The Effect of Face Mask Wear on the Ocular Surface and Contact Lens Microbiome.

OBJECTIVES: As face mask wear can result in the redirection of nasal and oral exhalation toward the ocular region, this study investigated the impact of face mask wear on the conjunctiva, eyelid margin, and contact lens (CL) surface microbiome. METHODS: In this prospective, cross-over study, experienced CL wearers (N=20) were randomized to wear a face mask for 6 hr/day (minimum) for a week or no mask for a week. The conjunctiva, eyelid, and CLs were then sampled. After a 1-week washout period, participants were crossed over into the alternate treatment for 1 week and sampling was repeated. Sampling was bilateral and randomly assigned to be processed for culturing or 16S ribosomal(r) RNA gene sequencing. RESULTS: Culturing showed no effect of mask wear on the average number of bacterial colonies isolated on the conjunctiva, eyelid, or CL, but there was increased isolation of Staphylococcus capitis on CL samples with mask wear (P=0.040). Culture-independent sequencing found differences in the taxonomic complexity and bacterial composition between the three sites (P<0.001), but there was no effect of bacterial diversity within and between sites. Mask wear did not impact dry eye or CL discomfort, but increased ocular surface staining was reported (P=0.035). CONCLUSIONS: Mask wear did not substantially alter the microbiome of the conjunctiva, eyelid margin, or CL surfaces in uncompromised healthy eyes.

Detection of SARS-CoV-2 binding receptors and miscellaneous targets as well as mucosal surface area of the human lacrimal drainage system.

PURPOSE: Our aim was to evaluate a potential role for the lacrimal drainage system (LDS) as a portal of entry and conduit for SARS-CoV-2 in human infection. We also investigate the mucosal surface area. The relatively long tear contact time in a closed system raises the possibility that this pathway may contribute to the initiation of systemic infection. We looked for expression of ACE2, the main receptor for SARS-CoV-2, as well as cofactors such as TMPRSS2 and other enzymes such as cathepsinB, CD147, elastase1, furin, neuropilin1, neuropilin2, TMPRSS11D and trypsin which also play a role in SARS-CoV-2 infection, in this system. METHODS: Human tissue samples of the draining tear ducts from body donors were analyzed by RT-PCR, Western blot and immunohistochemistry. It is not known whether the respective body donors were Sars-Cov-2 positive at any time; they were negative when they entered the institute. Besides, the draining LDS of body donors were measured to determine the mucosal surface in the lacrimal system. RESULTS: The expression of the main receptor studied, ACE2, cofactors such as TMPRSS2 and other enzymes such as cathepsinB, CD147, elastase1, furin, neuropilin1, neuropilin2, TMPRSS11D and trypsin were all detected at the gene and protein level. The average mucosal surface area of the lacrimal sac and nasolacrimal duct was calculated to be 110 mm2. CONCLUSION: The results show the presence of all analyzed receptors in the efferent LDS. With an average tear passage time of 3 min and a relatively large mucosal surface area, the LDS could therefore be considered as a portal of entry and conduit for SARS-CoV-2. In addition, it represents a surface that should be taken into consideration in the administration of topically applied medication to the ocular surface.

Examining the fabric of the eye: Antoni van Leeuwenhoek, the draper and ocular microscopist.

We review Antoni van Leeuwenhoek's (1632 - 1723) microscopic studies of comparative ocular anatomy in humans, mammals, birds, and fish. His contributions in anatomical microscopy to ocular biology has been overshadowed by his prolific work and first observations of protists and bacteria, spermatozoa, red blood cells, and dental plaque. Leeuwenhoek's Delftian optical and artisanal heritage more than compensated for any lack of formal scientific training and, in keeping with his Royal Society ethos, shone light onto the "fabric of the eye" in order to better understand its function, which he had extended with his microscopes. He has provided the earliest known microscopic descriptions of the tear film, eyelids, cornea, aqueous, crystalline lens, vitreous, retina, optic nerve, and photoreceptor--as well as the first descriptions of cataract, vitreous floater,s and corneal pathology. His description of the aquatic mammal Cetacean eye foreshadowed an understanding of the optical complexities of aerial and submarine vision of whales. His observations challenged classical teaching at that time, particularly in relation to the optic nerve.

Exploring the relationship between 24-2 visual field and widefield optical coherence tomography data across healthy, glaucoma suspect and glaucoma eyes.

PURPOSE: To utilise ganglion cell-inner plexiform layer (GCIPL) measurements acquired using widefield optical coherence tomography (OCT) scans spanning 55° × 45° to explore the link between co-localised structural parameters and clinical visual field (VF) data. METHODS: Widefield OCT scans acquired from 311 healthy, 268 glaucoma suspect and 269 glaucoma eyes were segmented to generate GCIPL thickness measurements. Estimated ganglion cell (GC) counts, calculated from GCIPL measurements, were plotted against 24-2 SITA Faster visual field (VF) thresholds, and regression models were computed with data categorised by diagnosis and VF status. Classification of locations as VF defective or non-defective using GCIPL parameters computed across eccentricity- and hemifield-dependent clusters was assessed by analysing areas under receiver operating characteristic curves (AUROCCs). Sensitivities and specificities were calculated per diagnostic category. RESULTS: Segmented linear regression models between GC counts and VF thresholds demonstrated higher variability in VF defective locations relative to non-defective locations (mean absolute error 6.10-9.93 dB and 1.43-1.91 dB, respectively). AUROCCs from cluster-wide GCIPL parameters were similar across methods centrally (p = 0.06-0.84) but significantly greater peripherally, especially when considering classification of more central locations (p < 0.0001). Across diagnoses, cluster-wide GCIPL parameters demonstrated variable sensitivities and specificities (0.36-0.93 and 0.65-0.98, respectively), with the highest specificities observed across healthy eyes (0.73-0.98). CONCLUSIONS: Quantitative prediction of VF thresholds from widefield OCT is affected by high variability at VF defective locations. Prediction of VF status based on cluster-wide GCIPL parameters from widefield OCT could become useful to aid clinical decision-making in appropriately targeting VF assessments.

High-Resolution Ocular Surface Imaging: Real-Time Visualization of Tear Film Dysfunction.

PURPOSE: Recent advancements in infrared sensing technology have made it possible to visualize tear film dynamics in real time, enabling evaluation of tear film quality during blinking. A retrospective clinical evaluation was conducted to explore this by grading videos of the tear film and comparing grading data with dry eye diagnostic results using the OCULUS keratograph (K5M). METHODS: Videos were used to grade patients' tear film perturbations as compared with healthy control subjects. The grading was then correlated with the ocular surface disease index (OSDI) scores, tear film breakup time (TFBUT), tear meniscus height (TMH), corneal staining, redness, and meibography data. RESULTS: Infrared imaging of the ocular surface revealed instantaneous and recurring dynamic characteristics of the tear film, allowing for the differentiation between normal and abnormal tear films. Abnormal features included a complete absence of a spreading tear film, hindered spreading of the tear film after blinking, areas of tear film instability, or a combination of the latter 2. Some of these features show a resemblance to the tear film appearance after fluorescein staining. The grading of these features correlated with TFBUT and, to a lesser extent, with TMH but did not show significant correlation with any other diagnostic data from the K5M. Furthermore, the speed of tear film spreading after blinking showed a positive correlation with TMH. CONCLUSIONS: Direct visualization of the tear film across the entire palpebral aperture using infrared sensing offers a noninvasive, reproducible, and rapid method for assessing the health and quality of the tear film.

Recovery From Homonymous Hemianopia in Hyperglycemia-Induced Occipital Lobe Electroclinical Seizures Following Glycemic Control.

BACKGROUND: Visual changes due to hyperglycemia in diabetes are not uncommon. While blurred vision is a well-established sequela of chronic hyperglycemia, homonymous hemianopia with or without electroclinical seizures is much rarer and can be mistaken for migraine, temporal arteritis, or ischemia of the central nervous system. METHODS: This article analyzed case studies for 3 patients (67M, 68M, 52F) presenting with complex visual phenomena, from 3 to 42 days duration, including pathogenesis, clinical findings, management, and follow-up. RESULTS: Examinations demonstrated dense left homonymous hemianopias in 2 patients and a left inferior homonymous quadrantanopia in one, with no other abnormalities. Patients described vivid, nonstereotyped intermittent hallucinations in the affected fields. Blood glucose levels ranged from 13.5 to 35.0 mmol/L (243-630 mg/dL) without ketosis and HbA1c from 14.6% to 16.8%. Computed tomography of the brain showed no acute intracranial pathology. MRI of the brain either detected no abnormalities or demonstrated changes consistent with seizure activity. Electroencephalogram (EEG) demonstrated seizures over the right occipital region in each patient. EEG seizures coincided with patients' hallucinations, while they remained otherwise conscious. Oral hypoglycemic and antiepileptic medications were commenced with rapid and complete reversal of the seizures and visual field deficits, confirmed by repeat Automated 30-2 and MRI. CONCLUSIONS: Hyperglycemia-induced occipital lobe seizures with visual hallucinations and interictal homonymous visual field defects represent a rare but clinically important diagnosis. This article highlights the importance of prompt recognition and treatment to facilitate recovery.

Impact of Various Concentrations of Low-Dose Atropine on Pupillary Diameter and Accommodative Amplitude in Children with Myopia.

Purpose: To assess over 2 weeks, the effect of 3 different low concentrations of atropine on pupillary diameter and accommodative amplitude in children with myopia. Methods: Fifty-eight children with myopia [spherical equivalent (SE) of -0.50 diopters (D) or worse, astigmatism of less than or equal to 2.00 D] were randomly allocated to 3 groups receiving 0.01%, 0.02%, or 0.03% atropine eye drops, once nightly for 2 weeks. The primary outcome was the change from baseline in pupillary diameter and accommodative amplitude with each of the concentrations. Results: Fifty-seven participants (114 eyes), aged between 6 and 12 years, completed the 2-week trial (mean age 9.3 ± 1.7 years and mean SE -3.53 ± 1.79 D). After 2 weeks of use, all the 3 concentrations were found to have a statistically significant effect on both the pupillary diameter and accommodative amplitude. Accommodative amplitude reduced by an average of 5.23 D, 9.28 D, and 9.32 D, and photopic pupil size increased by an average of 0.95 ± 1.05 mm, 1.65 ± 0.93 mm, and 2.16 ± 0.88 mm with 0.01%, 0.02%, and 0.03%, respectively. Of the eyes, a total of 5.3% and 5.9% of the eyes on 0.02% and 0.03% atropine had a mean residual accommodative amplitude of <5 D. The percentage of eyes having a pupillary dilation >3 mm were 4.8%, 10.5%, and 23.5% for 0.01%, 0.02%, and 0.03% atropine, respectively. Conclusions: Low-dose atropine had an effect on pupillary diameter and accommodative amplitude. With the highest concentration assessed, that is, 0.03% nearly 1 of 4 eyes had pupillary dilation of >3 mm. Clinical Trial Registration number: NCT03699423.

A Systematic Review of Ophthalmology Education in Medical Schools: The Global Decline.

TOPIC: This systematic review examined geographical and temporal trends in medical school ophthalmology education in relationship to course and student outcomes. CLINICAL RELEVANCE: Evidence suggesting a decline in ophthalmology teaching in medical schools is increasing, raising concern for the adequacy of eye knowledge across the rest of the medical profession. METHODS: Systematic review of Embase and SCOPUS, with inclusion of studies containing data on medical school ophthalmic course length; 1 or more outcome measures on student ophthalmology knowledge, skills, self-evaluation of knowledge or skills, or student course appraisal; or both. The systematic review was registered prospectively on the International Prospective Register of Systematic Reviews (identifier, CRD42022323865). Results were aggregated with outcome subgroup analysis and description in relationship to geographical and temporal trends. Descriptive statistics, including nonparametric correlations, were used to analyze data and trends. RESULTS: Systematic review yielded 4596 publication titles, of which 52 were included in the analysis, with data from 19 countries. Average course length ranged from 12.5 to 208.7 hours, with significant continental disparity among mean course lengths. Africa reported the longest average course length at 103.3 hours, and North America reported the shortest at 36.4 hours. On average, course lengths have been declining over the last 2 decades, from an average overall course length of 92.9 hours in the 2000s to 52.9 hours in the 2020s. Mean student self-evaluation of skills was 51.3%, and mean student self-evaluation of knowledge was 55.4%. Objective mean assessment mark of skills was 57.5% and that of knowledge was 71.7%, compared with an average pass mark of 66.7%. On average, 26.4% of students felt confident in their ophthalmology knowledge and 34.5% felt confident in their skills. DISCUSSION: Most evidence describes declining length of courses devoted to ophthalmology in the last 20 years, significant student dissatisfaction with courses and content, and suboptimal knowledge and confidence. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Aqueous Humor Circulation in the Era of Minimally Invasive Surgery for Glaucoma.

Glaucoma surgery with implantation of aqueous humor draining microstents may compromise long-term corneal health by disrupting aqueous humor circulation. The effect of stent numbers on this circulation was interrogated to determine the number of stents associated with minimal circulation disruption. An in vitro anterior eye model perfusion system was constructed with multiple exit ports. A 3-D model of the anterior eye was imported into ABAQUS CFD, analyzes were carried out for unsteady laminar flow and solved using Navier-Stokes equations. DT Vision Foundry was used to analyze velocity contour plot images. The field variable results output for the CFD model were fluid wall shear, fluid pressure and fluid velocity. In vitro, "aqueous" fluid flow is high through a single stent and "aqueous" stagnation is greatest in the quadrants 180° away. Increasing stent port numbers, results in an exponential decrease in the stagnant flow locations. High wall shear stress was seen in the single stent model and is markedly reduced after a second and subsequent stents are introduced. We identify two factors potentially contributing to corneal compromise post glaucoma drainage surgery: aqueous humor stagnation, remote to the stent site and higher exit flows imparting increased stent exit shear stress (particularly with a single stent). With 4 stents, there is minimal disruption of anterior chamber circulation (mimicking physiological conditions). Furthermore we propose that aqueous humor circulation disruption via the usual single-exit port approach disrupts aqueous humor circulation with long-term consequences for corneal health.

Ocular microbiome changes in dry eye disease and meibomian gland dysfunction.

The most common and chronic ocular problem of aging is dry eye disease (DED) and the associated condition of meibomian gland dysfunction (MGD). The resident ocular surface bacteria may have a role in maintaining homeostasis and perturbation may contribute to disease development. The aim of this study was to compare the microbiomes of the conjunctiva and eyelid margin in humans with mild and moderate DED and controls using 16 S rRNA gene sequencing. The conjunctiva and lid margin of three cohorts (N = 60; MGD, MGD with lacrimal dysfunction [MGD + LD] and controls) were swabbed bilaterally three times over three months. Microbial communities were analysed by extracting DNA and sequencing the V3-V4 region of the 16 S ribosomal RNA gene using the Illumina MiSeq platform. Sequences were quality filtered, clustered into amplicon sequence variants (ASVs) using UNOISE algorithm and taxonomically classified using a Bayesian Last Common Ancestor (BCLA) algorithm against the GTDB 2207 database. The overall microbial communities of the MGD, MGD + LD and control groups were significantly different from each other (P = 0.001). The MGD and MGD + LD dry eye groups showed greater variability between individuals compared to the control (PERMDISP, P < 0.01). There was decreased richness and diversity in females compared to males for the conjunctiva (P < 0.04) and eyelid margin (P < 0.018). The conjunctiva in the MGD + LD group had more abundant Pseudomonas azotoformans, P. oleovorans and Caballeronia zhejiangensis compared to MGD and control (P < 0.05), while the MGD group had more abundant Corynebacterium macginleyi and C. kroppenstedtii compared to control (P < 0.05). The lid margin in MGD was more abundant in C. macginleyi, C. accolens, and C. simulans compared to the MGD + LD and control (P < 0.05). There were differences in the overall microbial community composition and certain taxa, including increased levels of lipophilic bacteria, on the conjunctiva and eyelid margin in mild to moderate DED/MGD compared to controls. DED/MGD was also associated with a reduced bacterial richness and diversity in females.

Modified Limbal-Conjunctival Autograft Surgical Technique: Long-Term Results of Recurrence and Complications.

PURPOSE: The aim of this study was to report the recurrence and complication rates of a modified limbal-conjunctival autograft surgical technique for pterygium excision. METHODS: This was a retrospective, single-surgeon, single-operating environment, consecutive case series of 176 eyes in 163 patients with a biopsy-proven diagnosis of pterygium. All patients underwent excision using a 23-gauge needle to "behead" the pterygium head, followed by a limbal-conjunctival autograft including ∼50% of the palisades of Vogt. Outcomes measured included recurrence, defined as any conjunctival fibrovascular growth, and complication rates. Correlations between preoperative patient characteristics, pterygium morphology, and intraoperative factors (width of corneal extension, conjunctival defect, and graft) with postoperative recurrence were examined using logistic regression models. RESULTS: The median age was 59.5 years and 122 eyes (69.3%) had primary pterygium (type I: 17%, II: 37.5%, and III: 45.5%). Kaplan-Meier analysis demonstrated the median pterygium-free follow-up period to be 723 days (range 46-7230 days). Recurrence was observed in 3 eyes of 2 patients (1.7%). No postoperative graft-related complications were observed. Postoperative symptomatology was transient. Age demonstrated a negative correlation with recurrence (odds ratio 0.888, 95% CI, 0.789-0.998, P = 0.046). However, no other correlations with preoperative or intraoperative factors, including whether pterygium was primary or recurrent, were identified (all P > 0.05). CONCLUSIONS: This modified limbal-conjunctival autograft technique represents an effective alternative that offers a very low recurrence rate and avoids extensive dissection or antimetabolites, with minimal complications and transient postoperative symptomatology, over a long-term follow-up period. This technique is relatively simple and successful for both primary and recurrent pterygia. Future comparative studies with other surgical techniques may determine which are superior.

Case Series of Rare Fungal Keratitides: Experiences from a Quaternary Eye Hospital in Sydney, Australia.

The present article reports on the management of six different and rare cases of fungal keratitides, two of which have never been documented in previous literature. This is a case series of six patients with rare fungal keratitides managed at a quaternary eye referral unit, Sydney Eye Hospital, Australia over a period of 7 months (May to December, 2022). The order of occurrence of fungi isolated was Scedosporium apiospermum, Lomenstospora prolificans, Cladosporium spp., Paecilomyces, Syncephalastrum racemosum and Quambalaria spp. A combination of medical and surgical interventions was employed, including topical and systemic anti-fungal therapy, with one requiring therapeutic penetrating keratoplasty and another eventuating in evisceration. Two patients were successfully treated with corneal debridement and two others required pars plana vitrectomy with anterior chamber washout. It is important to remain vigilant with monitoring patient symptoms and correlating with clinical signs to guide antifungal therapy even in the context of confirmed culture and sensitivity results.

Pseudo-hallucinations are for real in some patients with a bipolar disorder.

OBJECTIVE: To provide exemplars of pseudo-hallucinations experienced by six individuals with bipolar disorder, suggest that they may be preferentially overrepresented in those with that condition and argue that their presence does not indicate that the individual is necessarily psychotic. METHOD: Historical definitions of pseudo-hallucinations are overviewed and six profiles are presented considering manifestations in those with a bipolar disorder, their sequencing and response to medication. RESULTS: The six patients initially judged such experiences as true hallucinations and, only over time, did they recognize their falsity. Clinical observation suggests that pseudo-hallucinations may be more common in those with a bipolar disorder and that they can precede, cooccur or follow disorder onset, can manifest during any phase of the condition and that they may or may not respond to medication prescribed for the bipolar disorder. CONCLUSIONS: The exemplars of pseudo-hallucinations overviewed here indicate their richness of expression, their variable time course with illness stages and their variable response to medications prescribed for the bipolar condition.

Zonular and capsular bag disorders: a hypothetical perspective based on recent pathophysiological insights.

The purpose of this article was to look at the pathophysiology behind and devise a classification system for the causes of zonular apparatus-capsular bag (ZACB) insufficiency. Also discussed is dystrophic bag syndrome, including clinical cases and addressing where it lies on the ZACB spectrum. There has been interest in the emergence of in-the-bag intraocular lens (IOL) subluxation, the prevalence of which is increasing. There has also been a recent report of dead bag syndrome, which the authors believe is part of the same disease spectrum. The authors put these phenomena into perspective and provide a classification system based on the possible causes of what they have termed ZACB insufficiency. The basic aspects of capsular bag-IOL ocular pathophysiology are summarized with a focus on functional aspects and the consequences for IOL fastening. Within this framework, dystrophic bag syndrome is a form of primary capsular ZACB insufficiency. The contribution of factors such as intraocular drugs may suggest a reconsideration of agents used and their mode of application.

Conjunctival ultraviolet autofluorescence area decreases with age and sunglasses use.

BACKGROUND: Conjunctival ultraviolet autofluorescence (CUVAF) is a method of detecting conjunctival damage related to ultraviolet radiation exposure. In cross-sectional studies, CUVAF area is positively associated with self-reported time spent outdoors and pterygium and negatively associated with myopia; however, longitudinal studies are scarce. AIMS: To use a novel deep learning-based tool to assess 8-year change in CUVAF area in young adults, investigate factors associated with this change and identify the number of new onset pterygia. METHODS: A deep learning-based CUVAF tool was developed to measure CUVAF area. CUVAF area and pterygium status were assessed at three study visits: baseline (participants were approximately 20 years old) and at 7-year and 8-year follow-ups. Participants self-reported sun protection behaviours and ocular history. RESULTS: CUVAF data were available for 1497 participants from at least one study visit; 633 (43%) participants had complete CUVAF data. Mean CUVAF areas at baseline and the 7-year and 8-year follow-ups were 48.4, 39.3 and 37.7 mm2, respectively. There was a decrease in mean CUVAF area over time (change in total CUVAF area=-0.96 mm2 per year (95% CI: -1.07 to -0.86)). For participants who wore sunglasses ≥1/2 of the time, CUVAF area decreased by an additional -0.42 mm2 per year (95% CI: -0.72 to -0.12) on average. Fourteen (1.5%) participants developed a pterygium. CONCLUSIONS: In this young adult cohort, CUVAF area declined over an 8-year period. Wearing sunglasses was associated with a faster reduction in CUVAF area. Deep learning-based models can assist in accurate and efficient measurement of CUVAF area.

Prevalence of cataract among Australian commercial airline pilots.

Because little is known about cataract in pilots, we estimated prevalence by anonymously ascertaining all commercial airline pilots diagnosed with cataract 2011-2016 using the electronic Medical Records System of the Australian Civil Aviation Safety Authority. Of 14,163 Australian male commercial pilots licensed in 2011, 1286 aged ≥60 had biennial eye examinations showing a cataract prevalence of 11.6%. Among 12,877 pilots aged <60, based on compulsory eye examinations only when first licensed, prevalence was 0.5%. There was no significant difference by ambient ultraviolet (UV) radiation levels in state of residence though lowest prevalence was seen in the low-UV state of Victoria. Most cataract in pilots ≥60 years was bilateral and of mild severity, while cataract in pilots <60 were more likely to be unilateral and of greater severity.

Electron Microscopy and Elemental Analysis of Defective Drysdale Nucleus Manipulator Instruments Implicated in Surgical Complications.

PURPOSE: To analyze microtopography of 5 reusable Drysdale nucleus manipulator (DNM) paddled tips for sharp defects and evaluate their elemental composition to determine probable source, investigating 2 instruments (DNM 1 and 4) implicated in causing posterior capsule rupture (PCR) and 3 instruments with sharp edges identified by finger-tip interrogation intraoperatively. DESIGN: Experimental laboratory investigation. METHODS: DNM paddled tips were analyzed using scanning electron microscopy (SEM) to evaluate for sharp surface defects (number, dimensions), and subsequently energy dispersive x-ray spectroscopy (EDS) performed on sharp defects to determine their elemental composition. RESULTS: All reused DNMs analyzed (5 of 5) had significant structural defects on SEM analysis including sharp burrs, cavities and indentations, surface debris or residues, and roughening, compared to the new instrument (DNM 3, control) which had no defects. DNM 1 had 2 sharp defects, a larger 14 × 76-µm one and a craterlike 167 × 220-µm defect containing debris. EDS found that DNM 2 had 3 of 4 burrs composed mainly of carbon, the fourth of mixed composition (calcium, sulfur, oxygen); DNM 4 had 1 small burr, EDS significant for carbon; DNM 5 had 3 prominent burrs, the largest of 20 × 28 µm, 2 composed of aluminum, and some carbon residue. DNM 6 had 1 burr composed of aluminum and 3 prominent cavity defects, the largest covering 781 µm2. CONCLUSION: Reusable DNMs are widely used in cataract surgery. Sharp carbon- or aluminum-containing burrs were detected on all reused instruments analyzed together with 1 burr of mixed composition, originating from (1) organic residues, (2) instrument fragments, or (3) salt and contaminant deposits. Sharp defects may contribute to capsular damage including PCR, and residues may pose other safety concerns. Therefore, we support development of a quality, reliable single-use alternative instrument and further encourage careful inspection of all reusable instruments principally by finger-tip interrogation for sharp edges preuse.