Cell Density In Patients Research Articles

Deep-learning based analysis of in-vivo confocal microscopy images of the subbasal corneal nerve plexus’ inferior whorl in patients with neuropathic corneal pain and dry eye disease

PurposeTo evaluate and compare subbasal corneal nerve parameters of the inferior whorl in patients with dry eye disease (DED), neuropathic corneal pain (NCP), and controls using a novel deep-learning-based algorithm to analyze in-vivo confocal microscopy (IVCM) images. MethodsSubbasal nerve plexus (SNP) images of the inferior whorl of patients with DED (n = 49, 77 eyes), NCP (n = 14, 24 eyes), and controls (n = 41, 59 eyes) were taken with IVCM and further analyzed using an open-source artificial intelligence (AI)-based algorithm previously developed by our group. This algorithm automatically segments nerves, immune cells, and neuromas in the SNP. The following parameters were compared between groups: nerve area density, average nerve thickness, average nerve segment tortuosity, junction point density, neuroma density, and immune cell density. Results160 eyes of 104 patients (63 % females), aged 56.8 ± 15.4 years, were included. The mean nerve area density was significantly lower in the DED (P = 0.012) and NCP (P < 0.001) groups compared to the control group. The junction point density was lower in the NCP group compared with control (P = 0.001) and DED (P = 0.004) groups. The immune cell density was higher in the DED group compared with controls (P < 0.001). ConclusionsDeep-learning-based analysis of IVCM images of the corneal SNP inferior whorl distinguished a decreased mean nerve area density in patients with DED and NCP compared with controls and an increased immune cell density in patients with oGVHD- and SS-associated DED. These findings suggest that the inferior whorl could be used as landmark to distinguish between patients with DED and NCP.

The effect of irrigation and aspiration on the corneal endothelial cell density in patients undergoing Implantable Collamer Lens with a central hole implantation for myopia correction.

In the conventional technique, viscoelastic agents are typically rinsed away with balanced salt solution (BSS), but it may lead to a series of complications such as viscoelastic residue, anterior chamber instability and intraoperative TICL rotation. The utilization of irrigation and aspiration (I/A) has been shown to be effective in maintaining anterior chamber stability, reducing the incidence of postoperative high intraocular pressure, and minimizing postoperative fundus complications. However, there is a lack of previous studies investigating the impact of I/A on corneal endothelial cells during ICL implantation. The objective of this study was to examine the effect of I/A on corneal endothelial cells in patients undergoing myopia correction through implantation of Implantable Collamer Lens with a central hole (V4c ICL). A retrospective selection was made of 344 eyes from 172 patients who underwent V4c ICL implantation and I/A to remove viscoelastic agent from the anterior chamber between 2021 and 2022. The intraocular pressure (IOP) was measured at 1h, 2h and 3h after surgery. Corneal endothelial cell density (ECD), coefficient of variation in cell size (CV), standard deviation of cell area (SD), and percentage of hexagonal cells (HEX) were evaluated at 1week postoperatively to assess corneal endothelial cells. The first two represent polymegethism or morphological variation, while the third parameter represents the degree of polymorphism of the corneal endothelial cells. Electronic medical records were utilized for data collection purpose. All surgeries proceeded without complications. The IOP was 16.50 ± 3.42mmHg (range: 11.5-22.3mmHg) prior to surgery and increased to 21.25 ± 5.61mmHg (range: 9.5-34.8mmHg), 19.85 ± 5.18mmHg (range: 11.4-36.2mmHg) and finally settled at an average of 18.81 ± 4.57mmHg (range: 10.1-38.8mmHg) at the respective time points of 1h, 2h and 3h after surgery. The preoperative ECD was recorded as being approximately 3004 ± 295 cell/mm2, which exhibited a marginal decreased of 1.17% postoperatively, resulting in an average ECD value of 2969 ± 303 cell/mm2 one week after surgery (P = 0.12). Similarly, the preoperative CV was determined as 31.10 ± 3.78%, and it experienced a slight reduction with an average CV value of 30.74 ± 3.77% at week after surgery (P = 0.21). And, the preoperative SD was reported as 104.76 ± 17.26, and it remained virtually unchanged with an average SD value of 104.85 ± 18.75 at one week after surgery (P = 0.95). The preoperative HEX was calculated as 55.38 ± 8.94%, and it remained its stability with an average HEX value of 55.45 ± 8.73% one week after surgery (P = 0.92). The utilization of I/A led to a slight decrease in postoperative ECD when compared to conventional surgical techniques. Nevertheless, the reduction in ECD remained within acceptable limits, taking into accout the avervantaged it offered, such as stabilization of the anterior chamber and decreased occurrence of viscoelastic residue after surgery. It is challenging to anticipate the long-term safety of corneal endothelial cells based on current short-term studies. However, this study provides a valuable reference indicating that neither anterior chamber irrigation nor I/A aspiration have an adverse impact on the safety of corneal endothelial cells in the short term. Further research is imperative to enhance our understanding of their effects over an extended period.

Corneal complications of ocular GvHD

Corneal fluorescein staining is one of the tests recommended to diagnose and grade ocular graft versus host disease (GvHD) according to the International Chronic Ocular GvHD Consensus Group. Despite superficial punctate keratopathy is the most common corneal manifestation, patients with more severe stages may develop corneal neovascularization, sterile corneal ulceration and even perforation. Previous in vivo confocal microscopy (IVCM) studies reported significant microstructural changes in the cornea of patients with ocular GvHD, including a higher density of dendritic cells and globular immune cells, a network of hyper‐reflective activated keratocytes, and a lower density and higher tortuosity of sub‐basal corneal nerves. Dendritic cells act as antigen‐presenting cells, and play a key role in ocular surface immune homeostasis. Thus, their increased density may be indicative of immune activation and inflammation of the ocular surface in patients with ocular GvHD. Interestingly, Kheirkhah et al. reported higher dendritic cell density in patients with DED owing to systemic immune disease (i.e., Sjögren syndrome and ocular GvHD) compared to patients with DED of other origins. Recently, our group employed a fully automated IVCM analysis system to compare corneal sub‐basal nerve plexus in patients with DED owing to both ocular GvHD and Sjögren syndrome and in healthy control subjects. Although overall patients with DED showed lower density of nerve fibres and branches, shorter nerve fibres and higher fibre width compared to healthy controls, no significant difference was observed between patients with ocular GvHD and Sjögren syndrome for all IVCM metrics. In agreement with this observation, other studies reported no significant difference of IVCM parameters in DED patients with and without ocular GvHD after adjusting for clinical severity of dry eye. This suggests that the ocular surface changes observed by IVCM in ocular GvHD may be possibly reflective of the local disease severity rather than the underlying systemic process.

Comparison of central corneal thickness and endothelial cell count among diabetic retinopathy and non-diabetic cases

: Diabetes is associated with alterations in the structure and functions of corneal endothelial cells. Current research has been carried out to compare the central corneal thickness and endothelial cell density in patients with diabetic retinopathy (DR) and non-diabetic (ND) individuals. : A Cross Sectional comparative research was carried out in a tertiary eye care hospital, over the period of two years from April 2021 to March 2023. Among 160 study participants, 80 cases with diabetic retinopathy (DR) and 80 non-diabetic (ND) cases as age matched control have been selected. Thorough Ophthalmic assessment was done. We have included only retinopathy cases as they have longer duration of diabetis and poorer metabolic control, for better comparison of the endothelium parameters with non-diabetics. Specular microscopy has been done in all cases for endothelial cell count evaluation and thickness of cornea has been estimated by Pachymeter. Statistical analysis was carried out by students‘t’ test by comparing the variables between two groups. : P-value was not significant for the mean age and sex distribution in the two groups. Mean endothelial cell density was lesser (2512.12±260.23cells/mm2) in DR than in ND group (2699.10±95.68cells/mm2). Mean central corneal thickness was greater (522.65±36.56μm) in DR than in ND group (486.50±18.67μm) (P&amp;#60;0.05). Also, the Co-efficient of variation percentage was more whereas the percentage of hexagonality was found to be statistically less in DR than ND group. Among DR patients, endothelial cell density was significantly reduced and central corneal thickness considerably raised when compared with ND. Our results also suggest that poor metabolic control and advanced Diabetic Retinopathy are risk factors for developing keratopathy.

Assessment of Corneal Endothelial Cell Density in Patients with Dry Eye Disease

Assessment of Corneal Endothelial Cell Density in Patients with Dry Eye Disease

Effect of intravitreal ranibizumab on fibrovascular membranes in patients with proliferative diabetic retinopathy.

To assess the effects of intravitreal ranibizumab (IVR) on angiogenesis and glial activity of the fibrovascular membrane (FVM) in patients with proliferative diabetic retinopathy (PDR). Forty-two eyes from 42 patients with PDR requiring vitrectomy were included and divided into two groups: control group (n=16) did not receive IVR, while IVR group (n=26) underwent IVR 5d before vitrectomy. FVM specimens were collected by the same surgeon during the interventions. Histopathological morphology was examined by hematoxylin-eosin (H-E) staining and cell densities in the FVM was assessed. Microvessels were outlined by immunohistochemical staining of CD31 and microvessel density (MVD) assessed as an index of FVM angiogenesis. Dual-color immunofluorescence staining, and confocal microscopy was used to detect co-localization and relative expression levels of glial fibrillary acidic protein (GFAP) and α-smooth muscle actin (α-SMA) as markers of glial-mesenchymal transition (GMT). The GMT index (GI; ratio of relative GFAP/α-SMA expression) was used to semi-quantify the degree of GMT or glial activity of FVMs. H-E staining showed similar vascularization in both groups, with microvessels and scattered stromal cells in the matrix. Infiltrated cell densities did not differ significantly between the two groups (P>0.05). The MVD of the IVR group (130.62±15.46/mm2) was significantly lower than that of the controls (142.25±19.16/mm2, P<0.05). In both groups, all sections were strongly immunostained for GFAP and α-SMA. The Pearson's correlation coefficients (PCC) of intensity of automated pixel count of two markers indicated GFAP and α-SMA co-stained well and GMT participated in the remolding of FVMs in PDR. The mean relative GFAP expression in the IVR group was significantly lower, whereas that of α-SMA was significantly higher than in controls (P<0.05). GI in the IVR group (1.10±0.10) was significantly lower than in the controls (1.21±0.12, P<0.05). IVR can reduce angiogenesis, glial activity of FVM and promote glial-fibrotic transformation by reducing MVD and promoting GMT but does not decrease the cell density in patients with PDR.

Assessment of Corneal Endothelial Cell Density in Patients with Dry Eye Disease

Assessment of Corneal Endothelial Cell Density in Patients with Dry Eye Disease

Subclinical Corneal Nerve Fiber Damage and Immune Cell Activation in Systemic Lupus Erythematosus: A Corneal Confocal Microscopy Study.

PurposeThe purpose of this study was to evaluate the utility of corneal confocal microscopy (CCM) in identifying small nerve fiber damage and immune cell activation in patients with systemic lupus erythematosus (SLE).MethodsThis cross-sectional comparative study included 39 consecutive patients with SLE and 30 healthy control participants. Central corneal sensitivity was assessed using a Cochet-Bonnet contact corneal esthesiometer and a laser scanning CCM (Heidelberg, Germany) was used to quantify corneal nerve fiber density (CNFD), nerve branch density (CNBD), nerve fiber length (CNFL), and Langerhans cell (LC) density.ResultsAge was comparable among patients with SLE (33.7 ± 12.7) and controls (35.0 ± 13.7 years, P = 0.670) and the median duration of disease was 3.0 years (2.0–10.0 years). CNBD (P = 0.003) and CNFL (P = 0.019) were lower and mature LC density (P = 0.002) was higher, but corneal sensitivity (P = 0.178) and CNFD (P = 0.198) were comparable in patients with SLE compared with controls. The SELENA-SLEDAI score correlated with CNFD (ρ = −0.319, P = 0.048) and CNFL (ρ = −0.373, P = 0.019), and the total and immature LC densities correlated with CNBD (ρ = −0.319. P = 0.048, and ρ = −0.328, P = 0.041, respectively). Immature LC density was higher (P = 0.025), but corneal sensitivity and nerve fiber parameters were comparable between patients with (33%) and without neuropsychiatric symptoms and SLE.ConclusionsCorneal confocal microscopy identifies distal corneal nerve fiber loss and increased immune cell density in patients with SLE and corneal nerve loss was associated with disease activity.Translational RelevanceCorneal confocal microscopy may enable the detection of subclinical corneal nerve loss and immune cell activation in SLE.

Corneal endothelial cell density in patients receiving chemotherapy

Purpose This study aimed to determine if the corneal endothelium was affected by chemotherapy. Methods Chemotherapy patients were recruited to undergo specular microscopy before treatment and again at 1- and 2-year follow-up visits. One eye per patient, per follow-up, was selected for comparison to baseline. Results Forty-six volunteers completed baseline and at least one follow-up assessment. From 51 eyes, there was no significant change in endothelial cell density for 41 eyes assessed at one year (MD = 0.73%, 95% CI −1.33 to 2.78%) and 18 eyes at two years (MD = 0.31%, 95% CI −3.53 to 4.15%). Conclusion Although other studies have shown that chemotherapy can adversely affect the corneal epithelium, this study showed no measurable change in endothelial cell density.

Is Pterygium Morphology Related to Loss of Corneal Endothelial Cells? A Cross-Sectional Study.

PurposeTo determine the difference in mean corneal endothelial cell density (CECD) between the healthy and diseased eyes of the patients with unilateral pterygium with different morphology patterns by using a non-contact specular microscope (SP2000: Topcon Corporation, Japan) and to find out any relationship between severity of pterygium and daily sunlight exposure with the CECD loss.MethodsA descriptive cross-sectional prospective study was carried out at Al-Shifa Trust Eye Hospital (ASTEH), Rawalpindi, Pakistan from 21st January 2019 to 22nd January 2020. Two hundred eyes (n= 100 patients) of age range18 -68 years with unilateral pterygium were selected. Necessary demographic data and essential variables like age, smoking status, occupation, and daily direct sunlight exposure were determined. The severity of pterygium (grading) based on its morphology was determined by slit-lamp examination. CECD of each patient was carried out using a non-contact Specular Microscope. The healthy eye (without pterygium) of a patient was considered as control.ResultsThe age range in this study was 18–68 years, with a mean age of 43.80 ± 24.37 years with a male to female ratio of 1.6:1 (62.00% males vs 38% females). Out of six occupations, the most common occupation was labour/construction work (n=31) followed by farming (n=27). The study reported a mean corneal endothelial cell density (CECD) of 2411.61±143.64 vs 2751.41 ± 123.674 cells/mm2 in diseased and normal eyes, respectively (p-value = 0.0001). CECD was lower in grade 3 pterygium compared to less severe pterygium { grade 3 (Fleshy) =2261 cells/mm2 vs grade 2 (Intermediate)= 2413 cells/mm2 vs grade 1 (Atrophic)=2459 cells/mm2} although this difference between the groups was not found to be significant (p=0.065). No significant association between sunlight exposure and CECD loss was observed (p=0.065).ConclusionThis study concluded that the mean corneal endothelial cell density in patients with unilateral pterygium using a non-contact specular microscope were 2411.61±143.64 cells/mm2.

Comparison of Central Corneal Thickness and Endothelial Cell Density in Patients with Various Types of Glaucoma and Patients without Glaucoma: A Case-control Study

Introduction: Corneal affection in glaucoma patients is very high due to various risk factors that may lead to unforeseen and unplanned deleterious effects on the cornea leading to vision loss. Accurate intraocular pressure determination requires corneal thickness measurement and uncontrolled intraocular pressures, use of long term medication, intra ocular surgeries including cataract and glaucoma shunt surgeries may cause significant endothelial loss. Therefore, adequate planning and management and follow-up in these patients is required weighing all the long term consequences and emphasis the need for implementing adequate precautions. Aim: To evaluate and compare the Central Corneal Thickness (CCT) and corneal Endothelial Cell Density (ECD) in patients with glaucoma and without glaucoma by specular microscopy. Materials and Methods: A case-control study conducted in a Shekar Eye Hospital, Bengaluru, Karnataka, India, was conducted from August 2016 till April 2018 on a total 182 eyes from 182 patients were evaluated comprising of 91 cases and 91 controls. All participants underwent a detailed ophthalmological examination including slit lamp biomicroscopy, Intraocular Pressure (IOP) measurement, and CCT and ECD measurement by Tomey EM 3000 noncontact specular microscopy. The Statistical analysis was completes using Statistical Package For the Social Sciences (SPSS) 18.0 and R environment version 3.2.2 software’s. Results: The CCT of the glaucoma patients and the controls showed no significant difference (p-value=0.172). The CCT was comparatively thicker in Ocular Hypertension (OHTN) patients and thinner in Normal Tension Glaucoma (NTG) however statistical significance was not established. The mean ECD of controls was 2509.05±298.48 (cells/mm3) and that of cases was 2465.68±392.91 (cells/mm3) (p-value=0.404). The difference in the ECD amongst the glaucoma subgroups was not statistically significant (p-value=0.588). However, a lower ECD were seen in Pseudoexfoliation Glaucoma (PXG) and Primary Angle Closure Glaucoma (PACG) subgroups. Conclusion: No significant difference was found in CCT and endothelial cell in cornea of patients with and without glaucoma. Normal Tension Glaucoma (NTG) patients have comparatively thinner corneas and OHTN patients had thicker corneas. There was no significant correlation established between ECD of Normal corneal vs. Glaucoma and its subgroups.

Corneal endothelial cell density and microvascular changes of retina and optic disc in autosomal dominant polycystic kidney disease.

Purpose:Vascular endothelial dysfunction in autosomal dominant polycystic kidney disease (ADPKD) may affect the retinal vascular parameters due to structural similarities of kidney and retina. We aimed to evaluate the microvascular changes of retina and optic disc and also corneal endothelial cell density in patients with ADPKD.Methods:Forty-six eyes of 23 patients with ADPKD (Group 1), and 46 eyes of 23 sex- and age-matched healthy controls (Group 2) were included in this cross-sectional study. Demographic and ophthalmic findings of participants were collected. Corneal endothelial cell density (CECD) measurements were obtained by noncontact specular microscopy. Foveal retinal thickness, peripapillary retinal nerve fiber layer (RNFL) thickness, vessel density in different sections of the retina and optic nerve head were analyzed by optical coherence tomography angiography.Results:The mean ages were 41 ± 11 years for Group 1 and 39 ± 10 years for Group 2 (P = 0.313). CECD values were significantly lower in group 1 when compared to group 2 (2653 ± 306 cells/mm2 and 2864 ± 244 cells/mm2, respectively, P < 0.001). The foveal retinal thickness and RNFL thickness were similar, but superior quadrant thickness of RNFL was significantly lower in Group 1 than Group 2 (126 ± 14 μm vs. 135 ± 15 μm, P = 0.003). In Group 1, whole image of optic disc radial peripapillary capillary densities were significantly lower compared to Group 2 (49.4 ± 2.04%, and 50.0 ± 2.2%, respectively, P = 0.043). There was no significant difference regarding superficial, deep retinal vessel densities, foveal avascular zone and flow areas between the groups (P > 0.05 for all).Conclusion:Lower CECD values and decreased superior quadrant RNFL thickness, and microvascular densities of optic disc were revealed in patients with ADPKD. Evaluation of CECD and retinal microvasculature may be helpful in the management of these patients.

Corneal In Vivo Laser-Scanning Confocal Microscopy Findings in Dry Eye Patients with Sjögren's Syndrome.

Purpose: To evaluate the changes in cornea in Sjögren’s syndrome (SS) with a novel confocal microscopy device. Methods: Twenty-three right eyes of patients with SS (23 women; mean age, 65.4 ± 11.4 years) and 13 right eyes of 13 age- and sex-matched control subjects (13 women; mean age, 68.8 ± 9.8 years) were studied. Furthermore, eight right eyes of patients with SS (8 women; mean age, 66.9 ± 9.6 years) were studied to evaluate the corneal microscopic alterations after the treatment with topical 3% diquafosol sodium eye drops. All cases had tear quantity, tear breakup time (BUT), ocular surface staining measurements, and corneal in vivo laser-scanning confocal microscopy examinations. The density and area of corneal epithelial cells (superficial, wing, and basal), density of corneal stromal cells (anterior, intermediate, and posterior), density and area of corneal endothelial cells, density and morphology of corneal sub-basal nerve plexus, density of corneal sub-basal inflammatory cells were also assessed. Results: The tear quantity, stability, and vital staining scores were significantly worse in patients with SS than in control subjects (p < 0.0001). Corneal superficial epithelial cell density was significantly lower in SS compared with control subjects (p < 0.0001). Corneal superficial epithelial cell area was significantly larger in SS compared with control subjects (p = 0.007). Corneal sub-basal nerve fiber density was lower in SS compared with control subjects (p < 0.0001). Morphological abnormality of nerve fibers was observed in SS patients. Corneal sub-basal inflammatory cell density was significantly higher in SS patients compared with control subjects (p < 0.0001). Furthermore, the mean corneal superficial epithelial cell density and area, inflammatory cell density, corneal sub-basal nerve fiber density, and morphological abnormality of nerve fibers, were improved with topical 3% diquafosol sodium treatment in the dry eye patients with SS (p < 0.05). Conclusions: The diagnostic modality using in vivo laser-scanning confocal microscopy was a useful method for the evaluation of the corneal cell density and area, nerve fiber density and morphology, and inflammatory cell density in patients with SS and also a useful tool in the assessment of treatment effect with topical 3% diquafosol sodium in the SS patients.

Correlations between corneal biomechanics and specular microscopy in patient with cataract

This study aimed to analyze the connection between corneal biomechanics (corneal hysteresis, CH) and endothelial cell density of cornea (mean endothelial cell density, MCD) in patients diagnosed with cataract. This retrospective, observational study was performed in the Ophthalmology Clinic of the University Emergency Hospital in Bucharest. Of 60 patients (120 eyes) with cataract, who were included in this study, we analyzed the CH values obtained using with the Ocular Response Analyzer (ORA) and the MCD values obtained using the specular microscopy. The study groups comprised both men and women with ages ranging from 45 to 63 years. Patients were divided into three study groups according to CH values. In each batch, the CH values obtained with the Ocular Response Analyzer (ORA) were correlated with age, gender and MCD, then the subgroups were compared. All the data gathered showed no correlation to be statistically significant regarding the biomechanical properties of the cornea and the corneal endothelial cell density in patients with cataract.

Outcomes of corneal transplantation using donor corneas retrieved from patients with chronic kidney disease

Purpose:To report the outcomes of corneal transplantation utilizing corneas retrieved from donors with chronic kidney disease (CKD).Methods:Outcomes of corneal transplantation (optical PK and EK) performed from Jan 2018 to Dec 2018 utilizing donor corneas retrieved from CKD patients was performed retrospectively.Results:Of the total of 233 donor corneas retrieved from CKD, 135 (57.9%) were utilized for transplantation after the routine screening protocol of the eye bank. Mean age of the donors was 56.2 ± 13.5 years. The mean endothelial cell density on specular microscopy of the donor corneas used for optical PK was 2685.7 ± 377.6 cells/mm2 (range, 2028–3448 cells/mm2) and for EK was 2731.7 ± 189.1 cells/mm2 (range, 2380–3194 cells/mm2). The overall primary graft failure rate was 5.1%. All grafts except 1, cleared in the PK group. In the EK group (6 DMEK and 16 DSAEK), 1 patient had a complete graft detachment and another 1 had a primary graft failure after DMEK.Conclusion:The donor corneas retrieved from chronic kidney disease patients are safe and suitable for optical keratoplasty provided they meet the criteria for transplantation.

Changes in corneal endothelial cell density in patients with primary open-angle glaucoma.

BACKGROUNDGlaucoma is a group of diseases characterized by a specific pattern of optic nerve neuropathy and retinopathy. Increasing evidence demonstrates glaucoma associated corneal endothelium loss. Direct-compression mechanism due to elevated intraocular pressure (IOP), cell toxicity after long term exposure to preservatives and glaucoma surgery have been reported to be the possible mechanism. Herein, we compare the specular endothelial microscopy in primary open-angle glaucoma (POAG) patients and healthy controls of the same age group to observe the corneal endothelium changes and the correlations to the mean IOP in a Chinese case control study.AIMTo investigate corneal endothelial cell density in Chinese patients with POAG.METHODSA case control study was performed on 60 eyes of 60 patients with POAG. Exclusion criteria included history of corneal diseases, intraocular diseases, contact lens use, ocular trauma or surgery (including intraocular surgery and laser treatment), congenital abnormalities or systemic diseases such as diabetes. Intraocular pressure was measured using Goldmann tonometry. Indirect specular microscopy (TOPCON SP-2000P) was performed on central corneas and endothelial images were acquired. Endothelial cell density, area and cell counts were analyzed.RESULTSEndothelial cell density was 2959 ± 236 cells/mm2 in healthy controls and 2757 ± 262 cells/mm2 in patients with POAG. The POAG eyes had significantly lower endothelial cell density compared to healthy control eyes (P < 0.001). In the POAG group, endothelial cell density was 2686 ± 233 cells/mm2 in the patients receiving medication and 2856 ± 272 cells/mm2 in the untreated subgroup. The eyes receiving medication had significantly lower endothelial cell density compared to untreated eyes. There was a negative correlation between cell density and mean IOP (r = −0.286, P = 0.004), positive correlation between the average cell area and mean IOP (r = 0.228, P = 0.022), maximum cell area and mean IOP (r = 0.218, P = 0.029) and minimum cell area and mean IOP (r = 0.290, P = 0.003). The percentage of hexagonal cells was not correlated with mean IOP.CONCLUSIONPatients with POAG have lower corneal endothelial cell density than healthy controls of the same age. This may be attributed to mechanical damage from elevated IOP and toxicity of glaucoma medications.

Corneal endothelial cell density and pterygium: a cross-sectional study.

To investigate the effects of pterygium on corneal endothelial cell density in patients with unilateral pterygium. We performed a cross-sectional analysis of data from patients with unilateral pterygium who were selected from September 1, 2015 to July 31, 2016 at Hospital de Base do Distrito Federal to assess the corneal endothelial cell density, coefficient of variation in the cell area, hexagonality, and corneal pa-chymetric results. In all patients, noncontact specular microscopy was performed in both eyes and a minimum endothelial cell count of 75 cells/mm2 was required for inclusion in the study. The contralateral eye served as the control. Sixty-one patients were included in the study. Twenty-nine (47.5%) patients were men and 32 (52.5%) were women (mean age, 50.84 ± 13.8). The percentage of pterygium that invaded the cornea ranged from 4.87% to 24.59% (median, 9.70% ± 4.99%). The mean corneal endothelial cell density (cells/mm) was lower in the pterygium eyes than in the controls (2451.83 ± 284.96 vs. 2549.95 ± 268.94, respectively; p=0.04). No differences in the mean coefficients of variation of cell size, hexagonality, and corneal pachymetric results were observed between the patients and controls. The Pearson correlation test showed a significant negative linear relationship between pterygium invasion and endothelial cell density [p<0.001, n=61, r=-0.553 (95% CI, -0.34 to -0.73)]. Compared with the contralateral eyes, those of patients with unilateral pterygium were associated with a decrease in corneal endothelial cell density.

Topical cyclosporine a for mustard gas induced ocular surface disorders.

Purpose:To evaluate the effectiveness of topical cyclosporine A 0.05% for treatment of mustard gas-induced ocular surface disorders with special attention to conjunctival goblet cell density in patients with severe dry eye.Methods:This prospective clinical study included 20 eyes of 20 patients previously exposed to mustard gas with dry eye syndrome unresponsive to artificial tears. Before and after treatment with topical cyclosporine A 0.05% twice daily for 3 months, subjects were evaluated for improvement in symptoms using the ocular surface disease index (OSDI) and signs by tear breakup time (TBUT), Schirmer test and measurement of superior bulbar conjunctival goblet cell density. Limbal stem cell deficiency (LSCD) and the degree of corneal squamous cell metaplasia were also assessed before and after treatment.Results:Before treatment, mean OSDI score, Schirmer test I value and mean TBUT were 42.8 ± 6.1, 4.2 ± 1.2 mm and 2.5 ± 1.3 s, respectively. After 3 months of treatment with topical cyclosporine A, these scores reached 36.4 ± 5.2, 5.8 ± 1.6 mm and 4.9 ± 2.1 s, respectively showing a statistically significant improvement (P < 0.001) in all parameters. Mean goblet cell density was 23.3 ± 17.1/high power field (hpf) at baseline which was significantly increased to 47.7 ± 16.1/hpf at the end of the study (P < 0.001). There was no improvement, however, in corneal conjunctivalization, LSCD and the degree of corneal squamous cell metaplasia based on impression cytology reports (P > 0.05).Conclusion:Treatment with topical cyclosporine A 0.05% in patients with severe dry eye due to mustard gas injury increases goblet cell density in the bulbar conjunctiva and improves symptoms of the disease.

Regional Alterations in Purkinje Cell Density in Patients with Autism

Neuropathological studies, using a variety of techniques, have reported a decrease in Purkinje cell (PC) density in the cerebellum in autism. We have used a systematic sampling technique that significantly reduces experimenter bias and variance to estimate PC densities in the postmortem brains of eight clinically well-documented individuals with autism, and eight age- and gender-matched controls. Four cerebellar regions were analyzed: a sensorimotor area comprised of hemispheric lobules IV–VI, crus I & II of the posterior lobe, and lobule X of the flocculonodular lobe. Overall PC density was thus estimated using data from all three cerebellar lobes and was found to be lower in the cases with autism as compared to controls, an effect that was most prominent in crus I and II (p<0.05). Lobule X demonstrated a trend towards lower PC density in only the males with autism (p = 0.05). Brain weight, a correlate of tissue volume, was found to significantly contribute to the lower lobule X PC density observed in males with autism, but not to the finding of lower PC density in crus I & II. Therefore, lower crus I & II PC density in autism is more likely due to a lower number of PCs. The PC density in lobule X was found to correlate with the ADI-R measure of the patient's use of social eye contact (R2 = −0.75, p = 0.012). These findings support the hypothesis that abnormal PC density may contribute to selected clinical features of the autism phenotype.

Endocrine cells in the oxyntic mucosa of the stomach in patients with irritable bowel syndrome.

To study the different endocrine cell types in the oxyntic mucosa of patients with irritable bowel syndrome (IBS). Seventy-six patients with IBS were included in the study (62 females and 14 males; mean age 32 years, range 18-55 years), of which 40 also fulfilled the Rome III criteria for functional dyspepsia (FDP). Of the entire IBS cohort, 26 had diarrhea as the predominant symptom (IBS-D), 21 had a mixture of diarrhea and constipation (IBS-M), and 29 had constipation as the predominant symptom (IBS-C). Forty-three age and sex-matched healthy volunteers without any gastrointestinal complaints served as controls. The patients were asked to complete the Birmingham IBS symptom questionnaire. Both the patients and controls underwent a standard gastroscopy, during which three biopsy samples were taken from the corpus. Sections from these biopsy samples were immunostained using the avidin-biotin complex (ABC) method, for ghrelin, serotonin, somatostatin and histamine. The densities of these cell types and immunoreactivity intensities were quantified using computerized image analysis with Olympus cellSens imaging software (version 1.7). The densities of the ghrelin cells in the control, IBS-total, IBS-D, IBS-M and IBS-C groups were 389 (320, 771), 359 (130, 966), 966 (529, 1154), 358 (120, 966) and 126 (0, 262) cells/mm(2), respectively. There was a significant difference between the tested groups (P < 0.0001). Dunn's multiple comparison test showed that the ghrelin cell density was significantly higher in IBS-D and lower in IBS-C than in the controls (P = 0.03 and 0.0008, respectively). The ghrelin cell density in patients with both IBS and FDP was 489 (130, 966), and in those with IBS only 490 (130, 956). There was no statistical significant difference between these 2 groups of patients (P = 0.9). The immunoreactivity intensity did not differ between any of the groups (P = 0.6). The diarrhea score of the Birmingham IBS symptom questionnaire was significantly positively correlated with ghrelin cell density (r = 0.65; P < 0.0001) and significantly inversely correlated with that of constipation (r = 90.69; P < 0.0001). The densities of the serotonin cells were 63 (51, 82), 51 (25, 115), 120 (69, 128), 74 (46, 123) and 40 (0, 46) cells/mm(2) in the control, IBS-total, IBS-D, IBS-M and IBS-C groups, respectively. A statistically significant difference was found between the tested groups (P < 0.0001). Posttest revealed that serotonin cell density was significantly higher in IBS-D and lower in IBS-C than in controls (P = 0.02 and 0.004, respectively), but did not differ in the IBS-total and IBS-M groups from that in controls (P = 0.5 and 0.4, respectively). The serotonin cell density in patients with both IBS and FDP was 62 (25, 115) and in those with IBS only 65 (25, 123). There was no statistically significant difference between these 2 groups of patients (P = 1). The immunoreactivity intensity of serotonin did not differ significantly between any of the groups (P = 0.0.9). The serotonin cell density was significantly positively correlated with the diarrhea score of the Birmingham IBS symptom questionnaire (r = 0.56; P < 0.0001) and significantly inversely correlated with that of constipation (r = 0.51; P < 0.0001). The densities of the somatostatin cells were 97 (72, 126), 72 (0, 206), 29 (0, 80), 46 (0, 103) and 206 (194, 314) cells/mm(2) in the control, IBS-total, IBS-D, IBS-M and IBS-C groups, respectively (Figures 7 and 8). There was a statistically significant difference between the controls and the IBS subgroups (P < 0.0001). The density of somatostatin cells was significantly lower in the IBS-D and IBS-M groups but higher in IBS-C patients than in the controls (P < 0.01, P = 0.02, and P = 0.0008, respectively). The somatostatin cell density in patients with both IBS and FDP was 86 (0-194), and in those with IBS only 110 (0-206). There was no statistically significant difference between these 2 groups of patients (P = 0.6). There was no significant difference in somatostatin immunoreactivity intensity between the controls. The diarrhea score of the Birmingham IBS symptom questionnaire was inversely correlated with somatostatin cell density (r = 0.38; P = 0.0007) and was positively correlated with that of constipation (r = 0.64; P < 0.0001). The finding of abnormal endocrine cells in the oxyntic mucosa shows that the endocrine cell disturbances in IBS are not restricted to the intestine. Furthermore, it appears that ghrelin, serotonin and somatostatin in the oxyntic mucosa of the stomach may play an important role in the changing stool habits in IBS through their effects on intestinal motility.