Tomographic, Biomechanical, and In Vivo Confocal Microscopic Changes in Cornea in Chronic Gout Disease.

PURPOSE: This study aimed to evaluate the tomographic, biomechanical, and in vivo confocal microscopic (IVCM) effects of chronic gout disease on human cornea. METHODS: This prospective study included 60 eyes of 30 participants with chronic gout disease and 60 eyes of 30 healthy controls. Corneal thickness, keratometry (K) readings, and corneal aberrations were measured with Sirius 3 D corneal tomography system (Sirius, CSO, Italy). Corneal biomechanical properties (corneal hysteresis [CH], corneal resistance factor [CRF], and intraocular pressure [IOP] parameters) were assessed with an ocular response analyzer (ORA, Reichert Ophthalmic Instruments). The number and morphology of corneal endothelial cells and the number of basal epithelial cells were evaluated with IVCM (Confoscan 4.0). Tear breakup time (TBUT) was also evaluated. RESULTS: The mean diagnosis time of the patients with gout was 91.2 ± 69.6 months (12-300 month). Among corneal tomography measurements, K readings were similar between the two groups, while total and higher-order aberrations(coma, trefoil,s pherical, and quadrafoil) were significantly higher in the gout group. In the evaluation of biomechanical measurements, the CH value was significantly lower and the corneal-compensated IOP value was significantly higher in the gout group (p = 0.02, p = 0.01, respectively). The two groups did not significantly differ regarding the CRF or Goldmann IOP (p = 0.61, p = 0.15, respectively). Among the IVCM parameters, the number of corneal basal epithelial cells and the percentage of corneal endothelial hexagonality were significantly lower in the gout group, but no significant difference was detected in terms of the number of endothelial cells or polymegathism (p = 0.02, p < 0.001, p = 0.18, p = 0.59, respectively). While TBUT was significantly lower in the gout group (p < 0.001). CONCLUSION: This study showed that chronic gout disease increases the corneal aberrations and decreases the basal epithelial cell count, hexagonality ratio of endothelial cell and corneal biomechanics.

Non-invasive techniques in the clinical diagnosis of cutaneous lymphomas.

In clinical practice, cutaneous lymphomas can be challenging to diagnose or even suspect because they mimic a variety of other inflammatory and neoplastic dermatological conditions. Support for non-invasive skin analysis methods like reflectance confocal microscopy and dermoscopy is still anecdotic. Practically speaking, a deeper and more comprehensive study with a larger number of cases focusing on the effective usefulness of non-invasive techniques should be taken into consideration because they have demonstrated the ability to identify macro and micro features supporting the clinical suspicion of lymphomas, as well as being useful for differential diagnosis and supporting the selection of the biopsy site. The author provides a brief and narrative synopsis of the reflectance confocal microscopy and dermoscopy characteristics of cutaneous lymphomas in this manuscript.

Comprehensive assessment of corneal microstructural changes following V4c implantable collamer lens surgery using in vivo confocal microscopy.

BACKGROUND: Implantable Collamer Lense (ICL) presents a viable alternative to conventional refractive surgeries, but their impact on corneal microstructure remains unclear. By employing in vivo confocal microscopy (IVCM), we examined changes in stromal and endothelial cells following the insertion of V4c ICLs, with the goal of enhancing post-surgical care and outcomes. METHODS: In this longitudinal investigation, we conducted detailed preoperative assessments on 103 eyes from 53 participants. Follow-up evaluations were carried out after surgery at set intervals: one day, one week, one month, three months, six months, and twelve months. We used IVCM to analyze changes in stromal and endothelial cells. To assess differences between pre- and post-surgery variables and to investigate correlations with age, axial length (AL), and spherical equivalent refraction (SER), we applied a repeated measures mixed-effects model, with statistical significance set at P < 0.05. RESULTS: No vision-threatening complications were reported post-surgery. Significant reductions in stromal cell density (SCD) were observed postoperatively, with anterior and mid- SCD reaching their lowest values at 3 months and posterior SCD at 1 month, remaining below baseline at 12 months. endothelial cell density (ECD) and percentage of hexagonal cells (PHC) decreased initially, recovering by 12 months. Conversely, endothelial cellular area (ECA) and coefficient of variation of cell size (CoV) increased postoperatively, with the most significant change at 1 week. Endothelial deposits were detected in 49 of 101 eyes on postoperative day 1, half of them were absorbed within 3 months post-surgery. Changes in posterior SCD were negatively related to AL, while AL, SER, lens thickness showed associated with endothelium changes. CONCLUSION: Our findings elucidate the corneal microstructural changes following V4c ICL implantation, particularly the significant early reductions in stromal and endothelial cell densities. We recommend careful management of viscoelastics during surgery to minimize endothelial deposits that may harm the endothelium. Enhanced early postoperative monitoring and these surgical adjustments can lead to improved surgical and post-surgical care, ultimately supporting better patient recovery.

Reflectance Confocal Microscopy and Dermoscopy for the Diagnosis of Solitary Hypopigmented Pink Lesions: A Narrative Review.

Diagnosing solitary pink skin lesions poses a significant challenge due to the scarcity of specific clinical and dermoscopic criteria. Several benign lesions, such as cherry angioma, clear cell acanthoma, dermal nevus, keloid, hypertrophic scar, and Spitz nevus, often exhibit similar clinical and dermoscopic features. This similarity extends to some malignant lesions, including basal cell carcinoma, actinic keratosis, and amelanotic melanoma, making differentiation difficult. Recent studies highlight the enhanced diagnostic accuracy of reflectance confocal microscopy (RCM), which offers increased sensitivity and specificity compared to dermoscopy alone for diagnosing skin cancer. This study aims to summarize the application of dermoscopy and RCM in distinguishing between benign and malignant pinkish-reddish skin lesions. The integration of RCM with traditional dermoscopic techniques improves the ability to accurately identify and differentiate these lesions. However, it is crucial to note that for any suspicious lesions, a final diagnosis must be confirmed through surgical excision and histopathological evaluation. This comprehensive approach ensures accurate diagnosis and appropriate treatment, highlighting the importance of combining advanced imaging techniques in clinical practice.

Idiopathic Distal Sensory Polyneuropathy and Fibromyalgia Syndrome: A Comparative Phenotyping Study.

INTRODUCTION: Painful idiopathic distal sensory polyneuropathy (IDSP) and fibromyalgia syndrome (FMS) are cryptogenic chronic pain syndromes. The contribution of small fibre pathology (SFP) in FMS remains controversial. This study aims to quantify small nerve pathology in participants with IDSP and FMS and identify relationships of SFP with sensory phenotypes. METHODS: In this study, 73 individuals (FMS: 25, IDSP: 23, healthy volunteers: 25) underwent comprehensive assessment, including neurological exams, questionnaires, sensory tests, and corneal confocal microscopy. RESULTS: IDSP participants displayed lower wind-up ratio (WUR) relative to FMS (p < 0.001), loss of function to thermal and mechanical stimuli and elevated neuropathy disability scores compared to FMS and healthy volunteers (all p < 0.001). FMS participants demonstrated gain of function to heat and blunt pressure pain responses relative to IDSP, and healthy volunteers (heat: p = 0.002 and p = 0.003; pressure: both p < 0.001) and WUR (both p < 0.001). FMS participants exhibited reduced corneal nerve fibre density (p = 0.02), while IDSP participants had lower global corneal nerve measures (density, branch density, and length) relative to healthy volunteers (all p < 0.001). Utilising corneal nerve fibre length, SFP was demonstrated in 66.6% of participants (FMS: 13/25; IDSP: 22/23). CONCLUSION: Participants with SFP, in both FMS and IDSP, reported symptoms indicative of small nerve fibre disease. Although distinctions in pain distributions are evident between individuals with FMS and IDSP, over 50% of participants between the two conditions displayed both a loss and gain of thermal and mechanical function suggestive of shared mechanisms. However, sensory phenotypes were associated with the presence of SFP in IDSP but not in FMS.

In people with painful idiopathic neuropathy (pain related to nerve damage where the cause of nerve damage is unknown), fibromyalgia syndrome (a long-term condition causing widespread pain), and healthy volunteers, the small nerve fibres of the peripheral nervous system, which may be involved in generating pain were assessed. These nerve fibres can be measured at the front of the eye (cornea) which can provide details on whether they are damaged in the body. The response to temperature, light touch, pressure and pinprick stimuli can also be used to determine if there is a loss or gain of sensation, which may contribute to pain. The aim of this study was to identify the degree of damage to these nerve fibres and to determine whether this damage is associated with a loss (cannot feel or requires more intense stimulus to feel) or gain (stimulus is felt earlier or is painful earlier at lower intensity) of sensory function. The pattern of loss or gain in sensory function is known as a sensory phenotype. It was found that people with painful idiopathic neuropathy had more severe nerve damage, loss of function to temperature and touch, and fewer small nerve fibres in the cornea compared to those with fibromyalgia syndrome and healthy volunteers. People with fibromyalgia syndrome were more sensitive to heat and pressure and had fewer corneal nerve fibres relative to healthy volunteers. The presence of corneal nerve fibre damage was associated with sensory phenotypes (types of sensation felt) in painful idiopathic neuropathy but not in fibromyalgia syndrome.

Use of in vivo confocal microscopy in suspected Acanthamoeba keratitis: a 12-year real-world data study at a Swedish regional referral center.

PURPOSE: To report real-world data (RWD) on the use of in vivo confocal microscopy (IVCM) in handling cases of suspected Acanthamoeba keratitis (AK) cases at a regional referral center during a 12-year period. METHODS: Retrospective study of patients with suspected AK presenting at a regional referral center for IVCM in Sweden from 2010 to 2022. Demographics, symptoms, outcomes, and clinical management were analyzed, and IVCM images were interpreted. RESULTS: Of 74 included patients with suspected AK, 18 (24%) were IVCM-positive, 33 (44%) were IVCM-negative, 15 had inconclusive IVCM results (20.2%), and 8 (11%) were referred for a second opinion based on IVCM, 4 of which were IVCM-positive (5.5%), yielding an overall IVCM-positive rate of 29.5%. Cultures were taken in 38 cases (51%) with only 2 cases (2.7%) culture-positive for AK. Of IVCM-negative cases, cultures were taken in 22 (67%) of cases and 100% of these were AK-negative. IVCM-positive cases had more clinic visits (median 30, P = 0.018) and longer follow-up time (median 890 days, P = 0.009) than IVCM-negative patients, while visual acuity improvement did not differ (P > 0.05). Of IVCM-positive cases, 10 (56%) underwent surgery despite prior anti-amoebic treatment, and 14 (78%) had 3 or more IVCM examinations during follow-up, with cysts (100%), dendritic cells (89%) and inflammatory infiltrate (67%) as the most prevalent features. Longitudinal IVCM indicated improvement in cysts, dendritic cells and subbasal nerves with treatment, while clinical resolution was not always consistent with complete absence of cysts. CONCLUSIONS: In a real-world setting, IVCM has a high reliability in classifying AK-negative cases, while IVCM detects AK-positive cases more frequently than the gold-standard culture method, leading to its preferential use over the culture method where time or resources are limited. Despite this, a subset of cases are IVCM-inconclusive, the clinical course of referred patients is long requiring many hospital visits, and visual acuity in most cases does not improve with medical treatment alone. Information sharing across centers and standardization of referral and diagnostic routines is needed to exploit the full potential of IVCM in AK patient management.

In Vivo Confocal Microscopy in Avellino Corneal Dystrophy.

Corneal dystrophies are a group of rare genetic eye disorders characterized by the accumulation of abnormal material in different layers of the cornea, potentially leading to vision impairment. In vivo confocal microscopy (IVCM) is an emerging non-invasive imaging and diagnostic tool that helps study the ocular surface microstructure. This case report examines the clinical characteristics of Avellino corneal dystrophy in a young patient through the use of slit lamp examination, IVCM, and optical coherence tomography (OCT) in order to assess the effectiveness of these non-invasive tests as diagnostic tools.

An Imaging-Based Assay to Measure the Location of PD-1 at the Immune Synapse for Testing the Binding Efficacy of Anti-PD-1 and Anti-PD-L1 Antibodies.

PD-1 is an immune checkpoint on T cells. Antibodies to PD-1 or its ligand PD-L1 are gaining popularity as a leading immunotherapy approach. In the US, 40% of all cancer patients will be treated with anti-PD-1 or anti-PD-L1 antibodies but, unfortunately, only 30% will respond, and many will develop immune-related adverse events. There are nine FDA-approved anti-PD-1/PD-L1 antibodies, and approximately 100 are in different stages of clinical development. It is a clinical challenge to choose the correct antibody for a given patient, and this is critical in advanced malignancies, which often do not permit a second-line intervention. To resolve that, an in vitro assay to compare the performance of the different anti-PD-1/PD-L1 antibodies is not only a critical tool for research purposes but also a possible tool for personalized medicine. There are some assays describing the binding affinity and function of anti-PD-1/PD-L1 antibodies. However, a significant limitation of existing assays is that they need to consider the location of PD-1 in the immune synapse, the interface between the T cell and tumor cells, and, therefore, ignore a critical component in its biology. To address this, we developed and validated an imaging-based assay to quantify and compare the ability of different anti-PD-1/PD-L1 antibodies to remove PD-1 from the immune synapse. We correlated that with the same antibodies' ability to increase cytokine secretion from the targeted cells. The strong correlation between PD-1 location and its function in vitro and in vivo within the antibody treatment setting validates this assay's usability, which is easily recordable and straightforward. Key features • Live-cell imaging quantifies and compares how anti-PD-1 and anti-PD-L1 antibodies disrupt PD-1 localization, causing the removal of PD-1 during immune synapse formation. • Hao et al. [1] validated the protocol, and the findings were extended to a live confocal microscopy method. • It requires a Zeiss LSM 900 confocal microscope and appropriate imaging software and is optimized for the latest version of Zen Blue. • Anti-PD-1 antibodies are commonly used in cancer therapies, and this protocol optimizes the analysis of their effectiveness.

A Method for Quantitative Spatial Analysis of Immunolabeled Fibers at Regenerative Electrode Interfaces.

BACKGROUND: Regenerative electrodes are being explored as robust peripheral nerve interfaces for neuro-prosthetic control and sensory feedback. Current designs differ in electrode number, spatial arrangement, and porosity which impacts the regeneration, activation, and spatial distribution of fibers at the device interface. Knowledge of sensory and motor fiber distributions are important in optimizing selective fiber activation and recording. NEW METHOD: We use immunofluorescence and confocal microscopy to visualize and segment whole nerve for spatial analysis of immunolabeled fibers at regenerative electrode interfaces. RESULTS: This method was implemented to characterize motor fiber distribution within 3 macro-sieve electrode regenerated (MSE), 3 silicone-conduit regenerated, and 3 unmanipulated control rodent sciatic nerves. Total motor fiber counts were 1485 [SD: +/- 50.11], 1899 [SD: +/- 359], and 5732 [SD: +/- 1410] for control, MSE, and conduit nerves respectively. MSE motor fiber distributions exhibited evidence of deviation from complete spatial randomness and evidence of dispersion and clustering tendencies at varying scales. Notably, MSE motor fibers exhibited clustering within the central portion of the cross section, whereas conduit regenerated motor fibers exhibited clustering along the periphery. COMPARISON WITH EXISTING METHODS: Prior exploration of fiber distributions at regenerative interfaces was limited to either quadrant-based density analysis of randomly sampled subregions or qualitative description. This method extends existing sample preparation and microscopy techniques to quantitatively assess immunolabeled fiber distributions within whole nerve cross-sections. CONCLUSIONS: This method is an effective way to examine the spatial arrangement of immunolabeled fiber subsets at regenerative electrode interfaces, enabling robust assessment of fiber distributions relative to electrode arrangement.

Artificial targeting of the NEIL1 DNA glycosylase to the mitochondria.

The human NEIL1 DNA glycosylase is one of 11 mammalian glycosylases that initiate base excision repair. While substrate preference, catalytic mechanism, and structural information of NEIL1's ordered residues are available, limited information on its subcellular localization, compounded by relatively low endogenous expression levels, have impeded our understanding of NEIL1. Here, we employed a previously developed computational framework to optimize the mitochondrial localization signal of NEIL1, enabling the visualization of its specific targeting to the mitochondrion via confocal microscopy. While we observed clear mitochondrial localization and increased glycosylase/lyase activity in mitochondrial extracts from low-moderate NEIL1 expression, high NEIL1 mitochondrial expression levels proved harmful, potentially leading to cell death.

Multimodal imaging and histopathological evaluation in silicone oil keratopathy.

PURPOSE: To describe features in silicone oil keratopathy using multimodal imaging and histopathological examination. METHODS: Case report. RESULT: A 21-year-old male developed right corneal decompensation in the heavy SO (HSO)-filled eye. The patient underwent an initial lensectomy, pars plana vitrectomy (PPV) and HSO tamponade due open-globe injury with corneal wound, lens damage and in two retained intravitreal glass foreign bodies, followed by a revisional PPV with HSO tamponade due to tractional detachment associated with proliferative vitreoretinopathy and epiretinal membrane. One month after the removal of HSO, ophthalmic examination of the right eye showed corneal decompensation. The AS-OCT showed corneal thickening, intrastromal scattered hyperreflective dots and large rounded/oval hyporeflective space; the latter were suggestive of emulsified HSO microbubbles and larger bubbles, respectively. In vivo confocal microscopy showed multiple presumed SO-related corneal changes, including hyper-reflective fibrotic changes in the basal epithelium, reduced density ans altered morphology of keratocytes cell population, increased pleomorphism and polymegathism of the endothelium with reduced endothelial cell, and presence of inflammatory cells. The patient underwent a penetrating keratoplasty, pupilloplasty and retropupillary iris-claw IOL implantation. The histopathological examination of the host corneal button showed Descemet's membrane irregularity and thickened corneal stroma with focal intrastromal silicone oil vacuoles, surrounded by macrophages. CONCLUSION: We described for the first time intrastromal hyperreflective dots as a sign associated with SO-related keratopathy. Moreover, this case report supports the ability of emulsified SO to penetrate the cornea inducing a local low-grade chronic inflammation.

ColocZStats: a z-stack signal colocalization extension tool for 3D slicer.

Confocal microscopy has evolved to be a widely adopted imaging technique in molecular biology and is frequently utilized to achieve accurate subcellular localization of proteins. Applying colocalization analysis on image z-stacks obtained from confocal fluorescence microscopes is a dependable method of revealing the relationship between different molecules. In addition, despite the established advantages and growing adoption of 3D visualization software in various microscopy research domains, there have been few systems that can support colocalization analysis within a user-specified region of interest (ROI). In this context, several broadly employed biological image visualization platforms are meticulously explored in this study to understand the current landscape. It has been observed that while these applications can generate three-dimensional (3D) reconstructions for z-stacks, and in some cases transfer them into an immersive virtual reality (VR) scene, there is still little support for performing quantitative colocalization analysis on such images based on a user-defined ROI and thresholding levels. To address these issues, an extension called ColocZStats (pronounced Coloc-Zee-Stats) has been developed for 3D Slicer, a widely used free and open-source software package for image analysis and scientific visualization. With a custom-designed user-friendly interface, ColocZStats allows investigators to conduct intensity thresholding and ROI selection on imported 3D image stacks. It can deliver several essential colocalization metrics for structures of interest and produce reports in the form of diagrams and spreadsheets.

Corneal nerve changes by anti-glaucoma medications examined by in vivo confocal microscopy.

AIM: To evaluate the effects of antiglaucoma eye drops on corneal nerves by in vivo confocal microscopy (IVCM). METHODS: This study comprised 79 patients diagnosed with glaucoma and 16 healthy control individuals. Among the glaucoma patients, 54 were treated with medication, while 25 remained untreated. Central corneal images were evaluated by IVCM, and then ACCMetrics was used to calculate the following parameters: corneal nerve fiber density (CNFD), branch density (CNBD), fiber length (CNFL), total branch density (CTBD), fiber area (CNFA), fiber width (CNFW), and fractal dimension (CNFrD). The correlation between IVCM parameters and drugs was evaluated using non-parametric measurements of Spearman's rank correlation coefficient. RESULTS: The CNFD was reduced in glaucoma groups compared to healthy subjects (P<0.01). Patients using anti-glaucoma medications exhibited poorer confocal parameters compared to untreated patients. As the number of medications and usage count increased, CNFD, CNBD, CNFL, CTBD, CNFA, and CNFrD experienced a decline, while CNFW increased (all P<0.01). For the brinzolamide-therapy group, there was a significant decrease in CNFD and CNFL compared to the other monotherapy groups (P<0.001). In the absence of medication, CNFD in males was lower than that in females (P<0.05). Among patients under medication therapy, CNFD remained consistent between males and females. CONCLUSION: Antiglaucoma eye drops affect the microstructure of corneal nerves. IVCM and ACCMetrics are useful tools that could be used to evaluate the corneal nerve changes.

Photographic scale for the characterization of actinic keratosis through reflectance confocal microscopy: a quantitative approach to cellular transformation.

Background: Actinic keratosis (AK) is a highly prevalent pre-cancerous skin lesion that often leads to cutaneous squamous cell carcinoma. There are different stages of evolution of the disease and several features that characterize keratosis. This study aimed to develop a qualitative and quantitative visual diagnostic tool to facilitate the identification of the characteristics and severity of the main cellular attributes of AK and to show its applicability in evaluating the evolution or treatment through image analysis. Methods: Literature research on the main scientific databases and in the institute's database was carried out to gather all the different levels of cellular transformation. To validate the scale, a preliminary characterization study was carried out with 21 subjects who had clinically diagnosed AK lesions to classify the attributes in each skin layer and test the accuracy of the diagnosis of the scale. Afterward, and to show the possibility of a follow-up with a topical treatment, the subjects were divided into two treatment groups, receiving either a cream formulation containing retinoic acid, or a placebo formula. The evaluation was carried out through confocal reflectance microscopy and a digital camera with dermoscopic quality before and after 90 days of treatment. Results: A table detailing the 18 attributes of AK, and a photographic scale containing RCM images graded by scores established for each characteristic and the frequency of spreading were developed. The results of the validation presented good repeatability, correlation with clinical evaluation, and capacity for differentiating treatments demonstrated by the significant improvement after topical treatment by the reduction of the score for 10 out of the 18 attributes. The preliminary study, evaluated by the detailed transformation scale highlights important differences in the subclinical approach that allows a deeper evaluation of the aspects of the lesion's re-incidence even after fully treated skin sites. Conclusion: This study brings an innovative method based on RCM, to assist in the quantification of cell transformation level, provide early diagnosis, and deliver a powerful treatment evaluation tool to provide smoother treatment, as well as prevent re-incidence in the cases.

Noninvasive Technologies for the Diagnosis of Squamous Cell Carcinoma: A Systematic Review and Meta-Analysis.

Early cutaneous squamous cell carcinoma (cSCC) diagnosis is essential to initiate adequate targeted treatment. Noninvasive diagnostic technologies could overcome the need of multiple biopsies and reduce tumor recurrence. To assess performance of noninvasive technologies for cSCC diagnostics, 947 relevant records were identified through a systematic literature search. Among the 15 selected studies within this systematic review, 7 were included in the meta-analysis, comprising of 1144 patients, 224 cSCC lesions, and 1729 clinical diagnoses. Overall, the sensitivity values are 92% (95% confidence interval [CI] = 86.6-96.4%) for high-frequency ultrasound, 75% (95% CI = 65.7-86.2%) for optical coherence tomography, and 63% (95% CI = 51.3-69.1%) for reflectance confocal microscopy. The overall specificity values are 88% (95% CI = 82.7-92.5%), 95% (95% CI = 92.7-97.3%), and 96% (95% CI = 94.8-97.4%), respectively. Physician's expertise is key for high diagnostic performance of investigated devices. This can be justified by the provision of additional tissue information, which requires physician interpretation, despite insufficient standardized diagnostic criteria. Furthermore, few deep learning studies were identified. Thus, integration of deep learning into the investigated devices is a potential investigating field in cSCC diagnosis.

Usability of a smartphone-compatible, confocal micro-endoscope for cervical cancer screening in resource-limited settings.

BACKGROUND: More efficient methods to detect and treat precancerous lesions of the cervix at a single visit, such as low-cost confocal microscopy, could improve early diagnosis and hence outcomes. We piloted a prototype smartphone-compatible confocal micro-endoscope (SCME) among women presenting to a public cervical cancer screening clinic in Kampala, Uganda. We describe the piloting of the SCME device at an urban clinic used by lower cadre staff. METHODS: We screened women aged 18 and 60 years, who presented for cervical cancer screening at the Kawempe National Referral Hospital Kampala, and evaluated the experience of their providers (nurses). Nurses received a 2-day training by the study doctors on how to use the SCME, which was added to the standard Visual Inspection with Acetic acid (VIA)-based cervical cancer screening. The SCME was used to take colposcopy images before and after VIA at positions 12 and 6 O'clock if VIA negative, and on precancer-suspicious lesions if VIA positive. We used questionnaires to assess the women's experiences after screening, and the experience of the nurses who operated the SCME. RESULTS: Between November 2021 and July 2022, we screened 291 women with a median age of 36 years and 65.7% were HIV positive. Of the women screened, 146 were eligible for VIA, 123 were screened with the SCME, and we obtained confocal images from 103 women. Of those screened with the SCME, 60% found it comfortable and 81% were willing to screen again with it. Confocal images from 79% of the women showed distinguishable cellular features, while images from the remaining 21% were challenging to analyze. Nurses reported a mean score of 85% regarding the SCME's usefulness to their work, 71% regarding their satisfaction and willingness to use it again, 63% in terms of ease of use, and 57% concerning the ease of learning how to operate the SCME. CONCLUSION: Our findings demonstrate the feasibility of using the SCME by lower cadre staff in low-resource settings to aid diagnosis of precancerous lesions. However, more work is needed to make it easier for providers to learn how to operate the SCME and capture high-quality confocal images.

Melatonin production improves Senegalese sole sperm motility at night, but fails as a supplement during cryopreservation.

Melatonin is a powerful antioxidant present in fish seminal plasma. This study aimed to understand melatonin's endogenous and exogenous effects on first-generation Senegalese sole sperm quality for sperm management applications. In the first experiment, samples were collected at mid-light (ML) and mid-dark (MD) daytimes, to evaluate the effects on sperm motility. In a second experiment, using confocal microscopy and melatonin-FITC, spermatozoa permeability to melatonin was evaluated and, after showing that it enters the nucleus and mitochondria by passive diffusion, exogenous melatonin toxicity and antioxidant potential during a cryopreservation assay were performed. The toxicity assay tested different melatonin concentrations (0.01, 0.1, 1, and 10 mM) and exposure times (3, 5, 15 and 30 min), and sperm motility parameters were measured (TM, PM, VCL, VSL, LIN) using CASA system. The best conditions (0.1 and 10 mM) were selected for the cryopreservation assay, and a set of post-thaw sperm quality analyses were performed (motility, viability, reactive oxygen species, lipid peroxidation, and DNA fragmentation). The motility analyzed at ML and MD showed significant differences in all parameters, mainly on velocities (VCL, VSL, VAP), that were significantly higher at MD. Supplemented melatonin did not influence spermatozoa motility, MDA content or DNA fragmentation, although a lower percentage of viable cells was obtained on the 10 mM treatment. Altogether, Senegalese sole spermatozoa motility was enhanced at night, putatively by endogenous melatonin through direct or indirect mechanisms, whereas supplemented melatonin did not confer extra protection during cryopreservation.

Application and progress of corneal confocal microscopy in the evaluation of diabetes-related peripheral neuropathy.

Diabetic peripheral neuropathy (DPN) is one of the most common chronic complications of diabetes. As a new detection method for DPN, corneal confocal microscopy (CCM) is characterised by rapid, non-invasive, sensitive, and quantitative characteristics, as well as good repeatability. By detecting changes in the corneal nerves, DPN can be diagnosed early, and the severity of neuropathy evaluated. It is currently an ideal DPN evaluation method and has good clinical application prospects. This paper reviews the application and progress of CCM in the evaluation of DPN and summarises the evaluation methods of CCM, corneal nerve, and DPN to provide new ideas for the clinical diagnosis and treatment of DPN.

[Comparative assessment of the state of the cornea after YAG-laser interventions on anterior segment structures]. / Sravnitel'naya otsenka sostoyaniya rogovitsy posle YAG-lazernykh vmeshatel'stv na strukturakh perednego segmenta glaza.

YAG-laser interventions are associated with the risk of complications, including in the cornea. PURPOSE: This study evaluates the condition of the cornea after laser discission (LD) of secondary cataracts (SC) and laser iridectomy (LI) using corneal confocal microscopy (CCM). MATERIAL AND METHODS: Group 1 included patients with the diagnosis "Pseudophakia, secondary cataract", they underwent LD of SC. Patients of group 1 were divided into 2 subgroups depending on the initial state of the cornea: group 1A included patients with unaltered corneas; group 1B - with changes in the corneas. Group 2 included patients diagnosed with angle-closure glaucoma (ACG) or suspected ACG, they underwent LI. CCM was performed on the Heidelberg HRT-III system. Laser treatment was performed using the Nd:YAG-laser LPULSA SYL-9000, λ=1.064 µm. RESULTS: Immediately after treatment, subgroup 1A exhibited singular hyperreflective deposits and negligible endothelial cell loss (ECL). After 1 month, CMM findings revealed no changes in this subgroup. In subgroup 1B, a post-LD reduction in endothelial cell density led to increased polymegathism, decreased pleomorphism, heightened endothelial cell nucleus reflectivity, and moderate hyperreflective deposits after 1 month. In the second group, significant hyperreflective deposits of various sizes, increased nucleus reflectivity, and notable endothelial cell density reduction were observed immediately and 1 month after LI. CONCLUSION: The results of this study show that the possibility of developing corneal complications after photo destructive laser interventions is to a certain extent related to the initial state of the cornea. The risk of developing corneal damage increases with decreasing distance between the cornea and the irradiated structure. An increase in the level of laser radiation energy and its total values also contributes to damage to the cornea, which is possible with dense secondary cataracts and thick irises.

[Purulent scleritis following penetrating keratoplasty for acanthamoeba keratitis]. / Gnoinyi sklerit posle skvoznoi keratoplastiki po povodu akantamebnogo keratita.

The problem of treating purulent scleral infections, rare but extremely severe complication of ophthalmic surgeries, remains unresolved. This article presents a case of successful surgical treatment of purulent scleritis - interlamellar scleral abscess - that developed in a patient after repeat penetrating keratoplasty performed due to infectious lysis of the transplant. Although the first keratoplasty was performed for acanthamoeba keratitis, there were no signs of acanthamoeba invasion in the transplant at the time of the second surgery. Scleritis manifested as an infiltrate with pus penetrating the anterior chamber and development of keratoiridocyclitis. During surgery, the abscess cavity was opened, irrigated with an antiseptic solution, and drained into the subconjunctival space; the anterior chamber was irrigated with balanced salt solution through a separate paracentesis. No infection recurrences were noted in the postoperative period and the corneal transplant remained clear.