FastRNA: An efficient solution for PCA of single-cell RNA-sequencing data based on a batch-accounting count model.

Almost always, the analysis of single-cell RNA-sequencing (scRNA-seq) data begins with the generation of the low dimensional embedding of the data by principal-component analysis (PCA). Because scRNA-seq data are count data, log transformation is routinely applied to correct skewness prior to PCA, which is often argued to have added bias to data. Alternatively, studies have proposed methods that directly assume a count model and use approximately normally distributed count residuals for PCA. Despite their theoretical advantage of directly modeling count data, these methods are extremely slow for large datasets. In fact, when the data size grows, even the standard log normalization becomes inefficient. Here, we present FastRNA, a highly efficient solution for PCA of scRNA-seq data based on a count model accounting for both batches and cell size factors. Although we assume the same general count model as previous methods, our method uses two orders of magnitude less time and memory than the other count-based methods and an order of magnitude less time and memory than the standard log normalization. This achievement results from our unique algebraic optimization that completely avoids the formation of the large dense residual matrix in memory. In addition, our method enjoys a benefit that the batch effects are eliminated from data prior to PCA. Generating a batch-accounted PC of an atlas-scale dataset with 2 million cells takes less than a minute and 1 GB memory with our method.

Pseudobulk with proper offsets has the same statistical properties as generalized linear mixed models in single-cell case-control studies.

MOTIVATION: Generalized linear mixed models (GLMMs), such as the negative-binomial or Poisson linear mixed model, are widely applied to single-cell RNA sequencing data to compare transcript expression between different conditions determined at the subject level. However, the model is computationally intensive, and its relative statistical performance to pseudobulk approaches is poorly understood. RESULTS: We propose offset-pseudobulk as a lightweight alternative to GLMMs. We prove that a count-based pseudobulk equipped with a proper offset variable has the same statistical properties as GLMMs in terms of both point estimates and standard errors. We confirm our findings using simulations based on real data. Offset-pseudobulk is substantially faster (>×10) and numerically more stable than GLMMs. AVAILABILITY AND IMPLEMENTATION: Offset pseudobulk can be easily implemented in any generalized linear model software by tweaking a few options. The codes can be found at https://github.com/hanbin973/pseudobulk_is_mm.

MarcoPolo: a method to discover differentially expressed genes in single-cell RNA-seq data without depending on prior clustering.

The standard analysis pipeline for single-cell RNA-seq data consists of sequential steps initiated by clustering the cells. An innate limitation of this pipeline is that an imperfect clustering result can irreversibly affect the succeeding steps. For example, there can be cell types not well distinguished by clustering because they largely share the global structure, such as the anterior primitive streak and mid primitive streak cells. If one searches differentially expressed genes (DEGs) solely based on clustering, marker genes for distinguishing these types will be missed. Moreover, clustering depends on many parameters and can often be subjective to manual decisions. To overcome these limitations, we propose MarcoPolo, a method that identifies informative DEGs independently of prior clustering. MarcoPolo sorts out genes by evaluating if the distributions are bimodal, if similar expression patterns are observed in other genes, and if the expressing cells are proximal in a low-dimensional space. Using real datasets with FACS-purified cell labels, we demonstrate that MarcoPolo recovers marker genes better than competing methods. Notably, MarcoPolo finds key genes that can distinguish cell types that are not distinguishable by the standard clustering. MarcoPolo is built in a convenient software package that provides analysis results in an HTML file.

Quantitative assessments of pupillary light reflexes in hospital-onset unresponsiveness.

BACKGROUND: Patients who develop hospital-onset unresponsiveness should be promptly managed in order to avoid clinical deterioration. Pupillary examination through pupillary light reflex is the gold standard method in the initial evaluation of unresponsive patients. However, the current method of shining light and subjective description often shows poor reliability. The objective of this study is to explore whether a quantitative measurement of pupillary light reflexes is useful in detecting brain herniation syndrome and predicting neurological outcomes in patients who developed hospital-onset unresponsiveness after admission for non-neurological reasons. METHODS: This was a registry-based observational study on patients who activated the neurological rapid response team at Asan Medical Center (Seoul, Korea). Hospital-onset unresponsiveness was defined as a newly developed unresponsive state as assessed by the ACDU (Alert, Confused, Drowsy, and Unresponsive) scale during the hospital stay. Demographics, comorbidities, pupillometry parameters including Neurological Pupil index, brain herniation syndrome, in-hospital mortality, and modified Rankin Scale at 3-months were analyzed. RESULTS: In 214 consecutive patients with hospital-onset unresponsiveness, 37 (17%) had brain herniation syndrome. The optimal cut-off value of Neurological Pupil index for detecting brain herniation syndrome was < 1.6 (specificity, 91% [95% confidence interval (CI) = 86-95]; sensitivity, 49% [95% CI = 32-66]). The in-hospital mortality rate was 28% (59/214); the Neurological Pupil index was negatively associated with in-hospital mortality after adjustments for the presence of brain herniation syndrome (adjusted odds ratio = 0.77, 95% CI = 0.62-0.96). Poor neurological outcomes (modified Rankin Scale ≥4) at 3 months was observed in 76% (152/201) of the patients; the Neurological Pupil index was negatively associated with poor neurological outcomes after adjustments for clinical variables (adjusted odds ratio = 0.67, 95% CI = 0.49-0.90). CONCLUSIONS: Quantitative measurements of pupillary light reflexes may be useful for early detection of potentially life-threatening neurological conditions in patients with hospital-onset unresponsiveness.

Machine Learning Approach to Identify Stroke Within 4.5 Hours.

Background and Purpose- We aimed to investigate the ability of machine learning (ML) techniques analyzing diffusion-weighted imaging (DWI) and fluid-attenuated inversion recovery (FLAIR) magnetic resonance imaging to identify patients within the recommended time window for thrombolysis. Methods- We analyzed DWI and FLAIR images of consecutive patients with acute ischemic stroke within 24 hours of clear symptom onset by applying automatic image processing approaches. These processes included infarct segmentation, DWI, and FLAIR imaging registration and image feature extraction. A total of 89 vector features from each image sequence were captured and used in the ML. Three ML models were developed to estimate stroke onset time for binary classification (≤4.5 hours): logistic regression, support vector machine, and random forest. To evaluate the performance of ML models, the sensitivity and specificity for identifying patients within 4.5 hours were compared with the sensitivity and specificity of human readings of DWI-FLAIR mismatch. Results- Data from a total of 355 patients were analyzed. DWI-FLAIR mismatch from human readings identified patients within 4.5 hours of symptom onset with 48.5% sensitivity and 91.3% specificity. ML algorithms had significantly greater sensitivities than human readers (75.8% for logistic regression, P=0.020; 72.7% for support vector machine, P=0.033; 75.8% for random forest, P=0.013) in detecting patients within 4.5 hours, but their specificities were comparable (82.6% for logistic regression, P=0.157; 82.6% for support vector machine, P=0.157; 82.6% for random forest, P=0.157). Conclusions- ML algorithms using multiple magnetic resonance imaging features were feasible even more sensitive than human readings in identifying patients with stroke within the time window for acute thrombolysis.

Are Genetic Variants Associated with the Location of Cerebral Arterial Lesions in Stroke Patients?

BACKGROUND: Genetic variants may play a role in determining the location of cerebral atherosclerosis. We aimed to investigate the association between RNF213, MMP2, and genetic polymorphisms linked to vascular tortuosity with the location of cerebral arterial atherosclerosis. METHODS: A prospective case-control study was conducted on patients with ischemic stroke and age- and sex-matched stroke-free controls. The stroke patients were categorized into those with intracranial artery atherosclerosis (ICAS), extracranial artery atherosclerosis (ECAS), and small vessel occlusion (SVO). Six single nucleotide polymorphisms (SNPs) including rs2118181 (FBN1), rs2179357 (SLC2A10), rs1036095 (TGFBR2), rs243865 (MMP2), rs1800470 (TGFB1), and rs112735431 (RNF213) were analyzed with the TaqMan Genotyping Assay, and the distribution of genotypes across groups was compared. RESULTS: None of the 6 SNPs were associated with stroke on comparing the 449 stroke patients (71 with ECAS, 169 with ICAS, and 209 with SVO) to the 447 controls. In the subgroup analysis, the adjusted odds ratios (aORs) for age and sex indicated a significant association between rs112735431 and ICAS in the allele comparison analysis and in the additive and dominant model analyses. rs112735431 was associated with anterior circulation involvement and increased burden of cerebral atherosclerosis. rs2179357 was significantly associated with ICAS in the recessive model analysis, and rs1800470 was significantly associated with ECAS in the recessive model analysis when compared to controls. CONCLUSION: rs112735431 was associated with ICAS and increased atherosclerosis burden in Korean stroke patients. Further studies are needed to elucidate the role of rs112735431 and to confirm the association of rs2179357 and rs1800470 with cerebral atherosclerosis.

New Onset Alacrima as a Presenting Feature of a Skull Base Chondrosarcoma.

Acquired unilateral alacrima as a presenting sign of an intracranial tumor is exceptionally rare, and only described once previously in a case of nasopharyngeal carcinoma. The authors present a 32-year-old female patient who presents with a year-long history of alacrima and arhinorrhea. She was subsequently diagnosed with a petroclival chondrosarcoma extending into Meckel's cave and the cavernous sinus and underwent surgical debulking. To the authors' knowledge, this is the first reported case of acquired unilateral alacrima as a presenting feature of a skull base chondrosarcoma. This case serves to remind general ophthalmologists and oculoplastic surgeons alike that acquired alacrima may be the presenting feature of serious intracranial disease.

CT and radiographic evaluation of bronchial collapsibility at forced expiration in asymptomatic brachycephalic dogs.

Bronchial collapse due to bronchomalacia is an important cause of chronic coughing in dogs. Radiographic and CT evidence of bronchial collapse has previously been reported in healthy Beagle dogs under forced expiration. However, published studies in brachycephalic dog breeds that are prone to bronchial collapse are currently lacking. In the present prospective analytical experimental study, CT and radiography were used to measure the bronchial diameter and collapsibility of each pulmonary bronchus during end-expiratory, 5 mL/kg forced-expiratory, and 10 mL/kg forced-expiratory phases in 17 asymptomatic brachycephalic dogs and six healthy Beagle dogs. Bronchial collapsibility was significantly greater during forced expiration, than that at the end of expiration in both groups (P < .001). Bronchial collapsibility measurements of the left lung lobes and the right cranial, middle, and accessory lobes were significantly higher in asymptomatic brachycephalic dogs than those in healthy Beagle dogs, during all expiratory phases (P < .05). The higher bronchial collapsibility of brachycephalic dogs was also supported using CT multiplanar reconstruction images and radiography. In conclusion, radiographic and CT measures of bronchial collapsibility in asymptomatic brachycephalic dogs are higher than measures in healthy Beagle dogs. Therefore, measures of bronchial collapse in brachycephalic dogs should not be evaluated using the same baseline measures as those used for healthy Beagle dogs.

Investigation of Postoperative Complications of Intrabony Cystic Lesions in the Oral and Maxillofacial Region.

PURPOSE: The purpose of this study was to identify factors that can complicate the surgical removal of intrabony cysts and any relevant correlations between them. PATIENTS AND METHODS: The medical records of 249 patients who underwent surgical removal of intrabony cysts were retrospectively reviewed. Outcome variables were postoperative complications, infection, and recurrence. Predictor variables were patient age, gender, comorbidities, anatomic location, pathologic diagnosis, preoperative infection, previous marsupialization, and bone graft methods. Logistic regression analysis was performed to identify risk factors of postoperative infection and recurrence. RESULTS: The cystic lesion was smallest in patients who did not receive a bone graft and increased steadily in those who received a xenogeneic bone graft and an autogenous bone graft, in that order. Paresthesia occurred after enucleation of the cystic lesion in 38 cases. Pathologic fractures were observed in 4 cases. There were 59 postoperative infections. The postoperative infection rate was as high as 63.6% in patients who underwent autogenous bone grafting. In contrast, infection rates were as low as 26.8 and 19.5% in those who underwent xenogeneic bone grafting and no bone grafting, respectively. Location of the cystic lesion in the maxilla or mandible affected the infection rate. When cysts were located in the anterior mandible, no postoperative infection occurred. In contrast, the infection rate was highest for cysts in the mandibular ramus, followed by those in the posterior mandible. Cystic lesion recurrence was observed in 7 cases: 5 cases of odontogenic keratocysts, 1 case of periapical cyst, and 1 case of dentigerous cyst. These findings suggest that cyst pathologic identity affects the recurrence rate. Cyst size was meaningfully correlated with recurrence rate. CONCLUSIONS: These results suggest that autogenous bone grafts increase the risk of postoperative infection compared with absence of a bone graft.

Experimental and Computational Study of the ( Z)-Selective Formation of Trisubstituted Olefins and Benzo-Fused Oxacycles from the Ruthenium-Catalyzed Dehydrative C-H Coupling of Phenols with Ketones.

The cationic Ru-H complex was found to be an effective catalyst for the dehydrative C-H coupling of phenols with ketones to form the trisubstituted olefin products. The coupling of phenol with linear ketones led to highly stereoselective formation of the ( Z)-olefin products. The dehydrative coupling of phenol with enones and diones efficiently formed the benzopyrene and related oxacyclic derivatives. The reaction of 3,5-dimethoxyphenol with cyclohexanone-2,2,6,6- d4 showed a significant H/D exchange to both vinyl and α-CH2 positions on the olefin product (72-75% D). A significant carbon isotope effect was observed on the ortho-arene carbon of the olefin product. The free energies of intermediate species for the entire catalytic cycle were successfully computed by using the DFT method. The DFT study revealed that the E/ Z stereoselectivity is a result of the energy difference in the insertion step of ortho-metalated phenol to an enol form of the ketone substrate (ΔΔ E = 9.6 kcal/mol). The coupling method provides a direct catalytic C-H olefination method for ketones to form trisubstituted olefins without employing any reactive reagents or forming any wasteful byproducts.

Synthesis of Symmetric and Unsymmetric Secondary Amines from the Ligand-Promoted Ruthenium-Catalyzed Deaminative Coupling Reaction of Primary Amines.

The catalytic system generated in situ from the tetranuclear Ru-H complex with a catechol ligand (1/L1) was found to be effective for the direct deaminative coupling of two primary amines to form secondary amines. The catalyst 1/L1 was highly chemoselective for promoting the coupling of two different primary amines to afford unsymmetric secondary amines. The analogous coupling of aniline with primary amines formed aryl-substituted secondary amines. The treatment of aniline- d7 with 4-methoxybenzylamine led to the coupling product with significant deuterium incorporation on CH2 (18% D). The most pronounced carbon isotope effect was observed on the α-carbon of the product isolated from the coupling reaction of 4-methoxybenzylamine (C(1) = 1.015(2)). A Hammett plot was constructed from measuring the rates of the coupling reaction of 4-methoxyaniline with a series of para-substituted benzylamines 4-X-C6H4CH2NH2 (X = OMe, Me, H, F, CF3) (ρ = -0.79 ± 0.1). A plausible mechanistic scheme has been proposed for the coupling reaction on the basis of these results. The catalytic coupling method provides an operationally simple and chemoselective synthesis of secondary amine products without using any reactive reagents or forming wasteful byproducts.

Computed tomographic and radiographic bronchial collapse may be a normal characteristic of forced expiration in dogs.

Tracheobronchomalacia has been diagnosed using radiography or bronchoscopy to confirm bronchial changes in luminal diameter during the respiratory cycle. However, studies in healthy humans suggest that some degree of bronchial collapse may be observed during the normal respiratory cycle. In this analytical study, the luminal diameter of the bronchus to each of the six pulmonary lobes and the mean percentage of expiratory collapse from end inspiratory, end expiratory, and two forced expiratory phases (10 and 15 ml/kg) were determined via computed tomography (CT) and radiography in 22 healthy Beagle dogs. The bronchial collapsibility was significantly greater during the forced expiration than the end expiration (P < 0.001); the same results were observed in dorsal and sagittal CT images and radiographs (P < 0.001). Median collapsibility values associated with 15 ml/kg forced expiratory collapse determined via cross-sectional CT images were measured as 16.6-45.5% and differed according to the pulmonary lobe. Median collapsibilities on radiography with 15 ml/kg forced expiration were 57.8% and 62.1% in the right cranial lobe and right caudal lobe, respectively. In conclusion, bronchial diameter may change during the respiratory cycle, and some degree of reduction in bronchial diameter may be an incidental finding in healthy dogs. More rigorous criteria are needed with regards to bronchial collapsibility during normal respiration for the diagnosis of bronchomalacia in order to avoid false-positive diagnoses.

Synthesis of 2-Acylphenol and Flavene Derivatives from the Ruthenium-Catalyzed Oxidative C-H Acylation of Phenols with Aldehydes.

The cationic ruthenium hydride complex [(C6H6)(PCy3)(CO)RuH]+BF4- has been found to be an effective catalyst for the oxidative C-H coupling reaction of phenols with aldehydes to give 2-acylphenol compounds. The coupling of phenols with α,ß-unsaturated aldehydes selectively gives the flavene derivatives. The catalytic method mediates direct oxidative C-H coupling of phenol and aldehyde substrates without using any metal oxidants or forming wasteful byproducts.

A case of recurring acute exudative polymorphous vitelliform maculopathy successfully treated with intravitreal Ozurdex injection.

INTRODUCTION: We describe a case of acute exudative polymorphous vitelliform maculopathy (AEPVM) that recurred 9 years after the initial event. To the best of our knowledge, this is the first report of recurrent AEPVM showing recovery of retinal and retinal pigment epithelium (RPE) function and good visual outcome following treatment with intravitreal corticosteroid. CASE DESCRIPTION: A 45-year-old Caucasian woman first presented with AEVPM in 2009. Her condition spontaneously resolved and she remained stable over several years. 9 years later, her condition recurred with bilateral reduction in visual acuity. Fundus examination revealed multiple small yellowish subretinal lesions across the posterior pole in both eyes. Optical coherence tomography (OCT) showed bilateral cystoid macular oedema (CMO). She was referred for electrophysiology and her electrooculogram findings were in keeping with severe generalised RPE dysfunction bilaterally, with a light peak to dark trough ratio (Arden index) of 110%, comparable to her initial presentation 9 years earlier. She was initially treated with oral steroids with some improvement. However, the maculopathy in the left eye recurred on cessation of oral treatment. A sustained-release 700ug dexamethasone intravitreal implant (Ozurdex®) was inserted in the left eye to which she responded remarkably, with improvement in visual acuity and complete resolution of the CMO. A year later, at her most recent clinic visit in March 2021, there was no evidence of any further recurrence. CONCLUSION: Our case demonstrates clinical and imaging findings consistent with recurrence of AEPVM with CMO that has been successfully treated with Ozurdex®.

Prognostic Value of Optic Nerve Sheath Diameters after Acute Ischemic Stroke According to Slice Thickness on Computed Tomography.

The optic nerve sheath diameter (ONSD) can predict intracranial pressure and outcomes in neurological disease, but it remains unclear whether a small ONSD can be accurately measured on routine CT images with a slice thickness of approximately 4-5 mm. We measured the ONSD and ONSD/eyeball transverse diameter (ETD) ratio on routine-slice (4 mm) and thin-slice (0.6-0.75 mm) brain CT images from initial scans of acute ischemic stroke (AIS) patients. ONSD-related variables, National Institutes of Health Stroke Scale (NIHSS) scores, and age were compared between good (modified Rankin Scale [mRS] ≤ 2) and poor (mRS > 2) outcomes at discharge. Among 155 patients, 38 had poor outcomes. The thin-slice ONSD was different between outcome groups (p = 0.047), while the routine-slice ONSD showed no difference. The area under the curve (AUC) values for the ONSD and ONSD/ETD were 0.58 (95% CI, 0.49-0.66) and 0.58 (95% CI, 0.50-0.66) on the routine-slice CT, and 0.60 (95% CI, 0.52-0.68) and 0.62 (95% CI, 0.54-0.69) on the thin-slice CT. The thin-slice ONSD/ETD ratio correlated with initial NIHSS scores (r = 0.225, p = 0.005). After adjusting for NIHSS scores and age, ONSD-related variables were not associated with outcomes, and adding them to a model with NIHSS scores and age did not improve performance (all p-values > 0.05). Although ONSD measurements were not an independent outcome predictor, they correlated with stroke severity, and the thin-slice ONSD provided a slightly better prognostic performance than the routine-slice ONSD.

Safety Assessment and Evaluation of Probiotic Potential of Lactobacillus bulgaricus IDCC 3601 for Human Use.

Lactic acid bacteria (LAB) are probiotic microorganisms widely used for their health benefits in the food industry. However, recent concerns regarding their safety have highlighted the need for comprehensive safety assessments. In this study, we aimed to evaluate the safety of L. bulgaricus IDCC 3601, isolated from homemade plain yogurt, via genomic, phenotypic, and toxicity-based analyses. L. bulgaricus IDCC 3601 possessed a single circular chromosome of 1,865,001 bp, with a GC content of 49.72%, and 1910 predicted coding sequences. No virulence or antibiotic resistance genes were detected. Although L. bulgaricus IDCC 3601 exhibited antibiotic resistance to gentamicin and kanamycin, this resistance is an intrinsic feature of this species. L. bulgaricus IDCC 3601 did not produce biogenic amines and did not exhibit hemolytic activity. Phenotypic analysis of enzyme activity and carbohydrate fermentation profiles revealed the metabolic features of L. bulgaricus IDCC 3601. Moreover, no deaths or abnormalities were observed in single-dose oral toxicity tests, suggesting that L. bulgaricus IDCC 3601 has no adverse effect on human health. Finally, L. bulgaricus IDCC 3601 inhibited the growth of potential carbapenem-resistant Enterobacteriaceae. Therefore, our results suggest that L. bulgaricus IDCC 3601 is a safe probiotic strain for human consumption.

Simulation of a randomly percolated CNT network for an improved analog physical unclonable function.

Carbon nanotube networks (CNTs)-based devices are well suited for the physically unclonable function (PUF) due to the inherent randomness of the CNT network, but CNT networks can vary significantly during manufacturing due to various controllable process conditions, which have a significant impact on PUF performance. Therefore, optimization of process conditions is essential to have a PUF with excellent performance. However, because it is time-consuming and costly to fabricate directly under various conditions, we implement randomly formed CNT network using simulation and confirm the variable correlation of the CNT network optimized for PUF performance. At the same time, by implementing an analog PUF through simulation, we present a 2D patterned PUF that has excellent security and can compensate for error occurrence problems. To evaluate the performance of analog PUF, a new evaluation method different from the existing digital PUF is proposed, and the PUF performance is compared according to two process variables, CNT density and metallic CNT ratio, and the correlation with PUF performance is confirmed. This study can serve as a basis for research to produce optimized CNT PUF by applying simulation according to the needs of the process of forming a CNT network.

Gate Capacitance Coupling of Double-Gate Carbon Nanotube Network Transistors.

Carbon nanotube (CNT) network channels constructed using a high-purity CNT solution for use in CNT thin-film transistors have the advantages of the possibility of requiring a low-temperature process and needing no special equipment. However, there are empty spaces between individual CNTs, resulting in unexpected effects. In this study, double-gate (DG) CNT network transistors were fabricated and measured in four different configurations to observe the capacitive coupling effects between the top gate (TG) and bottom gate (BG) in the DG structure. As a result, the electrical characteristics measured with the BG with a thicker gate oxide while floating the TG were similar to those measured with the TG with a thinner gate oxide. A comparison of the measured transfer curves showed that TG and BG were strongly coupled through the empty spaces in the channels. In addition, we evaluated the capacitance coupling effect due to changes in the CNT density, which is closely related to the empty space of the network channel. Finally, we proposed a method to determine the effective gate capacitance by considering the empty spaces between CNTs, which enabled the accurate evaluation of mobility. The effects of these materials were demonstrated by fabricating transistors using Al2O3, HfO2, and ZrO2 as TG oxide materials. By focusing on considerations based on the properties of CNT materials, our study provides valuable insights into accurate electrical modeling and potential advancements in CNT-based devices.

Iron-Catalyzed, Directed Benzylic Borylation.

Herein, we describe an iron-catalyzed benzylic C-H borylation reaction. The reaction targets primary and secondary C(sp3)-H bonds to deliver high-value boronic esters under mild conditions with short (7-8 min) reaction times. Functional groups are well tolerated, and complete site selectivity is observed in the presence of multiple benzylic C-H bonds.

Outcomes of Descemet's membrane endothelial keratoplasty performed in combination with, before, or after cataract surgery in Fuchs' endothelial dystrophy: A review of the literature and meta-analysis.

This review aimed to compare the outcomes of Descemet's membrane endothelial keratoplasty (DMEK) in combination with (category 1), before (category 2), or after cataract surgery (category 3) in patients with Fuchs' endothelial dystrophy (FED). Primary outcome was gain in best-corrected log of minimum angle of resolution (logMAR) visual acuity (BCVA). Secondary outcomes were graft detachment, rebubbling rates, rejection, failure, and endothelial cell loss (ECL). In category 1, 2, and 3, 12 studies (N = 1932) were included (five in category 1 [n = 696], one in category 2 [n = 286], and two in category 3 [n = 950], and the remaining four compared between two of the three categories). At 6 months, the gain in BCVA was 0.34 ± 0.04, 0.25 ± 0.03, and 0.38 ± 0.03 logMAR in category 1, 2, and 3, respectively. The difference was significant between categories 1 and 2 (Chi2 = 11.47, P < 0.01) and categories 2 and 3 (Chi2 = 35.53, P < 0.01). At 12 months, the gain in BCVA was 0.52 ± 0.05 and 0.38 ± 0.06 logMAR in categories 1 & 3 (Chi2 = 14.04, P < 0.01). The rebubbling rates were 15%, 4%, and 10% (P < 0.01) and the graft detachment rates were 31%, 8%, and 13% (P < 0.01) in categories 1, 2, and 3, respectively. However, graft rejection, survival rates, and ECL at 12 months were not different between categories 1 and 3. There is low certainty evidence that gain in BCVA in category 1 was comparable to category 3 at 6 months; however, it was significantly better with category 3 at 12 months. Although rebubbling and graft detachment rates were highest in category 1, there was no significant difference in graft rejection, survival rates, and ECL. Further high-quality studies are likely to change the effect estimate and have an impact on the confidence of the estimate.