Cell Density Research Articles

Insights into the response mechanisms of Tetradesmus obliquus to aged polylactic acid and tetracycline exposure via transcriptome analysis and physiological evaluations

Microplastics (MPs) and antibiotics, identified as emerging pollutants, are extensively prevalent in aquatic environments and display prolonged durability. Unlike conventional plastics, biodegradable plastics are more susceptible to decomposition in the environment, resulting in the generation of microplastics and posing potential risks to the aquatic ecosystems. In this study, we assessed growth inhibition, chlorophyll a content, malondialdehyde content (MDA), and antioxidant enzyme activities. These measurements were integrated with transcriptome analysis to explore the response mechanisms of virgin and aged polylactic acid (vPLA and aPLA) and tetracycline (TC) following 14-day exposure to Tetradesmus obliquus, either individually or in combination. The findings indicated that exposure to vPLA did not significantly impact the growth of T. obliquus. Conversely, aPLA demonstrated growth-promoting effects on T. obliquus, particularly in the latter incubation stages. Moreover, a 14-day exposure significantly increased the chlorophyll a content and the activities of superoxide dismutase (SOD), catalase glutathione (CAT) and glutathione S-transferase (GST) within the algal cells. Apart from 1 mg L−1, the TC concentrations of 2.5, 5.0, and 10 mg L−1 exhibited significant toxic effects on T. obliquus, including growth inhibition, decreased chlorophyll a content, elevated activities of SOD, CAT, and GST, and increased MDA levels. Exposure to a combination of 300 mg L−1 aPLA and 5.0 mg L−1 TC, compared to solely 5 mg L−1 TC, demonstrated a notable reduction in TC toxicity to T. obliquus in the presence of aPLA. This was indicated by elevated algal cell density and chlorophyll a content, as well as a decrease in MDA content. Transcriptome analysis indicated an enrichment of differentially expressed genes (DEGs) in pathways linked to porphyrin metabolism, photosynthesis, carbon fixation, and metabolism within the aPLA + TC combined exposure. The study aid in expanding our knowledge of the potential ecological risks posed by biodegradable plastics and accompanying pollutants in aquatic environments.

Detecting cell types and densities in the tumor microenvironment improves prognostic risk assessment for breast cancer.

A comprehensive evaluation of the relationship between the densities of various cell types in the breast cancer tumor microenvironment and patient prognosis is currently lacking. Additionally, the absence of a large patch-level whole slide imaging (WSI) dataset of breast cancer with annotated cell types hinders the ability of artificial intelligence to evaluate cell density in breast cancer WSI. We first employed Lasso-Cox regression to build a breast cancer prognosis assessment model based on cell density in a population study. Pathology experts manually annotated a dataset containing over 70,000 patches and used transfer learning based on ResNet152 to develop an artificial intelligence model for identifying different cell types in these patches. The results showed that significant prognostic differences were observed among breast cancer patients stratified by cell density score (P = 0.0018), with the cell density score identified as an independent prognostic factor for breast cancer patients (P < 0.05). In the validation cohort, the predictive performance for overall survival (OS) was satisfactory, with area under the curve (AUC) values of 0.893 (OS) at 1-year, 0.823 (OS) at 3-year, and 0.861 (OS) at 5-year intervals. We trained a robust model based on ResNet152, achieving over 99% classification accuracy for different cell types in patches. These achievements offer new public resources and tools for personalized treatment and prognosis assessment.

A novel bioprinted microtumour model for studying acute-leukemia-cell- contaminated in ovarian tissue

Microtumor models, combining cancer and stromal cells within 3D hydrogels, are vital for testing anticancer therapies. Bioprinting hydrogel scaffolds allows tailored in vitro models. We created a 3D microtumor model using a bioprinter, with varying ratios of ovarian stromal cells and leukemia cells (HL-60). PEGylated fibrinogen and alginate hydrogel were used. Cell dynamics and proliferation were assessed via immunofluorescence staining. Microtumors with different HL-60 ratios (1:1, 1:10, 1:100) were cultured for 5 days. Results showed tumor development modulation by cell ratios and culture time. A significant cell density increase occurred in 1:1 ratio microtumors, indicating rapid cancer cell proliferation. No HL-60 cells were found in 1:100 ratio microtumors by day 5. The 1:10 ratio closely mimicked leukemia invasion in ovarian tissue, showing detectable cancer cells by days 3 and 5 without altering total cell density dynamics significantly. This bioprinted leukemia microtumor model offers better physiological relevance than 2D assays, promising applications in cellular analysis and drug screening.

Phenotypic variability of Montastraea cavernosa and Porites astreoides along a depth gradient from shallow to mesophotic reefs in the Cayman Islands

Phenotypic variability is the ability of the same species to express different phenotypes under different environmental conditions. Several coral species that exist along a broad depth distribution have been shown to differ in skeletal morphology and nutrient acquisition at different depths, which has been attributed to variability in response to differing levels of light availability. This study examined the phenotypic variability of two common depth generalist corals, Montastraea cavernosa and Porites astreoides, along a gradient from 10 to 50 m in the Cayman Islands, by examining changes in skeletal morphology, photophysiology, symbiont cell density, and chlorophyll concentration. Skeletal features of M. cavernosa were found to increase in size from 10 to 30 m, but returned to smaller sizes from 30 to 50 m, while P. astreoides skeletal features continued to increase in size from 10 to 40 m. No differences were observed in either symbiont density or chlorophyll concentration across depths for either species. However, all photophysiological parameters exhibited significant depth-dependent variations in both species, revealing adaptive strategies to different light environments. These results suggest that both species have high variability in response to depth. Patterns of skeletal morphology and photophysiology, however, suggest that M. cavernosa may be more variable in regulating photosynthetic efficiency compared to P. astreoides, which likely facilitates the broader depth distribution of this species.

UiO‐66‐NH2 anchored sulfonated polyphenylene sulfide proton exchange membrane with high proton conductivity and low methanol permeability

AbstractThis study loaded different amounts of UiO‐66‐NH2 onto sulfonated polyphenylene sulfide fibers (SPPS), successfully loading UiO‐66‐NH2 on SPPS fibers confirmed by energy dispersive x‐ray spectroscopy, x‐ray diffraction, and Fourier transform infrared spectroscopy. The UiO‐66‐NH2‐X@SFM were then impregnated in Nafion solution to obtain UiO‐66‐NH2‐X@SFM/Nafion composite membranes. The thermal stability, dimensional stability, mechanical properties, proton conductivity, methanol resistance, and other properties of the UiO‐66‐NH2‐X@SFM/Nafion composite membranes were subsequently comprehensively characterized. The proton conductivity of the UiO‐66‐NH2‐8@SFM/Nafion composite membrane at 80°C and 100% RH was measured to be 0.286 S cm−1, which exhibited a significant enhancement of 1.75 times compared with the original membrane. Simultaneously, the composite membrane demonstrated a remarkable reduction in methanol permeation rate. At 40°C and 100% RH, the UiO‐66‐NH2‐8@SFM/Nafion composite membrane has a selectivity of up to 20.7 × 104 S s cm−3 and a single cell density of 88.3 m W cm−2 at 60°C and 100% RH.

Safety and prognosis of phacoemulsification using active sentry and active fluidics with different IOP settings - a randomized, controlled study

PurposeTo explore the impact of different intraoperative intraocular pressure (IOP) settings on the safety and prognosis in phacoemulsification.MethodsAge related cataract patients who met the inclusion criteria and underwent phacoemulsification by using active sentry handpiece and active fluidics system. According to different intraoperative IOP settings during surgery, they were randomly divided into two groups: the 20mmHg group and the 60mmHg group. The best corrected visual acuity (BCVA), cumulative dissipated energy (CDE), total U/S time, active surge mitigation (ASM), estimated fluid usage (EFU) as well as the changes in corneal thickness (CT), corneal epithelial layer thickness (CELT) and endothelial cell density (ECD) were collected. The post-operative follow-up was only 1 day.ResultsA total of 110 cases (110 eyes) were included in the study. There were 55 eyes in each group. There was no statistically significant difference in postoperative BCVA (p = 0.839). The CDE, total U/S time and EFU during surgery were (5.22 ± 3.31), (30.60 ± 15.06), (45.07 ± 12.68) and (4.70 ± 2.83), (27.39 ± 13.75), (42.38 ± 11.93) in the 20mmHg group and 60mmHg group (p = 0.381, 0.246, 0.254). The ASM during surgery in the 20mmHg group and 60mmHg group were (0.95 ± 2.77) and (7.24 ± 6.34), respectively. The 20mmHg group showed a significant decrease in ASM (p < 0.001). There was no statistically significant difference in the changes in CT, CELT and ECD before and after surgery between the two groups (p = 0.913, 0.825, 0.624). Both groups did not experience any intraoperative complications, such as posterior capsule rupture.ConclusionA lower IOP setting of 20 mmHg can significantly reduce the occurrence of intraoperative surges during phacoemulsification. And there was no increase in rate of complications.Trial RegistrationThe trial registration number is ChiCTR2100050240. The registered date is August 24th, 2021.

The predictive value of anterior segment optical coherence tomography for postoperative corneal edema in cataract patients.

To evaluate the predictive value of anterior segment optical coherence tomography (AS-OCT) for postoperative corneal edema in cataract patients. A retrospective cohort study of 144 cataract patients from December 2020 to December 2021 was conducted. Patients were divided into edema eyes (84 cases) and observation (60 cases) group based on postoperative corneal edema occurrence. Relevant indicators were compared between groups. Logistic regression identified risk factors for postoperative corneal edema. Receiver operating characteristic curves evaluated the predictive value. The edema eyes group had significantly higher postoperative central corneal thickness (CCT) and lower postoperative endothelial cell density (ECD) than the observation group (P < 0.05). The edema eyes group also had significantly lower preoperative ECD, anterior chamber depth (ACD), anterior chamber angle (ACA), and preoperative lens position (LP) than the observation group (P < 0.05). Logistic regression analysis showed that preoperative ECD, ACD, ACA, and LP were independent risk factors for postoperative corneal edema (P < 0.05), all of which also showed good predictive value for postoperative corneal edema, with areas under the curve (AUCs) of 0.854, 0.812, 0.791, and 0.778, respectively, under the ROC curve analysis. AS-OCT can provide useful information for predicting postoperative corneal edema in cataract patients. Preoperative ECD, preoperative ACD, preoperative ACA, and preoperative LP are important parameters that can be measured by AS-OCT and used as risk factors for postoperative corneal edema.

Significant Reduction of Radiation-Induced Death in Mice Treated with PrC-210 and G-CSF after Irradiation.

The search for single or combined radiation countermeasures that mitigate the development of Acute Radiation Syndrome (ARS) after radiation exposure remains a prominent goal of the U.S. government. This study was undertaken to determine whether PrC-210 and G-CSF, when administered 24-48 h postirradiation, would confer an additive or synergistic survival benefit and mitigate ARS in mice that had received an otherwise 96% lethal radiation dose. Our results show that optimum systemic doses of PrC-210 and G-CSF, when administered 24 h or later after a 96% lethal dose of whole-body irradiation, conferred: 1. strong individual survival benefits (PrC-210 44%, P = 0.003), (G-CSF 48%, P = 0.0002), 2. a profound combined 85% survival benefit (P < 0.0001) when administered together, and on day 14 postirradiation, 3. peripheral white blood cell/lymphocyte counts equal to unirradiated controls, 4. dense bone marrow cell density (>65% of unirradiated controls), 5. jejunal villi density that equaled 90% of unirradiated controls, and 6. spleen weights that equaled 93% of unirradiated controls. Our results show that PrC-210 and G-CSF given together 24 h after irradiation confer strong additive efficacy by protecting the immune system, and enabling recovery of the bone marrow, and they work synergistically to enable recovery of peripheral white blood cells in circulating blood.

Active Magnesium Boride/Alginate Hydrogels Rejuvenate Senescent Cells.

The clearance of senescent cells may be detrimental to low cell density diseases, such as intervertebral disc degeneration (IVDD), and rejuvenating these cells presents a formidable obstacle. In this study, we investigate a mild-alkalization strategy employing magnesium boride-alginate (MB-ALG) hydrogels to rejuvenate senescent cells associated with age-related diseases. MB-ALG hydrogels proficiently ensnare senescent cells owing to their surface roughness. The hydrolysis of MB-ALG hydrogels liberates hydroxide ions (OH-), effecting a transition from an acidic microenvironment (pH ∼ 6.2) to a mildly alkaline state (pH ∼ 8.0), thereby fostering senescent cell proliferation via activation of the PI3K/Akt/mTOR pathway. Additionally, H2 aids in ROS clearance, which reduces cellular oxidative stress. And, Mg2+ rejuvenates senescent cells by inhibiting Ca2+ influx and fine-tuning the sirt1-p53 signaling pathways. Both in vitro and in vivo experiments conducted on rat intervertebral discs corroborate the sustained antisenescence and rejuvenation properties of MB-ALG hydrogels, with effects persisting for up to 12 weeks postoperation. These discoveries elucidate the role of mild-alkalization in dictating cellular destiny and provide key insights for addressing age-related diseases.

Spermatogenesis in mouse testicular organoids with testis-specific architecture, improved germ cell survival and testosterone production

This study presents a biphasic approach to overcome the limitations of current testicular organoid (TO) cultures, including histological heterogeneity, germ cell loss and absence of spermatogenesis. Agarose microwells were utilized to create TOs from prepubertal C57BL/6 J testicular cells. First emphasis was on improving germ cell survival during the initial 2-week reorganization phase by comparingα-MEM + 10% knockout serum replacement (KSR) medium, known to support TO generation in mice, to three optimized media (1-3). Cell densities and culture dynamics were also tested to recreate histological resemblance to testes. After optimizing germ cell survival and cell organization, the effect of growth factors and immunomodulation through CD45+immune cell depletion or dexamethasone (DEX) supplementation were assessed for enhancing spermatogenesis during the subsequent differentiation phase. Testicular cells self-reorganized into organoids resembling the testicular anatomical unit, characterized by one tubule-like structure surrounded by interstitium. Media 1-3 proved superior for organoid growth during the reorganization phase, with TOs in medium 3 exhibiting germ cell numbers (7.4% ± 4.8%) comparable to controls (9.3% ± 5.3%). Additionally, 37% ± 30% demonstrated organized histology from 32 × 103cells under static conditions. Switching toα-MEM + 10% KSR during the differentiation phase increased formation efficiency to 85 ± 7%, along with elevated germ cell numbers, testosterone production (3.1 ± 0.9 ng ml-1) and generation ofγ-H2AX+spermatid-like cells (steps 8-11, 1.2% ± 2.2% of the total). Adding differentiation factors to theα-MEM increased spermatid-like cell numbers to 2.9% ± 5.9%, confirmed through positive staining for CREM, transition protein 1, and peanut agglutinin. Although, these remained diploid with irregular nuclear maturation. DEX supplementation had no additional effect, and immune cell depletion adversely impacted TO formation. The manipulability of TOs offers advantages in studying male infertility and exploring therapies, with scalability enabling high-throughput chemical screening and reducing animal usage in reproductive toxicity and drug discovery studies.

Expression of Matrix Metalloproteinase-3 in the Aqueous Humor of Patients with Posner-Schlossman Syndrome

Purpose To explore the expression of matrix metalloproteinase-3 (MMP-3) in the aqueous humor of patients with Posner-Schlossman syndrome (PSS), and the association between MMP-3 and PSS. Methods Peripheral blood and aqueous humor were routinely collected from 29 patients with PSS (PSS group) and 30 patients with age-related-cataract (ARC) (control group). The content of MMP-3 in serum and aqueous humor was measured by immunoturbidimetry. The correlation between MMP-3 and ophthalmic examination results were verified by Spearman’s correlation analysis. Results The MMP-3 level in the aqueous humor of the PSS group was (25.86 ± 13.4)ng/ml, significantly higher than that in the control group (3.9 ± 2.7)ng/ml(p < 0.001), while there was no significant difference in serum MMP-3 level between the two groups (p = 0.125). The endothelial cell density (ECD) in the aqueous humor of the PSS group was (2078 ± 440) cell/mm2, intraocular pressure (IOP) in the aqueous humor of the PSS group was (33 ± 12) mmHg. The correlation analysis of aqueous humor MMP-3 and various ophthalmic examination results showed that aqueous humor MMP-3 had a moderate correlation with IOP and the difference in ECD between the affected eye and the fellow eye. Conclusions MMP-3 level is elevated in the aqueous humor of PSS patients, and it may play an important role in the pathogenesis of PSS.

Enhancing protein productivities in CHO cells through adenosine uptake modulation – Novel insights into cellular growth and productivity regulation

Maximizing production potential of recombinant proteins such as monoclonal antibodies (mAbs) in Chinese Hamster Ovary (CHO) cells is a key enabler of reducing cost of goods of biologics. In this study, we explored various strategies to utilize adenosine mediated effects in biologics manufacturing processes. Results show that supplementation of adenosine increases specific productivity by up to two-fold while also arresting cell growth. Introducing adenosine in intensified perfusion processes in a biphasic manner significantly enhanced overall productivity. Interestingly, adenosine effect was observed to be dependent on the cell growth state. Using specific receptor antagonists and inhibitors, we identified that ENTs (primarily Slc29a1) mediate the uptake of adenosine in CHO cell cultures. Transcriptomics data showed an inverse correlation between Slc29a1 expression levels and peak viable cell densities. Data suggests that in fed-batch cultures, adenosine can be produced extracellularly. Blocking Slc29a1 using ENT inhibitors such as DZD and DP alone or in combination with CD73 inhibitor, PSB12379, resulted in a twofold increase in peak viable cell densities as well as productivities in fed batch – a novel strategy that can be applied to biologics manufacturing processes. This is the first study that suggests that adenosine production/accumulation in CHO cell cultures can potentially regulate the transition of CHO cells from exponential to stationary phase. We also demonstrate strategies to leverage this regulatory mechanism to maximize the productivity potential of biologics manufacturing processes.

Fabrication of smart nanogel based on carrageenan and green coffee extract as a long-term antifouling agent to improve biofilm prevention in food production

This study investigates the extract of the bioactive compounds from green coffee extract (GCE) and the loading of two different concentrations of GCE (1% and 2%) onto carrageenan nanogels (CAR NGs) to compare their antibacterial and antibiofilm effects with unloaded nanogels (NGs). The bioactive compounds of GCE were characterized using GC–MS analysis. The GCE1 and GCE2 were successfully deposited onto the surface of CAR NGs. The antibacterial and antibiofilm potential of prepared NGs were conducted against some foodborne pathogens (E. coli O157, Salmonella enterica, Staphylococcus aureus, and Listeria monocytogenes). The results of GC–MS analysis indicated that there were identified 16 bioactive compounds in GCE, including caffeine (36.27%), Dodemorph (9.04%), and D-Glycero-d-ido-heptose (2.44%), contributing to its antimicrobial properties. The antibacterial coatings demonstrated a notable antimicrobial effect, showing zone of inhibition (ZOI) diameters of up to 37 mm for GCE2 loaded CAR NGs. The minimum inhibitory concentration (MIC) values for GCE2 loaded CAR NGs were 80 ppm for E. coli O157, and 120 ppm for S. enterica, S. aureus, and L. monocytogenes, achieving complete bacterial inactivation within 10–15 min of exposure. Both GCE1 and GCE2 loaded CAR NGs significantly reduced biofilm cell densities on stainless steel (SS) materials for E. coli O157, S. enterica, S. aureus, and L. monocytogenes, with reductions ranging from 60% to 95%. Specifically, biofilm densities were reduced by up to 95% for E. coli O157, 89% for S. enterica, 85% for S. aureus, and 80% for L. monocytogenes. Results of the toxicity evaluation indicated that the NGs were non-toxic and biocompatible, with predicted EC50 values proved their biocompatibility and safety. These results recommended that GCE loaded CAR NGs are promising as natural antimicrobial agents for enhancing food safety and extending shelf life. Further, the study concluded that incorporating GCE into CAR NGs is an effective strategy for developing sustainable antimicrobial coatings for the food industry and manufacturing.

Impact of Donor, Host, and Surgical Parameters on High Endothelial Cell Density More Than 5 Years after Penetrating Keratoplasty.

To investigate the correlation between postoperative endothelial cell loss (ECL) and donor, host, and surgical parameters, and to assess the clinical impact of maintaining a high endothelial cell density (ECD) of ≥ 1500 cells/mm2 5 years after penetrating keratoplasty (PKP). This retrospective cohort study included 216 eyes with 5 years of follow-up, of which 94 had annual visits, and who underwent normal-risk elective PKP for noninfectious indications by one corneal microsurgeon (B. S.) between 2009 and 2016. Among the 216 eyes, ECL (39.1%) over 5 years postoperative exhibited weak positive correlations with storage solution time (p = 0.024) and postmortem time (p = 0.028), and moderately positively correlations with the preoperative ECD (p < 0.001). The 5-year postoperative ECL differed significantly between in domo-prepared (36.8%) and ex domo donor corneas (46.3%; p = 0.001). In the 94 eyes, no significant differences were found between the two groups for central pupil pachymetry (CCT) and BCVA (p > 0.074). However, CCT increased significantly between 1 and 4 years (p = 0.034) and 1 and 5 years postoperatively (p = 0.012), respectively. BCVA improved significantly at 1 year postoperatively and continued to improve until 2 years postoperatively (p < 0.001). The Lions corneal bank Saar-Lor-Lux achieved a significantly reduced ECL (36.8%) over 5 years compared to ex domo donor corneas (46.3%). A weak positive correlation was found between ECL with the storage solution time and the postmortem time, as well as a moderate positive correlation with the preoperative ECD. Although CCT increased significantly over 5 years, BCVA improved significantly from the first to the second postoperative year and remained stable thereafter.

Numerical investigations on electrolytic performance of proton exchange membrane electrolysis cell with parallel flow field

In this study, a three-dimensional simulation model of PEMEC with parallel flow field was established, the heat and mass transfer, electrochemical reactions were described to investigate the electrolytic performance. The distribution characteristics of temperature, liquid water and gas products in the electrolysis cell were obtained. The results show that during the operation of PEMEC system, the liquid water content decreases sequentially along the flow channel direction, while there is a significant temperature gradient in the flow direction of the electrolysis cell. The increase of the initial temperature can increase the current density of the electrolytic cell when the temperature increases from 60 ℃ to 80 ℃, the current density can be increases by 7.86 %, but it can also lead to the dehydration of PEMEC, resulting in thermal degradation of PEMEC, and further the reactivity and proton transfer performance were decreased. It is also found that the increase of liquid water inlet flow not only increased the reactant concentration, accelerated the chemical reaction rate, but also took away more heat generated by ohmic polarization as a coolant, which was conducive to the improvement of electrolytic performance. As the inlet flow rate increased from 1 mL/s to 3 mL/s, the current density at 2.4 V and 1.4 V can be increased by 3.66 % and 1.77 %, respectively, and the maximum temperature can be reduced by 7.4 K.

Compositional change of bacterial communities in oil-polluted seawater amid varying degrees of nanoplankton bacterivory

Petroleum hydrocarbons are being released into the marine environment continuously. They will undergo weathering and may eventually be biodegraded by bacteria and other microbes. While nanoplankton (2–20 μm) are the major consumers of marine bacteria, their effect on the process of biodegradation of oil hydrocarbons is still debated. A 14-day microcosm experiment was conducted to investigate the effects of crude oil hydrocarbons on nanoplankton bacterivory and bacterial community in coastal waters. The coefficients of population growth (0.56–1.80 d−1 for all treatments considered) and grazing mortality (0.38–1.65 d−1 for all treatment considered) of bacteria estimated with the dilution method did not differ among the treatments of control (Ctrl), low dose chemically dispersed oil (LDOil, 2 μL L−1 of crude oil), and high dose chemically dispersed oil (HDOil, 8 μL L−1 of crude oil). Bacterial abundance ranged between 0.21–0.86 × 106 cells mL−1 on average for all treatments. The lack of drastic increases in the cell density of bacterial cells in the oil-loaded treatments was observed throughout the experiment period. Sequencing analysis of the 16S rRNA gene revealed the progressive changes in the community compositions of bacteria in all treatments. The relatively high abundance of oil-degrading bacteria, including Cycloclasticus and Alcanivorax on Days 3–14 of the experiment reflected the presence of biodegradation of oil in the LDOil and HDOil treatments. Throughout the 14 days, the community composition of bacteria in the LDOil and HDOil treatments became more similar and they both differed from that in the Ctrl treatment. This study concluded that, in oil-polluted seawater, the changes in the bacterial community composition were mainly resulting from the addition of chemically dispersed crude oil.

Ten-Year Clinical Outcomes of V4c Implantable Collamer Lens Implantation: Longitudinal Analysis of Visual Acuity, Endothelial Cell Density, and Vault Dynamics

This study aimed to evaluate the long-term clinical outcomes of V4c Implantable Collamer Lens (ICL) implantation for myopia correction over a 10-year period. Retrospective interventional case series METHODS: This retrospective interventional study involved 103 myopic eyes from 54 patients. Visual outcomes, including visual acuity and manifest refraction were assessed. Annual measurements of endothelial cell density (ECD), vault, anterior chamber depth, and intraocular pressure were analyzed using mixed-effects models. At the 10-year follow-up, the mean Logarithm of the Minimum Angle of Resolution (logMAR) uncorrected distance visual acuity and corrected distance visual acuity (CDVA) were -0.037 ± 0.052 and -0.041 ± 0.046, respectively. The mean efficacy and safety indices were 1.11 ± 0.13 and 1.12 ± 0.12. A loss of one line of CDVA occurred in 2% of cases, while 59% gained one or more lines. Postoperative manifest refraction spherical equivalent (MRSE) was within ±0.5 diopters (D) and ±1.0 D in 74% and 99% of cases, respectively, with a mean MRSE of -0.19 ± 0.41 D. Data for ECD and vault fit an exponential decay model, demonstrating a continuous yet gradually slowing decrease over time. Decay rates were 0.012 ± 0.001 per year for ECD and 0.056 ± 0.003 per year for the vault. Notably, ECD lower outliers had significantly lower preoperative ECD values. There were no instances of cataract formation, angle closure, glaucoma, or ICL removal throughout the study. The 10-year outcomes of V4c ICL implantation demonstrate effective and safe visual results. Both ECD and vault exhibit exponential decay patterns over the decade. Patients with lower preoperative ECD values require careful post-implantation monitoring.

Corneal Endothelial Cell Density and Morphology in Healthy Libyan Eyes

Background Endothelial cell density and morphology vary across ethnic groups. Aim This article investigates the corneal endothelial cell density and morphology in healthy Libyan eyes. Methods A 3-month cross-sectional observational study was conducted at Benghazi Teaching Eye Hospital, involving 198 eyes of 100 healthy Libyan participants. The noncontact Topcon specular microscope (SP-1P model) was used to assess the following parameters: the mean central corneal thickness (CCT), the mean cell density (MCD), the mean coefficient of variation (CV%), and the mean percentage of the hexagonal cell (Hex [%]). The variables were analyzed in relation to age and gender using the Statistical Package for the Social Sciences (SPSS version 25.0) Results The mean age of participants in this study was 47.4 ± 13.8 years (range 21–75 years). The mean CCT was 516.45 ± 43.04 μm, the MCD was 2664.30 ± 371.26 cells/mm2, the mean CV% was 32.3% ± 3.7, and the mean Hex (%) was 52.8% ± 9.6. There was no statistical difference in the age, CCT, and MCD across genders. Whereas CV (%) and Hex (%) showed significant gender differences (p &lt; 0.01 for both). There was a significant negative weak correlation between CCT (r = –0.10) and age, as well as a significant negative moderate correlation between MCD and Hex (%) with age (r = –0.36 and r = –0.31, respectively). CV% exhibited a significant, moderately positive association with age (r = 0.35). The higher endothelial cell loss rate of 8.4% was in the third decade of life whereas other age groups ranged between 1.1 and 2.7%. Conclusion The normative data for the endothelium of Libyan eyes are reported, which can be used as a baseline for future studies.

Follicular Immune Landscaping Reveals a Distinct Profile of FOXP3hiCD4hi T Cells in Treated Compared to Untreated HIV.

Follicular helper CD4hi T cells (TFH) are a major cellular pool for the maintenance of the HIV reservoir. Therefore, the delineation of the follicular (F)/germinal center (GC) immune landscape will significantly advance our understanding of HIV pathogenesis. We have applied multiplex confocal imaging, in combination with the relevant computational tools, to investigate F/GC in situ immune dynamics in viremic (vir-HIV), antiretroviral-treated (cART HIV) People Living With HIV (PLWH) and compare them to reactive, non-infected controls. Lymph nodes (LNs) from viremic and cART PLWH could be further grouped based on their TFH cell densities in high-TFH and low-TFH subgroups. These subgroups were also characterized by different in situ distributions of PD1hi TFH cells. Furthermore, a significant accumulation of follicular FOXP3hiCD4hi T cells, which were characterized by a low scattering in situ distribution profile and strongly correlated with the cell density of CD8hi T cells, was found in the cART-HIV low-TFH group. An inverse correlation between plasma viral load and LN GrzBhiCD8hi T and CD16hiCD15lo cells was found. Our data reveal the complex GC immune landscaping in HIV infection and suggest that follicular FOXP3hiCD4hi T cells could be negative regulators of TFH cell prevalence in cART-HIV.

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.