Changes In Cell Density Research Articles

Effects of Cumulative Dissipated Energy on Corneal Endothelial Cells in Phacoemulsification Based on Nucleus Sclerosis

Purpose: To investigate changes in corneal endothelial cell (CEC) by cumulative dissipated energy (CDE) during phacoemulsification.Methods: Based on the degree of nucleus sclerosis (NS), changes in CECs were compared preoperatively and at 1 and 3 months postoperatively in 67 eyes that underwent phacoemulsification.Results: The mean CDE used during surgery was 4.30 ± 2.31. A comparison of the surgical measurements before and 1 month after surgery revealed significant differences in the cell density (CD) and coefficient of variation (CV) (<i>p</i> < 0.001, 0.011, respectively). The CD showed significant differences among NS grades 2–6 (<i>p</i> < 0.001). The CDE increased significantly with higher NS grades (r = 0.809, <i>p</i> < 0.001). Within the same NS grade, there was a positive correlation between higher CDE and greater CEC loss (r = 0.559, <i>p</i> = 0.001). CD changes were significantly associated with increasing CDE (r = 0.612, <i>p</i> < 0.001). The CD loss also increased significantly from NS2 to NS6 (<i>p</i> < 0.001). At 3 months postoperatively, surgical measurements revealed a significant decrease in CD with increasing NS grade (<i>p</i> = 0.010).Conclusions: CDE increased with higher NS grades and there was a positive correlation between CDE and CEC loss. Therefore, surgeons should plan surgical techniques preoperatively to minimize CDE as NS increases. Additionally, it is important to assess CEC damage postoperatively based on surgical measurements in patients with high intraoperative CDE.

Findings in Corneal Endothelium by Confocal and Specular Microscopy in Patients with Fuchs Uveitis Syndrome Undergoing Phacoemulsification

ABSTRACT Purpose To compare corneal endothelial changes in patients with Fuchs Uveitis Syndrome (FUS) undergoing phacoemulsification surgery using confocal and specular microscopy. Methods We included 14 patients with unilateral FUS and cataracts who underwent phacoemulsification surgery in a Mexican referral center for inflammatory eye diseases. Preoperative confocal and specular microscopies were conducted, establishing baseline images for subsequent analyses. Surgery on the FUS eye was performed by a single surgeon and an intraocular lens was implanted in all cases. Both specular and confocal microscopy were repeated 6 months after FUS eye surgery and compared with baseline images. We used Image J to do a manual segmentation of KP and determine their density for further analysis hence developing a new tool for confocal microscopy image analysis. Results All patients underwent uneventful phacoemulsification surgery in the FUS eye. There was no significant change in endothelial cell density (ECD) from 2257 (±508.2) cells/mm2 preoperatively to 2214 (±535.1) cells/mm2 postoperatively (p = 0.809). Confocal microscopy revealed a decrease in Keratic Precipitate Density (KPD) from a median of 1413 (±2809.7) KPs/mm2 preoperatively to a median of 685.5 (1527.9) KPs/mm2 postoperatively (p = 0.036). Conclusions Phacoemulsification surgery in patients with FUS produces no significant loss of endothelial cells and morphological changes that can be detected by confocal and specular microscopy. We found a reduction in KPD 6 months after surgery on confocal microscopy. Additionally, our manual segmentation technique for KPs utilizing Image J offers a novel and practical approach for confocal microscopy image analysis.

Changes and Influencing Factors of Corneal Endothelial Cells and Central Corneal Thickness after Pediatric Cataract Surgery.

This study aimed to examine the impact of cataract surgery on the corneal endothelium and central corneal thickness (CCT) in pediatric patients, and to identify the factors associated with corneal alterations. This retrospective study included consecutive children undergoing bilateral or unilateral cataract surgery and intraocular lens (IOL) implantation at the Eye Hospital of Wenzhou Medical University, with or without posterior capsulorhexis and anterior vitrectomy, and an age-matched normal control group. This study aimed to assess whether changes in corneal parameters, including CCT, corneal endothelial cell density (CD), average cell area (AVE), standard deviation of size (SD), coefficient of variation (CV), percentage of hexagonal cells (6A) before and after surgery, and endothelial cell loss (ECL) differed among the bilateral cataract, unilateral cataract, and control groups. Furthermore, the potential effects of anterior vitrectomy, axial length (AL), preoperative anterior chamber depth (ACD), surgical duration, horizontal corneal diameter (HCD), intraoperative pupil diameter (PD), and the number of corneal sutures on corneal endothelial parameters and CCT were investigated. A total of 107 eyes from 107 children were included in the study. In the bilateral cataract group, CD significantly decreased, AVE and CCT significantly increased, and ECL was significantly higher than in the control group. The unilateral cataract group also exhibited a significant increase in CCT. Additionally, the numberofcornealsutures was negatively correlated with CD, and PD was negatively correlated with CV in the unilateral cataract group. Cataract surgery in pediatric patients results in increased CCT, reduced CD, and morphological changes in corneal cells. A greater number of corneal sutures and a smaller PD increased the risk of CD reduction and elevated CV in the unilateral cataract group, underscoring the need for ophthalmologists to minimize corneal damage in these children.

Free-swimming bacteria transcriptionally respond to shear flow

Surface-attached cells can sense and respond to shear flow, but planktonic (free-swimming) cells are typically assumed to be oblivious to any flow that carries them. Here, we find that planktonic bacteria can transcriptionally respond to flow, inducing expression changes that are beneficial in flow. Specifically, we use microfluidic experiments and quantitative modeling to show that in the presence of flow, planktonic Pseudomonas aeruginosa induce shear rate-dependent genes that promote growth in low-oxygen environments. Untangling this mechanism revealed that in flow, motile P. aeruginosa spatially redistribute, leading to cell density changes that activate quorum sensing, which in turn enhances the oxygen uptake rate. In diffusion-limited environments, including those commonly encountered by bacteria, flow-induced cell density gradients also independently generate oxygen gradients that alter gene expression. Mutants deficient in this flow-responsive mechanism exhibit decreased fitness in flow, suggesting that this dynamic coupling of biological and mechanical processes can be physiologically significant.

Intrapulmonary T Cells Are Sufficient for Schistosoma-Induced Pulmonary Hypertension.

Schistosomiasis is a parasitic infection that can cause pulmonary hypertension (PH). Th2 CD4 T cells are necessary for experimental Schistosoma-PH. However, if T cells migrate to the lung to initiate, the localized inflammation that drives vascular remodeling and PH is unknown. Mice were sensitized to Schistosoma mansoni eggs intraperitoneally and then challenged using tail vein injection. FTY720 was administered, which blocks lymphocyte egress from lymph nodes. T cells were quantified using flow cytometry, PH severity via heart catheterization, and cytokine concentration through ELISA. FTY720 decreased T cells in the peripheral blood, and increased T cells in the mediastinal lymph nodes. However, FTY720 treatment resulted in no change in PH or type 2 inflammation severity in mice sensitized and challenged with S. mansoni eggs, and the number of memory and effector CD4 T cells in the lung parenchyma was also unchanged. Notably, intraperitoneal Schistosoma egg sensitization alone resulted in a significant increase in intravascular lymphocytes and T cells, including memory T cells, although there was no significant change in parenchymal cell density, IL-4 or IL-13 expression, or PH. Blocking T cell migration did not suppress PH following Schistosoma egg challenge. Memory CD4 T cells, located in the lung intravascular space following egg sensitization, appear sufficient to cause type 2 inflammation and PH.

Pain perception, aqueous humour cytokines, and miosis response following first and second eye femtosecond laser-assisted cataract surgery: A randomised, prospective, intraindividual study.

To compare pain perception, pupil behaviours, and cytokine levels during first-eye and second-eye femtosecond laser-assisted cataract surgery (FLACS) and determine which is better regarding a short or long interval for bilateral FLACS. Notably, 96 eyes of 48 patients with bilateral cataracts underwent the first surgeries in the left or right eye, according to a random sequence. They were further randomised into 2- and 6-week subgroups based on surgery intervals. Pupil size was measured from captured images, and pain perception was assessed using a visual analog scale (VAS). Aqueous humour prostaglandin E2 (PGE2), monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6, and IL-8 levels were also quantitatively analysed. All patients were followed for 1 week to evaluate changes in endothelial cell density (ECD), central corneal thickness (CCT), and macular central subfield thickness (CST). Ocular pain was significantly higher in patients who underwent second-eye FLACS. First tear break-up time was negatively correlated with VAS score. MCP-1 levels were significantly higher in patients who underwent second-eye FLACS, and VAS scores were positively correlated with MCP-1 levels across all patients. There were no differences between sequential FLACS in miosis, PGE2, IL-6, IL-8 levels and changes in postoperative ECD, CCT, and CST. Patients who underwent second-eye FLACS after 6 weeks showed more CCT, CST, and MCP-1 changes than baseline. Second-eye FLACS causes more pain and upregulated MCP-1. There was no difference between sequential FLACS in PGE2 levels, miosis, and postoperative inflammation. Furthermore, first-eye FLACS triggered a sympathetic irritation, particularly after a 6-week interval.

Phenotypic memory in quorum sensing.

Quorum sensing (QS) is a regulatory mechanism used by bacteria to coordinate group behavior in response to high cell densities. During QS, cells monitor the concentration of external signals, known as autoinducers, as a proxy for cell density. QS often involves positive feedback loops, leading to the upregulation of genes associated with QS signal production and detection. This results in distinct steady-state concentrations of QS-related molecules in QS-ON and QS-OFF states. Due to the slow decay rates of biomolecules such as proteins, even after removal of the initial stimuli, cells can retain elevated levels of QS-associated biomolecules for extended periods of time. This persistence of biomolecules after the removal of the initial stimuli has the potential to impact the response to future stimuli, indicating a memory of past exposure. This phenomenon, which is a consequence of the carry-over of biomolecules rather than genetic inheritance, is known as "phenotypic" memory. This theoretical study aims to investigate the presence of phenotypic memory in QS and the conditions that influence this memory. Numerical simulations based on ordinary differential equations and analytical modeling were used to study gene expression in response to sudden changes in cell density and extracellular signal concentrations. The model examined the effect of various cellular parameters on the strength of QS memory and the impact on gene regulatory dynamics. The findings revealed that QS memory has a transient effect on the expression of QS-responsive genes. These consequences of QS memory depend strongly on how cell density was perturbed, as well as various cellular parameters, including the Fold Change in the expression of QS-regulated genes, the autoinducer synthesis rate, the autoinducer threshold required for activation, and the cell growth rate.

Effect of Intravitreal Injection of Ranibizumab (Lucentis®) on Corneal Endothelial Cell Density and Morphology

Abstract Background VEGF has an essential role in initiating wound and inflammation-related neovascularization. Several studies have demonstrated that VEGF is an important mediator of blood– retinal-barrier breakdown, leading to fluid leakage below the macula and inducing the development of macular edema and choroidal neovascular membranes. Anti-VEGF agents interfere with its receptor binding, thus inhibiting VEGF’s signal, which results in inhibition of abnormal blood vessel formation and decreasing vascular permeability. Aim of the Work This study aimed is to report early changes in the corneal endothelial cell density (ECD) and morphology in patients undergoing intravitreal anti-VEGF injections (ranibizumab) for treatment of macular edema. Patients and Methods This prospective study included 22 eyes of 22 DME patients indicated for IVI of Ranibizumab, with mean age 57.1±7.4 years, ranging from 42.0 years to 70.0 years. The majority of them were females (77.3%). Results Cell density (endothelial cell count) at baseline (2601.8±220.0) didn’t show statistically significant difference than that at one month (2598.9±198.1) after IVI. Coefficient variant was not significantly changed at month-1 as compared to baseline. Coefficient variant at baseline was 29.8±4.0, ranging from 22.0 to 39.0. and at one month was 31.3±5.4, ranging from 21.0 to 48.0. Hexagonal cells (HEX) were not significantly changed at one month as compared to baseline. Hexagonal cells at baseline were 68.9±3.2, ranging from 63.0 to 75.0 and after one month, it was 67.8±4.1, ranging from 60.0 to 76.0. Pachymetry was not significantly changed at one month as compared to baseline. The corneal thickness (pachymetry) at baseline was 538.8±40.1 ranging from 448.0–587.0 to 447.0–597.0. after one month of IV ranibizumab it was 536.3±42.1 0.183. Conclusion A single IV injection of Ranibizumab in phakic and otherwise normal eyes with DME showed no change in ECD and pachymetry but a statistically non-significant increase in CV and decrease in HEX 1-month after injection.

Integrative transcriptomic and proteomic profiling of the effects of cell confluency on gene expression

In this study we examine the impact of cell confluency on gene expression. We focused on Argonaute (AGO) protein dynamics and associated gene and protein expression in HEK293, A375, and SHSY5Y cell lines. As a consequence of cell confluency, AGO2 protein translocates into the nucleus. Therefore, we generated transcriptomic data using RNA sequencing to compare gene expression in subconfluent versus confluent cells, which highlighted significant alterations in gene regulation patterns directly corresponding to changes in cell density. Our study also encompasses miRNA profiling data obtained through small RNA sequencing, revealing miRNA expressional changes dependent on cellular confluency, as well as cellular localization. Finally, we derived proteomic data from mass spectrometry analyses following AGO1-4 immunoprecipitation, providing a comprehensive view of AGO interactome in both nuclear and cytoplasmic compartments under varying confluency. These datasets offer a detailed exploration of the cellular and molecular dynamics, influenced by cell confluency, presenting a valuable resource for further research in cellular biology, particularly in understanding the basic mechanisms of cell density in cancer cells.

Assessment of the Predictive Ability of Theranostics for Corneal Cross-linking in Treating Keratoconus: A Randomized Clinical Trial

PurposeTo validate the ability of theranostic imaging biomarkers in assessing the propensity of corneal cross-linking (CXL) in flattening the maximum keratometry (Kmax) index. DesignProspective, randomized, multicenter, masked clinical trial (NCT05457647). ParticipantsFifty patients with progressive keratoconus. InterventionParticipants were stratified to undergo epithelium-off (epi-off; 25 eyes) and epithelium-on (epi-on; 25 eyes) CXL protocols using UV-A medical device incorporating theranostic software module. The device used controlled UV-A light both for performing CXL and for estimating the corneal riboflavin concentration (riboflavin score) and assessing treatment effect (theranostic score) in real time. A 0.22% riboflavin formulation was applied onto the cornea for 15 minutes and 20 minutes in epi-off and epi-on protocols respectively. All eyes underwent 9 minutes UV-A irradiance at 10 mW/cm2. Main Outcome MeasuresThe primary outcome measure was validation of the combined use of theranostic imaging biomarkers through measurement of their accuracy (proportion of correctly classified eyes) and precision (positive predictive value) to correctly classify eyes and positively predict a Kmax flattening at 1 year after CXL. Other outcome measures were the change of Kmax, endothelial cell density, uncorrected and corrected distance visual acuity, manifest spherical equivalent refraction, and central corneal thickness one year after CXL. ResultsAccuracy and precision of the combined use of theranostic imaging biomarkers in predicting eyes that had more than 0.1 diopter (D) Kmax flattening at 1 year were 91% and 95% respectively. The Kmax value significantly flattened by a median of -1.3 D (IQR: -2.11, -0.49 D; P < 0.001); both the uncorrected and corrected distance visual acuity improved by a median of -0.1 LogMAR (IQR: -0.3, 0.0 LogMAR; P < 0.001 and IQR: -0.2, 0.0 LogMAR; P < 0.001 respectively). There were no significant changes in endothelial cell density (P = 0.33) and central corneal thickness (P = 0.07) 1 year postoperatively. ConclusionsThe study demonstrated the efficacy of integrating theranostics in a UV-A medical device for the precise and predictive treatment of keratoconus with epi-off and epi-on CXL protocols. The concentration of riboflavin and its UV-A light mediated photo-activation in the cornea are the primary factors determining CXL treatment efficacy.

Corneal thickness and endothelial change after use of ocular hypotensive agents.

Corneal transplantation can restore visual function when visual impairment is caused by a corneal disease. However, this treatment is associated with the scarcity of cornea donors. The suitability of corneal donation from patients with glaucoma using ocular hypotensive agents (OHAs) is controversial. This study aimed to elucidate changes in corneal thickness, corneal endothelial cell density, and corneal endothelial cell hexagonality after OHA use in patients with primary open-angle glaucoma. We retrospectively reviewed the data of 53 glaucoma suspect eyes without OHA use and 106 primary open-angle glaucoma eyes under OHA use. All participants underwent corneal parameter assessment using SP-3000P (Topcon Corp., Tokyo, Japan) at the time of diagnosis and the final visit. The OHA dose and timing of use were recorded. The ocular hypotensive agents score (OHAS) was determined based on the number, formula, frequency, and duration of OHA use. Baseline data showed no significant differences between the two groups with and without OHA use. At the final visit, the OHA-treated group showed significantly lower corneal thickness and corneal endothelial cell density than those of the control group. A weak positive correlation between the OHAS and changes in corneal endothelial cell hexagonality was noted. However, no correlation was observed between the OHAS and changes in corneal thickness or endothelial cell density. In conclusion, patients with glaucoma and using OHAs should undergo the corneal structural properties examinations before donation to ensure the quality of donor cornea.

Local and global changes in cell density induce reorganisation of 3D packing in a proliferating epithelium.

Tissue morphogenesis is intimately linked to the changes in shape and organisation of individual cells. In curved epithelia, cells can intercalate along their own apicobasal axes, adopting a shape named 'scutoid' that allows energy minimization in the tissue. Although several geometric and biophysical factors have been associated with this 3D reorganisation, the dynamic changes underlying scutoid formation in 3D epithelial packing remain poorly understood. Here, we use live imaging of the sea star embryo coupled with deep learning-based segmentation to dissect the relative contributions of cell density, tissue compaction and cell proliferation on epithelial architecture. We find that tissue compaction, which naturally occurs in the embryo, is necessary for the appearance of scutoids. Physical compression experiments identify cell density as the factor promoting scutoid formation at a global level. Finally, the comparison of the developing embryo with computational models indicates that the increase in the proportion of scutoids is directly associated with cell divisions. Our results suggest that apico-basal intercalations appearing immediately after mitosis may help accommodate the new cells within the tissue. We propose that proliferation in a compact epithelium induces 3D cell rearrangements during development.

Assessment of in-situ monitoring and tracking the vertical migration of cyanobacterial blooms using LISST-HAB

Cyanobacterial harmful algal blooms (cyanoHABs) are becoming increasingly common in aquatic ecosystems worldwide. However, their heterogeneous distributions make it difficult to accurately estimate the total algae biomass and forecast the occurrence of surface cyanoHABs by using traditional monitoring methods. Although various optical instruments and remote sensing methods have been employed to monitor the dynamics of cyanoHABs at the water surface (i.e., bloom area, chlorophyll a), there is no effective in-situ methodology to monitor the dynamic change of cell density and integrated biovolume of algae throughout the water column. In this study, we propose a quantitative protocol for simultaneously measurements of multiple indicators (i.e., biovolume concentration, size distribution, cell density, and column-integrated biovolume) of cyanoHABs in water bodies by using the laser in-situ scattering and transmissometry (LISST) instrument. The accuracy of measurements of the biovolume and colony size of algae was evaluated and exceeded 95% when the water bloom was dominated by cyanobacteria. Furthermore, the cell density of cyanobacteria was well estimated based on total biovolume and mean cell volume measured by the instrument. Therefore, this methodology has the potential to be used for broader applications, not only to monitor the spatial and temporal distribution of algal biovolume concentration but also monitor the vertical distribution of cell density, biomass and their relationship with size distribution patterns. This provides new technical means for the monitoring and analysis of algae migration and early warning of the formation of cyanoHABs in lakes and reservoirs.

Methamphetamine and the Synthetic Cathinone 3,4-Methylenedioxypyrovalerone (MDPV) Produce Persistent Effects on Prefrontal and Striatal Microglial Morphology and Neuroimmune Signaling Following Repeated Binge-like Intake in Male and Female Rats.

Psychostimulants alter cellular morphology and activate neuroimmune signaling in a number of brain regions, yet few prior studies have investigated their persistence beyond acute abstinence or following high levels of voluntary drug intake. In this study, we examined the effects of the repeated binge-like self-administration (96 h/week for 3 weeks) of methamphetamine (METH) and 21 days of abstinence in female and male rats on changes in cell density, morphology, and cytokine levels in two addiction-related brain regions-the prefrontal cortex (PFC) and dorsal striatum (DStr). We also examined the effects of similar patterns of intake of the cocaine-like synthetic cathinone derivative 3,4-methylenedioxypyrovalerone (MDPV) or saline as a control. Robust levels of METH and MDPV intake (~500-1000 infusions per 96 h period) were observed in both sexes. We observed no changes in astrocyte or neuron density in either region, but decreases in dendritic spine densities were observed in PFC pyramidal and DStr medium spiny neurons. The microglial cell density was decreased in the PFC of METH self-administering animals, accompanied by evidence of microglial apoptosis. Changes in microglial morphology (e.g., decreased territorial volume and ramification and increased cell soma volume) were also observed, indicative of an inflammatory-like state. Multiplex analyses of PFC and DStr cytokine content revealed elevated levels of various interleukins and chemokines only in METH self-administering animals, with region- and sex-dependent effects. Our findings suggest that voluntary binge-like METH or MDPV intake induces similar cellular perturbations in the brain, but they are divergent neuroimmune responses that persist beyond the initial abstinence phase.

In vivo assessment of human corneal epithelial cells in orthokeratology lens wearers: A pilot study.

Central corneal epithelial thinning associated with midperipheral epithelial thickening has been reported as the main factor contributing to the effectiveness of orthokeratology (ortho-k) in myopia control. Yet, the cellular mechanism governing the regional change in refractive power remains elusive. This study aimed to evaluate the correlation between the regional change in corneal epithelial thickness and cell density in ortho-k wearers. A new human prototype of a polarization-dependent optical coherence microscope was developed to enable noncontact and noninvasive in vivo imaging of corneal epithelial cells in ortho-k wearers with and without their ortho-k lens. The epithelial thickness and cell density were evaluated at the central and midperipheral corneal locations in four ortho-k wearers and four spectacle wearers serving as controls. Polarization-dependent optical coherence microscope achieved in vivo volumetric imaging of all epithelial cell types in ortho-k wearers with and without their lens over a field of view of 0.5 × 0.5 mm2 with an isotropic resolution of ~2.2 mm. The central epithelial thinning and midperipheral epithelial thickening were consistent across all ortho-k wearers. However, the inconsistency in their regional epithelial cell density highlighted a great variability in individual response to ortho-k treatment. There was no strong correlation between epithelial thickness and cell density, especially at the midperipheral cornea, in ortho-k participants. This study constitutes our first step toward uncovering the cellular mechanism underlying the effectiveness of ortho-k in myopia control. Future studies will focus on the longitudinal evaluation of epithelial cells before and during ortho-k treatment to identify factors governing individual response to ortho-k treatment and ultimately inform the dynamics of epithelial cells taking place during the ortho-k treatment.

Enhanced Surgical Safety through Protective Penetrating Keratoplasty: A Retrospective Analysis of 22 Cases.

To assess the surgical safety, postoperative outcomes, and the impact of PPK on corneal endothelial cell density through a retrospective analysis of 22 cases. Settings and Design. A retrospective cross-sectional observational study was executed at Hermanos Ameijeiras Hospital from February 2018 to December 2021, involving 22 patients (22 eyes) who were unresponsive to other medical treatments and had a corrected distance visual acuity of ≤0.1. Patients underwent PPK, with surgical procedures and postoperative care documented. Statistical analysis was performed on qualitative and quantitative variables to evaluate the surgical outcomes and the corneal endothelial cell density changes postoperation. All surgeries demonstrated a significant improvement in postoperative visual acuity (p ≤ 0.001) and recorded a 9.2% decrease in the corneal endothelial cell density at 12 months. Noteworthy complications included one case of intraoperatively discovered haptic dislocation and one postoperative bacterial keratitis. PPK could potentially mitigate perioperative complications, ensure graft clarity, and reduce corneal endothelial cell loss, presenting itself as a viable alternative to traditional PK. Although the results are encouraging, larger-scale studies are essential to validate the benefits and applicability of PPK in broader clinical settings.

Abstract CT134: Safety and immunologic impact of neoadjuvant/adjuvant GM-CSF-secreting allogenic pancreatic tumor cell vaccine (GVAX) combined with cyclophosphamide, pembrolizumab, and macrophage-targeting CSF1R inhibitor IMC-CS4 in pancreatic adenocarcinoma

Abstract Background: Strategies are needed to improve effectiveness of immunotherapy in pancreatic ductal adenocarcinoma (PDA). Our recently published study (NCT02648282) showed that increased M1/M2 ratio and decreased PDL1+ M2-like tumor associated macrophages (TAM) in locally advanced PDA is associated with longer overall survival (OS) following combination of stereotactic body radiotherapy, GVAX and anti-PD1 antibody pembrolizumab (Pem). We hypothesized targeting M2-like macrophages, regulated by the colony stimulating factor-1 (CSF1) pathway, would improve outcomes. In this pilot study we investigated the safety and intratumoral immune response of neoadjuvant/adjuvant triplet immunotherapy: GVAX/cyclophosphamide (CY), Pem, and CSF1 receptor blockade (IMC-CS4) in patients with resectable or borderline resectable PDA. Methods: After completing chemotherapy and radiation patients received two 3-week cycles of neoadjuvant triplet immunotherapy. Following surgery, patients received 4 additional cycles followed by booster Pem every 3 weeks and GVAX/CY every 6 months, for one year. Co-primary endpoints were safety, assessed by NCI Common Terminology Criteria for Adverse Events (AEs), and immune response (increased CD8+ T-cell density in surgical specimen as compared to pre-treatment core). Secondary endpoints included disease free survival (DFS), OS, surgical resectability, and pathologic response (PR). Results: Nine patients were enrolled. Two immune related grade 3/4 AEs (diarrhea and rash) were reported. Median DFS was 12.6 months (95% CI: 7.3, NR) and median OS was 20.4 months (95% CI: 17.5, NR). All patients underwent surgery with major PR rate of 78% (defined as &amp;lt;10% residual viable tumor), including one complete PR. Comparing paired biopsies, the study met its primary immunologic endpoint showing 6/8 patients (75%) had a &amp;gt;80% increase of CD8+ T cells; the increase was at least 1.8 times the baseline median absolute deviation following triple therapy (p=0.012). There was a significant increase of Granzyme B+ CD8+ T cells among all patients (p=0.045). No significant change in myeloid cell density, including TAM, was observed, suggesting IMC-CS4 did not deplete macrophages, but potentially reprogrammed them. The 4 patients with greatest drop in CD66b+ granulocyte levels had major PR (grade 0 or 1). Conclusion: The triplet was safe and significantly increased activated CD8+ T cells suggesting the combination can induce an effector T cell response. Decreased granulocyte levels correlated with PR. Citation Format: Arielle Urman, Yingjun Ding, Hao Wang, Richard A. Burkhart, Jin He, Elizabeth M. Jaffee, Amol K. Narang, Dung Le, Daniel A. Laheru, Lei Zheng, Ana De Jesus-Acosta. Safety and immunologic impact of neoadjuvant/adjuvant GM-CSF-secreting allogenic pancreatic tumor cell vaccine (GVAX) combined with cyclophosphamide, pembrolizumab, and macrophage-targeting CSF1R inhibitor IMC-CS4 in pancreatic adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT134.

Repositioned versus exchanged flanged intraocular lens fixation for intraocular lens dislocation

This study aimed to compare the outcomes of flanged intraocular lens (IOL) fixation with new IOL exchange after dislocated IOL removal and repositioned dislocated IOL in patients with IOL dislocation. Eighty-nine eyes that underwent flanged IOL fixation were retrospectively included, with 51 eyes in the exchanged IOL group and 38 eyes in the repositioned IOL group. In both groups, best-corrected visual acuity (BCVA) improved at 1, 3, 6, and 12 months postoperatively and did not differ between the two groups at any of these time points. However, at 1 week postoperatively, BCVA in the repositioned IOL group improved compared with baseline, whereas that in the exchanged IOL group did not. Moreover, there were lesser changes in the corneal endothelial cell density (ECD) and corneal astigmatism in the repositioned IOL group than in the exchanged IOL group. The IOL positions, including IOL tilt and IOL decentration, were not different between the groups. Flanged IOL fixation with new IOL exchange and with repositioned dislocated IOL for patients with IOL dislocation had similar visual outcomes and IOL position. However, the latter had a smaller corneal ECD decrease and astigmatic change. This technique was effective in treating IOL dislocation while minimizing corneal injury.

Clinical Evaluation of Corneal Endothelial Parameters following Laser Refractive Surgery in Myopic Eyes: A Review.

Background: The aim of this review was to investigate the influence of various laser refractive surgery methods on the corneal endothelium in myopic patients. The role of the corneal endothelium in laser refractive surgery (LRS) is currently being addressed in the assessment of postoperative corneal edema risk. Methods: Changes in corneal endothelial cell density and morphology after LRS were evaluated based on a systematic review of current studies. The results of a literature search in the PubMed, Science Direct, Google Scholar, and the Web of Science databases, as well as a manual search, were selected for the final review according to the PRISMA 2020 flow diagram. Results: We included 24 prospective clinical trials in the review: surface ablation (twelve), LASIK and FemtoLASIK (two), femtosecond lenticule extraction (two), and comparable studies (eight). Endothelial cell density was determined by specular or in vivo confocal microscopy. In most studies, no statistically significant differences were found between preoperative and postoperative endothelial parameters. In nine studies, the changes were statistically significant, but no vision-threatening complications occurred, and no serious corneal complications developed in any eyes during the follow-up period. Conclusions: Based on collected data, laser keratorefractive surgery appears not to exert a significant effect on the corneal endothelium.

Production and release of 2-MIB in Pseudanabaena: Effects of growth phases on cell characteristics and 2-MIB yield

2-methylisoborneol (2-MIB), a secondary metabolite produced by cyanobacteria, often causes a musty odour in water, threatening the safety of drinking water supplies. This study investigated the effects of the growth phases on the production of 2-MIB by Pseudanabaena. The effects of cell characteristics on the production and release of 2-MIB were also explored. The total 2-MIB concentration increased during the exponential phase and decreased during the declining phase, which was consistent with the changes in cell density. However, the total 2-MIB yield (1.12–1.27 fg cell−1) of Pseudanabaena did not significantly differ throughout the growth cycle (p > 0.05). Meanwhile, the extracellular 2-MIB yield increased significantly from 0.31 fg cell−1 in the exponential phase to 0.76 fg cell−1 in the declining phase (p < 0.05), and the corresponding proportion of extracellular 2-MIB improved from 25.13% to 59.16% (p < 0.05). The surge in extracellular 2-MIB during the declining phase could be attributed to the breaking of the Pseudanabaena filament, as indicated by the decrease in Dmean during cell ageing. The findings of this study contribute to a more inclusive comprehension and management of musty odour issues resulting from cyanobacteria in the water supply.