Silicone Sphere Implant Extrusion From Orbital Granulomatosis With Polyangiitis: A Rare Complication in the Anophthalmic Socket.

Orbital implant extrusion is a known complication following evisceration and enucleation. In this case report, we present a 45-year-old woman who presented with a left silicone implant exposure and infection 2 years following evisceration with saddle nose on examination. CT of the maxillofacial bones without contrast showed bilateral soft tissue infiltration around the superior recti muscles, as well as a nasal septum perforation from extensive sinus disease. Left orbitotomy revealed a small fibrotic mass near the orbital roof. Biopsy and serology results were consistent with granulomatosis with polyangiitis.

An intracameral approach for recalcitrant fungal keratitis.

PURPOSE: To describe 2 cases of recalcitrant fungal keratitis successfully treated with intracameral Amphotericin B. METHODS: Interventional case series. RESULTS: A 59-year-old female and a 41-year-old male each presented with fungal keratitis, caused by Bipolaris spp. and Fusarium spp. respectively. Both cases were unresponsive to topical antifungals, causing persistent discomfort and decreased vision. The two patients subsequently received a single dose of intracameral amphotericin B (ICAMB) 10mcg/0.1 mL, in addition to continued topical natamycin. Both patients had remarkable results following ICAMB, with best corrected visual acuity of 20/20 and full corneal reepithelization following treatment. CONCLUSIONS: We report 2 cases of intractable fungal keratitis that benefited from intracameral injections of amphotericin B. This route of delivery appears to be very effective because the medication is delivered directly to the deeper layers of the cornea, where fungal infections tend to reside, and where topical and systemic routes have difficulty accessing.

FEATURES AND OUTCOMES OF EYES THAT UNDERWENT SURGICAL REPAIR OF RHEGMATOGENOUS RETINAL DETACHMENTS AFTER BEING TREATED FOR ACUTE ENDOPHTHALMITIS.

PURPOSE: To evaluate the etiology, clinical course, and outcomes of eyes that suffered postendophthalmitis rhegmatogenous retinal detachments. METHODS: A retrospective, consecutive case series was conducted of patients managed at Associated Retinal Consultants P.C. from January 2013 to December 2019. Patients were identified as having had endophthalmitis by ICD-9/10 codes. Those with endophthalmitis and/or rhegmatogenous retinal detachment not managed at Associated Retinal Consultants from January were excluded. RESULTS: Charts of 413 patients were reviewed and 19 met inclusion criteria. Incidence of rhegmatogenous retinal detachment following infectious endophthalmitis was 4.6%. The most common inciting events for endophthalmitis was intravitreal injection (9 of 19) and cataract surgery (7 of 19). Fifteen of 19 patients were treated with an injection of intravitreal antibiotics and 4 underwent immediate vitrectomy with antibiotic injection. Biopsy cultures were obtained in 18 of 19 patients and yielded positive growth in 12 (66.7%). Seventeen of the 19 eyes were operable. Final retinal reattachment rate was 88.2% (15 of 17). Mean final logMAR visual acuity was 1.58 (Snellen 20/765). Factors associated with worse final visual acuity after surgical repair included preceding intravitreal injection (P = 0.001), streptococcus species (P = 0.024), presence of proliferative vitreoretinopathy (P = 0.015), and use of silicone oil during primary rhegmatogenous retinal detachment repair (P = 0.010). CONCLUSION: Rhegmatogenous retinal detachments following endophthalmitis occur infrequently. Although most eyes can be repaired surgically, visual outcomes are often poor, particularly in eyes that were infected with streptococcal species and had associated proliferative vitreoretinopathy.

Clinical Course and Characteristics of Eyes with Recurrent Episodes of Endophthalmitis.

PURPOSE: Infectious endophthalmitis is a devastating, yet rare, complication after intraocular surgery, trauma, and systemic illness. Given its rare incidence, few patients would be expected to experience more than 1 episode of infectious endophthalmitis in their lifetime. We reviewed our patients who were diagnosed with and treated for at least 2 separate episodes of endophthalmitis. DESIGN: A retrospective, consecutive case series was conducted of patients managed at Associated Retinal Consultants PC (Royal Oak, Michigan) from January 2013 through December 2019. PARTICIPANTS: Patients were identified with the diagnosis of endophthalmitis by International Classification of Diseases, Ninth and Tenth Editions, codes. METHODS: Those diagnosed and then treated either with a vitreous tap and intravitreal injection of antibiotics or with pars plana vitrectomy at least twice were included. Those treated multiple times for the same episode of endophthalmitis were excluded. MAIN OUTCOME MEASURES: Cause and risk factors for recurrent endophthalmitis. RESULTS: Charts of 535 patients were reviewed, and 12 patients met inclusion criteria. The median age at initial presentation was 72.5 years, and 33.3% were men. Eight of the 12 patients (66%) experienced recurrent endophthalmitis in the same eye, and 4 of the 12 patients (33%) experienced separate episodes in different eyes. The average time between episodes was 604 days (range, 90-2366 days). The average follow-up from the second episode was 492 days (range, 119-1185 days). The most common cause for both the first and second episodes was recent intravitreal injection (50% and 58.3%, respectively) followed by surgery associated (41.6% and 33.3%, respectively). The cause was the same for the first and second episodes of 8 patients (75%). Of the 24 recorded episodes of endophthalmitis, culture results were positive in 41.6%, with coagulase-negative Staphylococcus being the most common bacteria identified. CONCLUSIONS: Recurrent endophthalmitis is rare and seen most commonly after intravitreal injections. Most patients in this series showed culture-negative results. Each successive episode of endophthalmitis was associated with a worse final visual outcome. The cumulative number of intravitreal injections may be an independent risk factor for recurrent postinjection endophthalmitis.

Fluorescence-activated droplet sorting of lipolytic microorganisms using a compact optical system.

Lipases are ubiquitous enzymes of great physiological significance that have been used extensively in multiple industries. Environmental microorganisms are a major source for the discovery of novel lipases with high catalytic efficiency and selectivity. However, current plate-based screening of lipase-producing strains is time consuming, labour intensive and inefficient. In this study, we developed an ultra-high throughput screening pipeline for lipase-producing strains based on fluorescence-activated droplet sorting (FADS) using a compact optical system that could be easily set up in an alignment-free manner. The pipeline includes droplet generation, droplet incubation, picoinjection of the fluorescence probe, and sorting of droplets with a throughput of 2 × 106 drops per h. We applied the pipeline to screen samples collected from different locations, including sediments from a hot spring in Tibet, soils from the Zoige wetland, contaminated soils from an abandoned oilfield, and a Chinese Daqu starter. In total, we obtained 47 lipase-producing bacterial strains belonging to seven genera, including Staphylococcus, Bacillus, Enterobacter, Serratia, Prolinoborus, Acinetobacter, and Leclercia. We believe that this FADS-based pipeline could be extended to screen various enzymes from the environment, and may find wide applications in breeding of industrial microorganisms.

Automated Chemotactic Sorting and Single-cell Cultivation of Microbes using Droplet Microfluidics.

We report a microfluidic device for automated sorting and cultivation of chemotactic microbes from pure cultures or mixtures. The device consists of two parts: in the first part, a concentration gradient of the chemoeffector was built across the channel for inducing chemotaxis of motile cells; in the second part, chemotactic cells from the sample were separated, and mixed with culture media to form nanoliter droplets for encapsulation, cultivation, enumeration, and recovery of single cells. Chemotactic responses were assessed by imaging and statistical analysis of droplets based on Poisson distribution. An automated procedure was developed for rapid enumeration of droplets with cell growth, following with scale-up cultivation on agar plates. The performance of the device was evaluated by the chemotaxis assays of Escherichia coli (E. coli) RP437 and E. coli RP1616. Moreover, enrichment and isolation of non-labelled Comamonas testosteroni CNB-1 from its 1:10 mixture with E. coli RP437 was demonstrated. The enrichment factor reached 36.7 for CNB-1, based on its distinctive chemotaxis toward 4-hydroxybenzoic acid. We believe that this device can be widely used in chemotaxis studies without necessarily relying on fluorescent labelling, and isolation of functional microbial species from various environments.

High-Throughput Single-Cell Cultivation on Microfluidic Streak Plates.

This paper describes the microfluidic streak plate (MSP), a facile method for high-throughput microbial cell separation and cultivation in nanoliter sessile droplets. The MSP method builds upon the conventional streak plate technique by using microfluidic devices to generate nanoliter droplets that can be streaked manually or robotically onto petri dishes prefilled with carrier oil for cultivation of single cells. In addition, chemical gradients could be encoded in the droplet array for comprehensive dose-response analysis. The MSP method was validated by using single-cell isolation of Escherichia coli and antimicrobial susceptibility testing of Pseudomonas aeruginosa PAO1. The robustness of the MSP work flow was demonstrated by cultivating a soil community that degrades polycyclic aromatic hydrocarbons. Cultivation in droplets enabled detection of the richest species diversity with better coverage of rare species. Moreover, isolation and cultivation of bacterial strains by MSP led to the discovery of several species with high degradation efficiency, including four Mycobacterium isolates and a previously unknown fluoranthene-degrading Blastococcus species.

The intracellular Ca²âº channel MCOLN1 is required for sarcolemma repair to prevent muscular dystrophy.

The integrity of the plasma membrane is maintained through an active repair process, especially in skeletal and cardiac muscle cells, in which contraction-induced mechanical damage frequently occurs in vivo. Muscular dystrophies (MDs) are a group of muscle diseases characterized by skeletal muscle wasting and weakness. An important cause of these group of diseases is defective repair of sarcolemmal injuries, which normally requires Ca(2+) sensor proteins and Ca(2+)-dependent delivery of intracellular vesicles to the sites of injury. MCOLN1 (also known as TRPML1, ML1) is an endosomal and lysosomal Ca(2+) channel whose human mutations cause mucolipidosis IV (ML4), a neurodegenerative disease with motor disabilities. Here we report that ML1-null mice develop a primary, early-onset MD independent of neural degeneration. Although the dystrophin-glycoprotein complex and the known membrane repair proteins are expressed normally, membrane resealing was defective in ML1-null muscle fibers and also upon acute and pharmacological inhibition of ML1 channel activity or vesicular Ca(2+) release. Injury facilitated the trafficking and exocytosis of vesicles by upmodulating ML1 channel activity. In the dystrophic mdx mouse model, overexpression of ML1 decreased muscle pathology. Collectively, our data have identified an intracellular Ca(2+) channel that regulates membrane repair in skeletal muscle via Ca(2+)-dependent vesicle exocytosis.

[Micro-droplet characterization and its application for amino acid detection in droplet microfluidic system].

Recently, the droplet microfluidic system attracts interests due to its high throughput and low cost to detect and screen. The picoliter micro-droplets from droplet microfluidics are uniform with respect to the size and shape, and could be used as monodispensed micro-reactors for encapsulation and detection of single cell or its metabolites. Therefore, it is indispensable to characterize micro-droplet and its application from droplet microfluidic system. We first constructed the custom-designed droplet microfluidic system for generating micro-droplets, and then used the micro-droplets to encapsulate important amino acids such as glutamic acid, phenylalanine, tryptophan or tyrosine to test the droplets' properties, including the stability, diffusivity and bio-compatibility for investigating its application for amino acid detection and sorting. The custom-designed droplet microfluidic system could generate the uniformed micro-droplets with a controllable size between 20 to 50 microm. The micro-droplets could be stable for more than 20 h without cross-contamination or fusion each other. The throughput of detection and sorting of the system is about 600 micro-droplets per minute. This study provides a high-throughput platform for the analysis and screening of amino acid-producing microorganisms.