BCVA outcomes could be found. RESULTS: Ranibizumab-treated subgroups defined by baseline fundus fluorescein angiography lesion size and composition did not differ in BCVA outcomes at month 24 (P = 0.13-1.0). Inactivity on fundus fluorescein angiography at month 3 was associated with a 12-letter gain by month 12 (P < 0.01), whereas inactivity on month 3 qualitative OCT was not (P > 0.05). Qualitative OCT inactivity at month 5 and separately at month 8 was associated with greater BCVA gains by month 24 (7.1 and 9.5 letters, respectively; P ≤ 0.045) versus eyes with OCT activity. CONCLUSION: When assessed separately, eyes with qualitative OCT (Months 5 and 8) or fundus fluorescein angiography (months 3 and 5) inactivity maintained vision gain from baseline at month 24, while those with leakage not only lost initial vision gains achieved by intraocular ranibizumab but also had net vision losses from baseline at month 24. The PIER infrequent dosing regimen likely exaggerated and accelerated the deleterious effects of retinal fluid on BCVA, and it is not known whether these findings are applicable to treatment regimens that use more frequent monitoring and dosing of ranibizumab.

Rabies in an imported dog, Ontario, Canada, 2022.

Importation of rabies-infected dogs results in significant and costly public and animal health risks. In January 2022, a dog in Ontario, Canada, which was imported from Iran in June 2021, developed rabies, leading to an extensive public health investigation and administration of rabies post-exposure prophylaxis to 37 individuals. The dog was infected with a rabies virus variant known to circulate in Iran. This is the second reported case of a rabies-infected dog imported into Canada in 2021 from a high-risk country for canine mediated rabies. This case emphasizes the need for public education regarding the risks associated with importing dogs from high-risk countries for canine-mediated rabies and the benefits of establishing a public health team specializing in rabies exposure investigations.

The effect of fellow eye visual acuity on visual acuity of study eyes receiving ranibizumab for age-related macular degeneration.

PURPOSE: To investigate whether extremes in visual acuity (very good or very poor) of the fellow eye (FE) influence visual acuity of the study eye in patients receiving intravitreal ranibizumab treatment for neovascular age-related macular degeneration. METHODS: From 2 randomized, controlled, clinical trials (MARINA and ANCHOR), we performed a retrospective analysis of ranibizumab-treated patients who maintained stable FE visual acuity (±5 letters from baseline at each of Months 1, 4, 6, and 12), comparing patients with untreated FE visual acuity that was either 20/32 or better (very good) or 20/200 or worse (very poor). Visual acuity of the treated study eyes, which received monthly intravitreal ranibizumab (0.3 mg or 0.5 mg), was compared between the 2 FE cohorts at the Month 6 and Month 12 visits. RESULTS: A total of 145 patients were analyzed. In the cohort with very poor FE visual acuity (n = 55), there were 35 patients in MARINA and 20 patients in ANCHOR; in the cohort with very good FE visual acuity (n = 90), there were 52 patients in MARINA and 38 patients in ANCHOR.The mean (standard deviation) gain of the study eye visual acuity in the very good FE cohort was 10.3 (13.3) and 10.8 (13.7) letters at Months 6 and 12, respectively, compared with a lesser mean visual acuity gain of 4.6 (12.2) and 6.7 (11.7) letters at Months 6 and 12 in the very poor vision FE cohort. There was no statistically significant difference (adjusted) in the study eye visual acuity change between the 2 cohorts at either 6 months (P = 0.11) or 12 months (P = 0.26). CONCLUSION: This retrospective analysis of the MARINA and ANCHOR study data did not support the hypothesis that FE visual acuity plays a significant role in driving visual acuity of patients receiving monthly intravitreal ranibizumab injections for neovascular age-related macular degeneration. Visual acuity of the FE by itself is, therefore, not a useful parameter in predicting visual acuity in a majority of ranibizumab-treated patients.

A genetically anchored physical framework for Theobroma cacao cv. Matina 1-6.

BACKGROUND: The fermented dried seeds of Theobroma cacao (cacao tree) are the main ingredient in chocolate. World cocoa production was estimated to be 3 million tons in 2010 with an annual estimated average growth rate of 2.2%. The cacao bean production industry is currently under threat from a rise in fungal diseases including black pod, frosty pod, and witches' broom. In order to address these issues, genome-sequencing efforts have been initiated recently to facilitate identification of genetic markers and genes that could be utilized to accelerate the release of robust T. cacao cultivars. However, problems inherent with assembly and resolution of distal regions of complex eukaryotic genomes, such as gaps, chimeric joins, and unresolvable repeat-induced compressions, have been unavoidably encountered with the sequencing strategies selected. RESULTS: Here, we describe the construction of a BAC-based integrated genetic-physical map of the T. cacao cultivar Matina 1-6 which is designed to augment and enhance these sequencing efforts. Three BAC libraries, each comprised of 10× coverage, were constructed and fingerprinted. 230 genetic markers from a high-resolution genetic recombination map and 96 Arabidopsis-derived conserved ortholog set (COS) II markers were anchored using pooled overgo hybridization. A dense tile path consisting of 29,383 BACs was selected and end-sequenced. The physical map consists of 154 contigs and 4,268 singletons. Forty-nine contigs are genetically anchored and ordered to chromosomes for a total span of 307.2 Mbp. The unanchored contigs (105) span 67.4 Mbp and therefore the estimated genome size of T. cacao is 374.6 Mbp. A comparative analysis with A. thaliana, V. vinifera, and P. trichocarpa suggests that comparisons of the genome assemblies of these distantly related species could provide insights into genome structure, evolutionary history, conservation of functional sites, and improvements in physical map assembly. A comparison between the two T. cacao cultivars Matina 1-6 and Criollo indicates a high degree of collinearity in their genomes, yet rearrangements were also observed. CONCLUSIONS: The results presented in this study are a stand-alone resource for functional exploitation and enhancement of Theobroma cacao but are also expected to complement and augment ongoing genome-sequencing efforts. This resource will serve as a template for refinement of the T. cacao genome through gap-filling, targeted re-sequencing, and resolution of repetitive DNA arrays.

Characteristics of patients losing vision after 2 years of monthly dosing in the phase III ranibizumab clinical trials.

PURPOSE: To investigate the cause of visual acuity (VA) loss in patients with neovascular age-related macular degeneration (AMD) receiving monthly ranibizumab injections in the pivotal ranibizumab phase III trials. DESIGN: Retrospective analysis. PARTICIPANTS: The Minimally Classic/Occult Trial of the Anti-VEGF Antibody Ranibizumab In the treatment of Neovascular AMD (MARINA) and Anti-VEGF Antibody for the Treatment of Predominantly Classic Choroidal Neovascularization in AMD (ANCHOR) trials. METHODS: Demographics and lesion characteristics at baseline and month 24 were compared in patients with ≥15 letters VA loss and patients with ≥15 letters VA gain from baseline to month 24. Additional evaluations of fundus photographs from these patients were performed to assess features of non-exudative AMD, such as geographic atrophy (GA) and retinal pigment epithelium (RPE) abnormalities. MAIN OUTCOME MEASURES: Differences in lesion characteristics between patients who lost versus gained ≥15 letters of VA from baseline to month 24. RESULTS: At month 24, 9% of ranibizumab-treated patients from MARINA and 10% of ranibizumab-treated patients from ANCHOR had lost ≥15 letters VA; 30% of ranibizumab-treated patients from MARINA and 38% of ranibizumab-treated patients from ANCHOR had gained ≥15 letters VA. Baseline characteristics associated with VA loss at month 24 included older age, better VA, and larger lesions. At month 24, an increased area of RPE abnormality was associated with VA loss in both the MARINA (P = 0.0008) and ANCHOR (P = 0.0046) trials. Increased total lesion area at month 24 was associated with VA loss in both trials. In MARINA, the increase in total lesion area was attributable to an increase in the angiographic designation of atrophic scar among VA losers (P = 0.0043), but in ANCHOR it was attributable to an increased area of choroidal neovascularization (CNV) (P = 0.039) but not an increased area of leakage (P = 0.17). Increased areas of GA, fibrosis, and hemorrhage were not associated with VA loss. CONCLUSIONS: Vision loss after 2 years of monthly ranibizumab therapy was associated with lesion characteristics commonly associated with suppressed CNV, such as pigmentary abnormalities, atrophic scar, and the absence of leakage. Future VA improvements in patients receiving ranibizumab therapy may require preservation of photoreceptor and RPE function rather than strategies that target CNV.

Public health response to striking solid waste management.

In 2009, the City of Toronto, Ontario, Canada, experienced a six-week labor disruption involving 24,000 city workers that included solid waste and public health employees. In an attempt to control illegal dumping and to manage garbage storage across the city during this period, 24 temporary garbage storage sites were established by the city (mostly in local parks) for residents to dispose of their household waste. No other municipality in North America has attempted to operate this many temporary sites for this long a period. Management and nonunion staff from Healthy Environments in Toronto Public Health performed daily inspections, responded to community questions, issued public health orders, and worked closely with Solid Waste Management and the Ministry of the Environment to actively manage the public health concerns associated with these sites. This intensive oversight mitigated public health risks to the community and facilitated an effective, safe solution to the temporary garbage storage problem.

Dosing regimen and the frequency of macular hemorrhages in neovascular age-related macular degeneration treated with ranibizumab.

PURPOSE: The purpose of this study was to investigate if monthly intravitreal ranibizumab decreases risk of macular hemorrhages in patients with choroidal neovascularization secondary to age-related macular degeneration. METHODS: Incidences of macular hemorrhages in the control and ranibizumab groups from three, multicenter, randomized, clinical trials (MARINA, ANCHOR, and PIER) were compared. Two time intervals (Months 0-3 and 5-17) were evaluated to account for transition from monthly to quarterly injections in PIER. Time interval after Month 17 was excluded because of crossover from control to active treatment in all trials. RESULTS: Months 0-3: All trials showed higher incidence rates of hemorrhages in control compared with ranibizumab groups (ANCHOR: photodynamic therapy [27.3%], 0.3 mg [8.0%], 0.5 mg [8.6%]; MARINA: sham [18.6%], 0.3 mg [8.8%], 0.5 mg [8.8%]; and PIER: sham [16.1%], 0.3 mg [3.4%], 0.5 mg [3.3%]). In ANCHOR and MARINA, data of Months 5-17 showed higher incidence rates in control compared with monthly ranibizumab groups (ANCHOR: photodynamic therapy [47.8%], 0.3 mg [12.5%], 0.5 mg [12.3%]; and MARINA: sham [38.0%], 0.3 mg [13.2%], 0.5 mg [13.0%]), but this was not seen for quarterly ranibizumab groups in PIER (sham [22.4%], 0.3 mg [23.7%], 0.5 mg [28.3%]). CONCLUSION: Treatment with monthly intravitreal ranibizumab was associated with reduced risk of new macular hemorrhages when compared with photodynamic therapy (ANCHOR) or sham (MARINA and PIER). There was no difference between PIER quarterly ranibizumab-treated and sham patients.