Clinical Chemistry and Laboratory Medicine (CCLM)

ObjectiveIn vitro cytokine release assays are essential tools in immunological research, yet their reliability can be influenced by various pre-analytical factors. This study was initiated to investigate the potential impact of lithium heparin whole blood collection tubes from different manufacturers on assay performance. The objective of this work is to urge the scientific community to consider whole blood collection tubes as a significant and often underestimated source of variability in cell-based assays. Results description210 whole blood samples from healthy donors were collected in plastic lithium heparin tubes sourced from 20 different suppliers worldwide. Testing revealed that 8 out of 20 of these tubes contained endotoxin levels >0.1{square}EU/mL, which triggered significant variability in cytokine responses (IFN-{gamma} and IL-6) in cell-based assays. A dose-response curve using LPS from E. coli confirmed the impact of endotoxins on interferon-{gamma} production.

A multivariant total subclass analysis has been performed for a control cohort (n=15) and a long COVID patient cohort (n=15) measuring the IgG1, IgG2, IgG3, IgG4 and IgE response to the following 14 variants of SARS-CoV-2: Wuhan, Alpha, Delta, BA.1, BA.2, BA.5, EG.5.1, XBB.1.5, BA.2.75, CH.1.1, BA.2.12.1, BQ.1.1, JN.1, and KP.3. Significant differences (p < 0.05 and p < 0.005) between concentrations of IgG subclasses by variant were found in 24% of variants and in mean-normalised distributions. The medians of the mean-normalised distributions were significantly lower for IgG1 (p < 0.05) in long COVID patients compared with controls, and significantly higher (p < 0.005) for levels of IgG3, IgG4 and IgE for long COVID patients. A preliminary diagnostics classification analysis performed by variation of the mean-normalised upper and lower percentiles symmetrically for IgG3 showed a long COVID diagnostic sensitivity of 80%, and specificity of 80% for the 60th percentile threshold of the control cohort. Three types of long COVID can be identified: patients with at least one variant below the threshold (hypo-immune), patients with at least one variant above the threshold (hyperimmune) and patients with IgG3 levels within the reference range. The multivariant subclass spectrum indicates IgG4 and IgE elevations due to potential chronic antigen exposure from persistent virus or autoimmunity and may indicate potential therapeutic interventions.

PurposeTo evaluate the diagnostic performance of anterior segment optical coherence tomography (ASOCT) compared to slit lamp examination for identification of corneal perforation in microbial keratitis, and to assess ASOCT grading reproducibility. MethodsWe conducted a diagnostic concordance study of 150 eyes with microbial keratitis at a tertiary eye hospital in India. Two masked graders independently evaluated ASOCT scans for perforation, with disagreements resolved by consensus. We calculated Cohens kappa for inter-grader concordance and intra-grader repeatability. Sensitivity and specificity of slit lamp examination were calculated using ASOCT as the reference standard. Logistic regression identified factors associated with disagreement between modalities. ResultsInter-grader agreement for ASOCT was near-perfect ({kappa}=0.98; 95% CI, 0.92-1.00). ASOCT identified perforation in 24 eyes (16.0%) compared to 12 eyes (8.0%) by slit lamp examination. Using ASOCT as reference, slit lamp examination demonstrated 33.3% sensitivity (95% CI, 14.9-52.2%) and 96.8% specificity (95% CI, 93.4-99.2%). Odds of disagreement were significantly higher for eyes with stromal thinning (OR=8.19; 95% CI, 2.27-29.54), mid-stromal involvement (OR=4.44; 95% CI, 1.08-18.30), and infection within 2mm of the limbus (OR=8.81; 95% CI, 1.77-43.80). ConclusionsASOCT enables highly reproducible perforation grading and detects substantially more perforations than slit lamp examination, particularly in severe disease. These findings support ASOCT as an objective tool for clinical assessment and outcome ascertainment in microbial keratitis.

ObjectiveDrug poisoning cases due to overdoses of over-the-counter (OTC) medications are increasing, and comprehensive measurement of blood drug concentrations, including OTC drugs, is important in emergency medicine and forensic science. In this study, we developed a simultaneous quantification method for blood drug levels using LC-MS/MS with solid-phase mini-cartridge SmartSPE for sample preparation. MethodsThe target analytes were acetaminophen, caffeine, flunitrazepam, 7-aminoflunitrazepam, risperidone, and phenobarbital. Internal standards (IS) used were acetaminophen-d4, caffeine-d9, diazepam-d5, and phenobarbital-d5. For solid-phase extraction, three types of Smart-SPE columns (AiSTI Science Co., Ltd.) were employed. 100 {micro}L of blank whole blood samples were subjected to protein precipitation using methanol and acetonitrile, and the supernatant obtained after centrifugation was processed using Smart-SPE for sample cleanup. Chromatographic separation was performed on a CAPCELL PAK INERT ADME-HR column (Osaka Soda), and detection was carried out in both positive and negative ESI modes. ResultsLinearity was observed for all drugs across the therapeutic to coma-death concentration ranges, with correlation coefficients exceeding 0.99 in all cases. Intra-day accuracy ranged from 98.06% to 110.87%, with precision between 0.19% and 17.37%. Inter-day accuracy ranged from 97.82% to 112.35%, with precision between 0.53% and 9.18%. Recovery rates varied from 72.9% to 95.1%, and matrix effects ranged from 91.2% to 113.0%. DiscussionThe validation results demonstrated satisfactory performance for all analytes, suggesting that this simultaneous quantification method may serve as a reliable analytical approach. Further investigations into sample stability are planned, and if analyses of actual specimens confirm its applicability, the method can be reported as a novel approach for simultaneous quantification of blood drug levels using a new sample pretreatment technique.

PurposeTo investigate whether the geometric shape of the globe, sagittal length (SL) and Posterior Radius of curvature (rPC), differ between patients with rhegmatogenous retinal detachment (RD) and macular hole (MH), and to determine if these parameters offer better diagnostic differentiation than axial length (AL) alone. MethodsThis retrospective study included 20 MH and 20 RD patients. Groups were axial-length matched. Ocular biometry was performed with ultrasound to measure AL and SL. The PR was calculated. Due to the non-normal distribution of the data, the Mann-Whitney U test was used for continuous variables and Fishers Exact Test for gender distribution. ResultsThere was no significant difference between the MH and RD groups in terms of age (median: 33.8 vs 32.6 years; p=1.00), gender (p=0.341), or AL (27.65 mm vs 28.58 mm; p=0.39). However, RD eyes showed significantly higher SL (median: 25.70 mm vs 22.90 mm; p=0.001) and a significantly flatter rPC (median: 28.25 mm vs 24.30 mm; p < 0.001). ConclusionRD eyes exhibit a distinct vertical equatorial expansion and posterior flattening that is not present in MH eyes, despite similar axial lengths. These geometric differences suggest that SL and PR are structural risk factors in vitreoretinal disease.

PurposeAqueous humor drains fluid from the eye not only via the conventional pathway through the trabecular meshwork and Schlemms canal, but also within the eye is known to occur via pathways through the posterior chamber and optic nerve to the cerebrospinal fluid (CSF) surrounding the optic nerve. The mechanism is poorly understood, and non-invasive method for evaluation in living humans has not been established. We previously showed that eye drops containing O-17-labeled water (H217O) distribute in the anterior chamber but not the vitreous. This study aimed to evaluate the distribution of H217O in the CSF along the optic nerve. MethodsFive ophthalmologically normal participants (20-31 years, all females) were selected from a previous prospective study based on 1H MR images of the eyes that included the optic nerve. They received eye drops of 10 mol% H217O in their right eye. Dynamic image time series was created by normalizing the signal of each 1H-T2WI by the pre-drop average signal. Region-of-interest analyses were performed for signal changes in the anterior chamber, vitreous, and CSF. ResultsIn the quantitative evaluation, the normalized intensity in the anterior chamber and CSF was significantly lower than that in the pre-drop signal (anterior chamber: 0.78 {+/-} 0.07, p < 0.005; CSF: 0.89 {+/-} 0.07, p < 0.05). No distribution was identified in the vitreous. Qualitatively, the distribution of H217O in the anterior chamber was detected in all five participants and in the CSF of four participants (80%). ConclusionH217O eye drops were distributed in the anterior chamber and CSF, but not in the vitreous. These findings suggest that the visualization of aqueous humor outflow, not via the Schlemms canal, may contribute to ocular fluid homeostasis, including the ocular glymphatic system.

BackgroundThere is a need for synthetic peptide-based serologic assays that exploit avidity to replace whole antigens while enabling low-cost diagnostics in resource-limited settings. ObjectiveTo evaluate the diagnostic accuracy of a polymeric peptide-based ELISA leveraging avidity to enhance signal. MethodA 15-member SARS-CoV-2 peptide library corresponding to multiple epitope clusters and proteins was screened by indirect ELISA using pooled sera from RT-PCR-confirmed COVID-19 patients to identify peptides with possible diagnostic utility. The identified lead candidate, S559, possessed terminal cysteine-substitution to allow disulfide polymerization, and the resulting avidity gain was evaluated by comparing the apparent dissociation constant (KDapp) before and after depolymerization with N-acetylcysteine. The performance of an optimized ELISA using S559 was evaluated on 1,222 prospectively collected COVID-19 serum samples and 218 biobanked pre-COVID control serum samples. ResultsPolymeric S559 with a KDapp of 29.26 nM-1was demonstrated to have a 218% avidity gain relative to the completely depolymerized form. At pre-defined thresholds, the optimized S559 ELISA has a sensitivity and specificity of 83.39% (95%CI: 81.18% and 85.43%) and 96.79% (95%CI: 93.50% and 98.70%), respectively. At post hoc thresholds determined by Youden index, sensitivity and specificity reached 95.01 (95% CI: 93.63% - 96.16%) and 100.00% (95% CI: 98.32% - 100.00%), respectively. ConclusionHomomultivalent epitope presentation using polymeric S559 allows a highly specific immunoassay using human sera that may have important value in detecting antibodies, whether for diagnosing infection, confirming vaccination status or conducting surveillance.

PurposeMuller cell (MC) morphology and markers were investigated using histology and immunohistochemistry in an eye with clinically documented multifocal geographic atrophy (GA) and correlated with clinical images. MethodsThe donor was followed clinically for five years and last examined six years before death. The superior posterior pole retina was dissected and immunolabeled with antibodies against glial fibrillary acidic protein (GFAP; activated MCs and astrocytes) and glutamine synthetase (GS, MC) and Ulex Europaeus Agglutinin-1 lectin (blood vessels) before embedding for JB-4 cross section analysis. The inferior macula was cryopreserved. Cryosections were immunolabeled with MC homeostatic and activation markers. Transmission electron microscopy (TEM) of the fellow eye was used to study ultrastructure changes. ResultsGross examination demonstrated mottled retinal pigment epithelium (RPE) over presumably calcified drusen. In the submacular retina, MC processes surrounding both drusen and outer retinal pigmented lesions created a large subretinal membrane. Cryosection analysis demonstrated persistence of aquaporin 4 and GS in MCs with both proteins prominently expressed in the subretinal membrane. Increased MC S100B and GFAP expression were also observed in the atrophic area as well as the OJZ. Cryosection labeling and TEM confirmed the MC encasing calcified drusen and RPE debris as well as invading basal laminar deposits. ConclusionsThis multifocal GA case demonstrates how MC activation and structural changes surrounding individual drusen could coalesce, contributing to photoreceptor loss. MCs penetrating basal laminar deposits and encasing calcified drusen suggests that they are attempting to clear these and/or protect the retina from harmful contents.

PurposeTo evaluate the intra- and inter-grader concordance of anterior segment optical coherence tomography (ASOCT) grading for detection of endothelial plaque in microbial keratitis, and to compare endothelial plaque detection via ASOCT grading versus in-person slit lamp examination. MethodsDiagnostic concordance study of 150 consecutive patients with microbiologically confirmed bacterial or fungal keratitis at a high-volume tertiary eye hospital in India. Two masked ophthalmologist graders independently evaluated ASOCT images for presence of two morphologically distinct endothelial plaque subtypes noted during image review (round and flat plaques). We assessed intra-grader and inter-grader concordance for each endothelial plaque morphology and for presence of either morphology. Diagnostic agreement between ASOCT and in-person slit lamp examination was evaluated using percent agreement, Cohens kappa, sensitivity, and specificity. Univariable and multivariable logistic regression was used to assess odds of disagreement between ASOCT and slit lamp examination for endothelial plaque detection. ResultsASOCT detection showed near perfect inter-grader agreement for round endothelial plaques (kappa 0.88, 94.7% agreement), flat endothelial plaques (kappa 0.84, 92.0% agreement), and either plaque (kappa 0.88, 94.0% agreement). Intra-grader agreement was substantial to near perfect for both graders across all plaque types (kappa 0.70-0.86). Ophthalmologist slit lamp examination identified endothelial plaque in 6.0% eyes, while ASOCT detected round plaques in 32.7%, flat plaques in 43.3%, and either plaque in 55.3% of eyes. Using ASOCT as reference, slit lamp examination demonstrated sensitivity of 16.3% for round plaques, 6.2% for flat plaques, and 9.6% for either plaque, with specificity exceeding 94% for all. Poor visual acuity (logMAR [&ge;]1.0) was associated with increased disagreement for round plaques (adjusted OR 5.04), flat plaques (adjusted OR 3.63), and either plaque (adjusted OR 3.98). Bacterial infection was associated with increased disagreement for any endothelial plaque (adjusted OR 4.56). ConclusionSlit lamp examination substantially under-detects endothelial plaque compared to ASOCT, while ASOCT enables reproducible detection with excellent intra- and inter-grader agreement. These findings support incorporation of ASOCT imaging into microbial keratitis evaluation protocols. Differences in round and flat endothelial plaque morphologies warrant further investigation.

PurposeTo evaluate the efficacy and safety of Gene III L-ergothioneine (EGT) Eye Care Solution for alleviating ocular discomfort in patients with dry eye syndrome (DES). MethodsIn this single-center, randomized, open-label, self-controlled trial, 40 DES patients were randomly assigned in a 1:1 ratio to one of two treatment groups: Group 1 received the Gene III EGT Eye Care Wash Solution via an eye wash cup (5 mL/vial), and Group 2 received the same formulation as Gene III EGT Eye Care Drops Solution (0.5 mL/vial). Outcomes were assessed at baseline and after 14 days using validated scales, including the Chinese Dry Eye Questionnaire, the Ocular Surface Disease Index (OSDI), and the Comprehensive Assessment of Visual Fatigue (CAVF), along with fluorescein tear film break-up time (TBUT). ResultsCombined analysis (N = 40) demonstrated significant improvements in dry eye symptoms with mean scores decreasing from 12.50 to 10.65 (P = 0.0353). Ocular Surface Disease Index (OSDI) scores improved from 12.25 to 9.2 (P = 0.0309), and visual fatigue, assessed by the CAVF scale, decreased from 11.18 to 6.60 (P = 0.0008). Significant increases in TBUT were observed in both groups, with left eye first TBUT rising in Group 1 (P = 0.0199) and Group 2 (P < 0.0001). No treatment-related adverse events were reported. ConclusionGene III EGT Eye Care Solution effectively alleviated DES symptoms and demonstrated a favorable safety profile. Larger placebo-controlled trials are warranted to confirm these findings.

PurposeTo evaluate the three-year efficacy and safety of compound trabeculectomy for uveitic glaucoma (UG). MethodsThis retrospective study enrolled 51 patients (53 eyes) requiring compound trabeculectomy, divided into UG (25 eyes) and non-UG groups (28 eyes). Outcomes including intraocular pressure (IOP), medication use, surgical success rates, and complications were analyzed over 3 years. ResultsBaseline characteristics including age, sex, preoperative IOP and medication use were comparable (all P>0.05). At 36 months, postoperative IOP was showed no significant differences, which was 15.4{+/-}8.4 mmHg and 14.6{+/-}3.3 mmHg (P=0.73) with 54% and 55% reduction (P=0.88) in UG and non-UG groups respectively. The qualified success rate was 76.0% and 85.7% at 36 months in UG and non-UG group, and Kaplan-Meier analysis showed no significant difference. Medication reduction of UG group was significant lower than non-UG group (P=0.0058). Comparable complication rates were observed between groups (all P>0.05), yet bleb scarring and cataract progression showed elevated incidence in both cohort. ConclusionCompound trabeculectomy effectively reduced IOP and medications use in UG and non-UG. There was no significant difference in both qualified and completed success rate between UG and non-UG. Complications of filtering bleb fibrosis and cataract progression should be pay close attention for both groups.

BackgroundSevere Fever with Thrombocytopenia Syndrome (SFTS) is an acute infectious disease with high mortality. This study aimed to develop a quantitative scoring system for grading SFTS severity using dynamic clinical data. MethodsA retrospective study included 547 confirmed SFTS patients from two hospitals. Clinical data were collected over a 14-day course (divided into four phases). Patients were grouped into survivors (n=451) and non-survivors (n=96). Statistical analyses, including Kaplan-Meier curves and log-rank tests, were performed. An external validation cohort of 44 new patients was used to validate the scoring system via C-statistic, calibration curves, and decision curve analysis (DCA). ResultsOf 547 patients, 96 (17.55%) were non-survivors. Multivariate logistic regression identified six independent prognostic factors across phases: age, platelet (PLT), aspartate aminotransferase (AST), and creatinine (Cr) (days 5-7); age, red blood cell distribution width (RDW), Cr, and lactate dehydrogenase (LDH) (days 8-10); Cr and LDH (days 11-14). A scoring system (0-11 points) was developed, stratifying patients into low (0-3), intermediate (4-7), and high (8-11) risk groups, with adverse outcome rates of 1.04%, 22.92%, and 76.04%, respectively. Kaplan-Meier curves showed significant prognostic differences (log-rank P<0.001). External validation (44 cases) confirmed excellent performance: AUC 0.810-0.952, good calibration (Hosmer-Lemeshow P>0.05), and net clinical benefit (DCA Eavg 0.068-0.098, Emax 0.422-0.559). ConclusionA dynamic SFTS severity scoring system was developed and validated. Internal and external validation confirmed its reliability and clinical utility, providing a simple, practical tool for timely assessment and early intervention.

Influenza A, Influenza B, SARS-CoV-2, Respiratory Syncytial Virus and other respiratory pathogens are an ongoing public health concern. The ability to rapidly identify these viruses early in infection is essential for effective treatment and outbreak control. The AMDI Fast PCR Mini Respiratory Panel (MRP) incorporates sample preparation and real-time RT-PCR for detection of Flu A, Flu B, SARS-CoV-2 and RSV from anterior nasal swab (ANS) specimens in less than 10 minutes at the point of care. We established the analytical performance characteristics of the Fast PCR MRP and determined that the limit of detection (LoD) is 250 copies/mL for Flu A and RSV, and 500 copies/mL for Flu B and SARS-CoV-2. In a reproducibility study at 3 clinical sites, there was at least 98.2% positivity for each target for a weak positive (2x LoD) sample, at least 99.6% positivity for a moderate positive (5x LoD) sample and the negative sample returned a negative result for at least 99.3% of the tests. Fast PCR MRP had 100% analytical reactivity to all strains tested (23 Flu A, 6 Flu B, 8 SARS-CoV-2 and 6 RSV) and at least 98% predicted inclusivity from in silico analysis. There was no cross-reactivity to 40 viruses, bacteria and fungi, nor interference for 15 endogenous and exogenous substances in ANS matrix. The Fast PCR MRP delivers excellent analytical performance comparable to high complexity laboratory assays, at the point of care.

AimsPrimary objective: To determine the minimum inhibitory concentration (MIC) of key antifungal drugs (natamycin, amphotericin B, voriconazole and econazole) against fungal isolates cultured from fungal keratitis patients in South India. Secondary objective: to ascertain correlations between antifungal resistance and patient outcome. MethodsIn this prospective observational study, MIC values were determined for fungal isolates cultured from 153 patients, with samples collected between May - August 2025. Clinical characteristics were collected at baseline, one-week and one-month following enrolment to the study. Mean antifungal MIC per fungi genera were compared. Statistical differences in MIC and patient characteristics were determined via multiple logistic or linear regression. Significance for participant outcome against resistant/non-resistant fungi were determined by Fishers exact test. ResultsResistance of Fusarium spp. isolates to: natamycin: 38.3%; amphotericin B: 93.8%; voriconazole: 97.5% and econazole: 76.5%. Resistance of Aspergillus spp. isolates to: natamycin: 66.7%; amphotericin B: 87.9%; voriconazole: 6.1%; and none were resistant to econazole. Natamycin MIC correlated with worse baseline (P[&le;]0.01) and one-week (P[&le;]0.05) visual acuity and ulcer severity. Poor patient outcomes (non-healing or therapeutic keratoplasty) were elevated 6.5x where the infection was caused by natamycin resistant Aspergillus, compared to sensitive Aspergillus strains (P[&le;]0.05). ConclusionThe majority of fungal isolates were resistant to multiple antifungals, none of the Fusarium isolates were sensitive to all four drugs, and 15% were resistant to all four drugs. Aspergillus isolates had high levels of resistance to the polyenes, but remained largely susceptible to the azoles. Overall, worse patient outcomes were associated with increased natamycin MIC. Key MessagesO_ST_ABSWhat is already known on this topicC_ST_ABSFungal keratitis is a major cause of blindness worldwide, disproportionately affecting those across the tropics, with incidence increasing across temperate climates. The majority of cases are caused by the filamentous fungi Fusarium spp. and Aspergillus spp.. Antifungal resistance is poorly characterised in fungal keratitis. What this study addsWe report the fungal aetiology and the minimum inhibitory concentration (MIC) against natamycin, amphotericin B, voriconazole and econazole of isolates cultured from 153 patient corneal scrape samples between May - August 2025 at a South Indian hospital. We found high levels of fungal resistance, with Fusarium isolates having high levels of resistance to both polyenes and azoles. Aspergillus isolates showed good azole sensitivity, but high levels of resistance to polyenes. Aspergillus resistance to natamycin correlated with worse clinical outcomes at one-month. Natamycin resistance contributed to worse visual acuity and ulcer severity at baseline and one-week follow-up across all fungi. How this study might affect research, practice or policyOur study confirmed that natamycin was best available first-line treatment for Fusarium. Aspergillus isolates were mostly resistant to natamycin and amphotericin B, and this impacted patient outcomes. SynopsisWe identified high incidence of multi-drug resistant fungi, and that patients were more likely to have a poor clinical outcome if the fungal isolate was resistant to natamycin. This was most pronounced for Aspergillus isolates.

BackgroundDiabetes mellitus (DM) remains a major global health challenge and is associated with vision-threatening complications, including diabetic macular edema (DME), a leading cause of visual impairment. Dyslipidemia has been implicated in the development of macular edema through mechanisms involving vascular permeability, endothelial dysfunction, and chronic inflammation. However, evidence regarding the relationship between lipid abnormalities and macular edema remains inconsistent across studies. AimThis study aimed to evaluate the association between abnormal lipid profiles and diabetic macular edema among patients with type 2 diabetes mellitus attending Kilimanjaro Christian Medical Centre (KCMC). MethodsA hospital-based analytical cross-sectional study was conducted among 296 diabetic outpatients at KCMC. Participants underwent comprehensive ophthalmic evaluation including fundoscopy and imaging with optical coherence tomography (OCT) for assessment of macular edema. Blood samples were collected for biochemical lipid analysis. Data were cleaned and analyzed using STATA version 17. ResultsDiabetic macular edema was identified in 56.4% (167/296) of participants. Abnormal lipid parameters were common, with elevated total cholesterol observed in 48.6%, triglycerides in 43.6%, low-density lipoprotein (LDL) in 36.1%, and reduced high-density lipoprotein (HDL) in 38.9% of patients. Elevated total cholesterol, triglycerides, and LDL levels showed significant associations with macular edema (p < 0.05). After multivariable adjustment, serum triglycerides remained independently associated with macular edema (p = 0.002). ConclusionDyslipidemia demonstrated a significant association with diabetic macular edema, with serum triglycerides emerging as an independent predictor. These findings highlight the importance of lipid monitoring, lifestyle modification, and strengthened screening strategies in reducing the burden of vision-threatening diabetic complications.

PurposeTo investigate the epidemiology of post-injection endophthalmitis (PIE) and evaluate the association of sociodemographic and clinical factors with incidence, timing of onset, and presenting visual acuity (VA) using the IRIS(R) Registry (Intelligent Research in Sight). DesignRetrospective cohort study. ParticipantsPatients with endophthalmitis after an intravitreal anti-VEGF injection in the IRIS(R) Registry from 2016-2023. MethodsOnly the first anti-VEGF injection per eye was included. Exclusion criteria were cataract surgery during the study, intravitreal corticosteroids within 30 days prior to PIE, uveitis, or cystoid macular edema. Mean best VA was recorded within 100 days prior to anti-VEGF treatment and at the time of PIE. Regression modeling evaluated associations between endophthalmitis and sociodemographic and clinical factors, and time to PIE. Linear regression assessed predictors of VA at the time of PIE, and descriptive statistics were used to analyze time to onset. Main Outcome MeasuresIncidence of post-injection endophthalmitis, time to symptom onset, and best VA at diagnosis. ResultsAmong 1,025,788 eyes treated, 600 (0.059%) developed endophthalmitis. Key risk factors included residence in U.S. territories (OR = 2.62; P = 0.038 vs. Northeast) and history of intravitreal corticosteroid injection (OR = 2.35; P = 0.004 vs. no history). The strongest protective factor was non-smoking (OR = 0.71; P = 0.023 vs. smokers). The median time from injection to onset of PIE was 5 days (interquartile range [IQR]: 3-8). The salient predictors of time to PIE included patient age (4.3 days sooner per decade older; P = 0.04), prior corticosteroid treatment (11.7 days sooner; P = 0.02), and a diagnosis of diabetic retinopathy (2.3 days sooner; P=0.03). Baseline VA before PIE was the only significant predictor of VA at the time of PIE diagnosis (0.67, P < 0.001). ConclusionPost-injection endophthalmitis was significantly associated with residence in U.S. territories and prior intravitreal corticosteroid exposure, while non-smoking status was protective. Most cases presented 3-8 days following anti-VEGF injection. Older age, history of prior corticosteroid treatment, and diabetic retinopathy were associated with earlier PIE. Baseline VA predicted VA at the time of PIE.

Inter-alpha Inhibitor Proteins (IAIP) are serine protease inhibitors and a reliable inflammatory biomarker. We have demonstrated that IAIP levels decrease during sepsis in humans and animal models. Currently, enzyme-linked immunosorbent assay (ELISA) is the standard procedure to measure IAIP levels. We developed a lateral flow immunoassay (LFIA) that allows rapid, point of care detection. We compared IAIP levels in infants with sepsis and/or necrotizing enterocolitis (NEC) by ELISA and LFIA in a multi-center, cross-sectional study. Blood samples were collected from 47 infants with sepsis, 31 sepsis case controls, 52 gestational age (GA)-matched controls and 10 infants with culture-negative sepsis ("clinical sepsis"). We also collected samples from 17 infants with NEC, 7 NEC case controls and 15 GA controls. IAIP levels at presentation of acute events and over the next 72 hours were significantly reduced in infants with sepsis, NEC and culture negative sepsis when compared to controls. IAIP levels did not differ in patients with sepsis or culture negative sepsis. IAIP levels measured by ELISA and LFIA were highly correlated (R2 = 0.9326) and both showed reliable detection of neonatal sepsis, NEC and culture negative sepsis. IAIP levels were 80.0% sensitive and 92.3% specific using LFIA for the detection of neonatal sepsis. For detection of NEC, IAIP levels were 84.6% sensitive and 86.7% specific.

PurposeQuantitative metrics obtained from retinal fundus images (such as vessel length, tortuosity and other scale-dependent measures) are increasingly used as potential biomarkers for systemic diseases, including cardio- and neurovascular conditions. However, with the increasing prevalence of myopia and related axial growth, this study aims to evaluate if axial length scaling significantly alters the overall distributions of the inferred biomarkers when compared to biomarker data obtained without axial length scaling and if these effects can be corrected. Methods2,309 clinic visits from patients aged [&le;]21 years were analysed and extracted for axial-length scaling analysis (range) 20 to 28 mm). The retinal fundus photographs were automatically segmented using Automorph to extract biometric data, including vascular metrics. The parameters were further corrected for axial length using correction factors based on the Bennett-Littmann formula and true axial length. ResultsAxial length significantly influenced biometric parameters (vessel metrics) derived from fundus photography. The magnitude of error in diameter and length of blood vessels was approximately 4-5% for each 1 mm deviation from the reference axial length of 24 mm, whereas the error in vessel area was approximately 9-10% per 1 mm, consistent with the geometric expectation that area scales with the square of linear dimensions. The scaling corrections for different axial lengths are presented. ConclusionsAxial-length-related magnification introduces systematic bias into retinal vascular metrics from fundus photographs. Bennett-Littmann correction using true axial length reduces these errors and should be adopted in quantitative fundus imaging and Al biomarker development.

IntroductionAnti-vascular endothelial growth factor (anti-VEGF) therapies are standards of care for vision-threatening retinal diseases. This retrospective observational study describes demographics, utilization, best recorded visual acuity (BRVA), and safety among eyes with neovascular age-related macular degeneration (nAMD), diabetic retinopathy (DR), diabetic macular edema (DME), or retinal vein occlusion (RVO) treated with the biosimilar aflibercept-ayyh (PAVBLU(R)) in routine clinical practice. MethodsElectronic medical records from the Retina Consultants of America database of patients receiving aflibercept-ayyh (12/1/2024-10/31/2025) were analyzed, focusing on eyes with [&ge;]84 days of follow-up. The index date was the first documented aflibercept-ayyh injection. Postindex data were used to assess treatment patterns, BRVA (Wilcoxon signed rank test), and adverse events of special interest (AESIs). ResultsA total of 1,000 consecutive eyes from 989 patients received 3,730 injections of aflibercept-ayyh; most (91%) switched from prior anti-VEGF therapy and 9% were anti-VEGF treatment-naive. Disease distribution was 58% nAMD, 19% RVO, 16% DME, and 7% DR. Among switchers, median (IQR) number of prior injections was 21 (8-46). Median (IQR) follow-up was 6.0 months (4.6-7.1). Median (IQR) number of aflibercept-ayyh injections per eye was 4 (3-5). Among eyes with [&ge;]84 days of follow-up (n=889), mean BRVA expressed as logarithm of minimum angle of resolution (logMAR) remained stable for switchers (0.4 to 0.4; P=0.96) and improved from baseline in anti-VEGF-naive eyes (0.5 to 0.4; P<0.01). Confirmed AESIs included iritis (n=2; 0.05% of injections), with no events of vitreous cells, endophthalmitis, retinal detachment, retinal vasculitis, or vitreous hemorrhage. ConclusionIn this descriptive real-world analysis, aflibercept-ayyh was associated with stable visual acuity in previously treated eyes and vision improvement in treatment-naive eyes, with no new or unexpected safety findings, consistent with expectations for aflibercept. These findings add real-world experience to preexisting evidence demonstrating no clinically meaningful differences between aflibercept-ayyh (PAVBLU(R)) and reference aflibercept (EYLEA(R)). KEY SUMMARY POINTSO_ST_ABSWhy carry out this study?C_ST_ABSO_LIThe anti-vascular endothelial growth factor (VEGF) drug aflibercept, approved in 2011 and marketed in the United States as EYLEA(R),* has demonstrated efficacy in treating retinal diseases such as neovascular age-related macular degeneration (nAMD), diabetic retinopathy (DR), diabetic macular edema (DME), or retinal vein occlusion (RVO) and is a standard of care for these disorders. C_LIO_LIAflibercept-ayyh is a biosimilar to aflibercept that has demonstrated comparable efficacy and safety in the treatment of nAMD in a randomized controlled clinical trial. C_LIO_LIThis study describes the real-world use patterns, vision outcomes, and safety of aflibercept-ayyh in clinical settings in the United States for the treatment of nAMD, DR, DME, and RVO. C_LI What was learned from the study?O_LIIn this real-world study of 1,000 consecutive eyes treated with the biosimilar aflibercept-ayyh in patients with retinal diseases, we observed no new safety concerns and that aflibercept-ayyh maintained visual acuity in eyes switching anti-VEGF agents and improved vision in anti-VEGF-naive eyes, consistent with expected responses to aflibercept. C_LIO_LIThese findings support aflibercept-ayyh as a suitable treatment option when anti-VEGF therapy is indicated. *EYLEA(R) is a registered trademark of Regeneron Pharmaceuticals, Inc. PAVBLU(R) is a registered trademark of Amgen Inc. C_LI

The rapid diagnosis of Campylobacter infections is important for the management of infectious gastroenteritis. Although stool culture is considered the gold standard, its sensitivity is limited and it requires prolonged incubation times. We performed a prospective multicenter study at nine healthcare facilities in Japan to evaluate a Campylobacter rapid antigen test using stool specimens between March 2024 and August 2025. Patients with suspected infectious gastroenteritis were consecutively enrolled and tested using QuickNavi-Campylobacter and compared with the FilmArray Gastrointestinal Panel as the reference method. Discordant results were further evaluated by culturing and additional PCR assays. In total, 410 patients were included in the final analysis. The positive, negative, and total concordance rates between QuickNavi-Campylobacter and FilmArray Gastrointestinal Panel were 79%, 99%, and 93%, respectively. The positive concordance rate decreased in specimens collected [&ge;] 6 days after the onset of symptoms (50%). QuickNavi-Campylobacter demonstrated relatively good concordance with the FilmArray Gastrointestinal Panel in a real-world multicenter setting. These results suggest that this rapid antigen test may be particularly useful for the early diagnosis of suspected campylobacteriosis.