Superficial Epithelial Layers Research Articles

Soft-tissue keratocyst: report of 3 new cases from Brazil.

To describe the clinicopathological features of 3 new cases of soft-tissue keratocyst (STK) of the buccal mucosa from Brazil. Clinical data from 3 patients diagnosed with STK were obtained from the archives of the Laboratory of Oral Pathology at the Federal University of Rio de Janeiro- Brazil from 2020 to 2023. Two oral pathologists reviewed conventional hematoxylin and eosin-stained slides of each case. Immunohistochemical stainings for CK19, Bcl-2, CD138, D2-40, EMA, and Ki-67 were performed in all cases. Case 1 was a 58-year-old man with a 2-year history of painless swelling of the right buccal mucosa, measuring approximately 5cm. Case 2 was of a 44-year-old man with a painful swelling in the left buccal mucosa lasting 3 years with drainage to the oral cavity. Case 3 was of a 74-year-old woman with a painful swelling in the left buccal mucosa of unknown duration. Microscopic evaluation of all 3 cases revealed a cyst lined with a few cell layers composed of columnar palisading basal cells and a corrugated parakeratin surface. The capsule comprised fibrous connective tissue with variable amounts of blood vessels with hemorrhage, nerve bundles, fat tissue, striated muscle fibers, and the absence of pilosebaceous units. No recurrence was observed after complete surgical removal. All epithelial layers of the 3 cases expressed positivity for CK19 and CD138, the basal cells were positive for Bcl-2 and D2-40, and the superficial epithelial layer was positive for EMA. All cases demonstrated a low proliferation index by Ki-67. This study represents a series of 3 cases of STK that affected the posterior buccal mucosa of middle-aged patients from Brazil, sharing histopathological and immunohistochemical features with odontogenic keratocyst.

Pre-Descemet Corneal Dystrophy – Changes in Corneal Morphology in Confocal Microscopy

PurposeTo identify and describe the in-vivo microstructural changes in the cornea of patients with pre-Descemet’s membrane corneal dystrophy.MethodsTen patients (20 eyes) were examined in the Laser Microsurgery Centre in Warsaw. The ophthalmic examination, including visual acuity and slit lamp biomicroscopy, was carried out. Because of suspicion of corneal dystrophy, in vivo confocal microscopy of the cornea was also performed.ResultsBiomicroscopy revealed bilateral, dust-like fine opacities in the deep parts of the corneal stroma. Confocal microscopy showed pleomorphic structures (enlarged keratocytes) containing dense, hyperreflective, granular inclusions in the posterior stroma, next to the Descemet’s membrane. In 3 patients (6 eyes) punctate particles were seen extracellularly also in the mid stroma. Superficial and basal epithelial layers, anterior stroma and endothelium cells appeared normal.Conclusions1. In vivo confocal microscopy findings of cornea with pre-Descemet’s membrane dystrophy are characteristic. 2. In vivo confocal microscopy is a useful tool in diagnosing rare corneal dystrophies in cases where classical examination methods give inconclusive results.

Short beak and dwarfism syndrome among Pekin ducks: First detection, full genome sequencing, and immunohistochemical signals of novel goose parvovirus in tongue tissue.

Novel goose parvovirus (NGPV) is continuously threatening the global duck industry, as it causes short beak and dwarfism syndrome among different duck breeds. In this study, we investigated the viral pathogenesis in the tongue of affected ducks, as a new approach for deeper understanding of the syndrome. Seventy-three, 14- to 60-day-old commercial Pekin ducks were clinically examined. Thirty tissue pools of intestine and tongue (15 per tissue) were submitted for molecular identification. Clinical signs in the examined ducks were suggestive of parvovirus infection. All examined ducks had short beaks. Necrotic, swollen, and congested protruding tongues were recorded in adult ducks (37/73, 51%). Tongue protrusion without any marked congestion or swelling was observed in 20-day-old ducklings (13/73, 18%), and no tongue protrusion was observed in 15-day-old ducklings (23/73, 32%). Microscopically, the protruding tongues of adult ducks showed necrosis of the superficial epithelial layer with vacuolar degeneration. Glossitis was present in the nonprotruding tongues of young ducks, which was characterized by multifocal lymphoplasmacytic aggregates and edema in the propria submucosa. Immunohistochemical examination displayed parvovirus immunolabeling, mainly in the tongue propria submucosa. Based on polymerase chain reaction, goose parvovirus was detected in 9 out of 15 tongue sample pools (60%). Next-generation sequencing confirmed the presence of a variant goose parvovirus that is globally named NGPV and closely related to Chinese NGPV isolates. Novel insights are being gained from the study of NGPV pathogenesis in the tongue based on molecular and immunohistochemical identification.

Cytologic features of the bulbar conjunctiva in patients with primary open-angle glaucoma-associated dry eye disease

Purpose: To examine the features of the bulbar conjunctiva in patients who developed dry-eye disease (DED) after drug treatment for primary open-angle glaucoma (POAG). Methods: Impression cytology was performed by applying twice a strip of cellulose acetate filter to the ocular surface to remove the superficial epithelial layers of the temporal bulbar conjunctiva. The strips were removed with a peeling motion in a few seconds, and the samples were immediately fixed in 95% ethyl alcohol, stained with hematoxylin and eosin, mounted on glass slides and coverslipped for light microscopy. Squamous metaplasia was graded according to Nelson’s grading system on the basis of cell morphology, staining and integrity as well as the nucleus-to-cytoplasm ratio. This study included a case group of 80 patients (mean age, 63.8 +- 6.7 years) with POAG-associated DED, with the group being divided into four subgroups. Subgroups 1 and 2 were composed of 40 patients each, with glaucoma duration of less or more than 5 years, respectively. Subgroups a and b were composed of 40 patients each, with a number of topical ocular hypotensive drugs used equal to one or at least two, respectively. The control group was composed of 20 apparently healthy volunteers (mean age, 67.9 +- 8.9 years). All patients underwent a routine eye examination. Results: All patients with glaucoma had symptoms of DED with Ocular Surface Disease Index (OSDI) scores of at least 15. In subgroup 1, 60% had Nelson’s grade 1 and 40%, Nelson’s grade 2 squamous metaplasia. In subgroup 2, 10% had Nelson’s grade 1; 60%, Nelson’s grade 2 and 30%, Nelson’s grade 3 squamous metaplasia. In subgroup a, 20% had Nelson’s grade 1; 60%, Nelson’s grade 2 and 30%, Nelson’s grade 3 squamous metaplasia. In subgroup b, 10% had Nelson’s grade 1; 60%, Nelson’s grade 2 and 30%, Nelson’s grade 3 squamous metaplasia. Conclusion: Changes in the bulbar conjunctival epithelium corresponded to Nelson’s grade 2 or 3 squamous metaplasia in 80% of patients who developed DED after drug treatment for POAG. The severity of squamous metaplasia correlated with the duration of glaucoma and, consequently, longer use of hypotensive eye drops (r1 = 0.15, p1 = 0.02, p2 = 0.01). Findings of the current study and international guidelines argue for the use of the medications containing no preservatives or potentially toxic components in long-term therapy against glaucoma.

Safety Evaluation of Far-UV-C Irradiation to Epithelial Basal Cells in the Corneal Limbus.

Basal cells in the corneal limbus play an important role in the turnover cycle because they are the source of all cells that constitute the corneal epithelium. We examined the penetration depth of ultraviolet (UV) light in the corneal limbus and assessed the safety of Far-UV-C on stem cells in the basal area of the corneal limbus. Rats were irradiated with UV at peaks of 207, 222, 235, 254 and 311 nm while under anesthesia. The UV penetration depth in the rat corneal limbal epithelium was wavelength dependent: 311 nm UV-B and 254 nm UV-C reached the basal cells of the epithelium, and 235 nm radiation reached the middle area; however, 207 and 222 nm UV-C reached only the superficial layer of the epithelium. Porcine cornea, which is similar to the human eye in size and structure, were irradiated with 222 and 254 nm UV-C. As in rats, 222 nm UV-C reached only the superficial layer of the porcine corneal limbal epithelium. These results indicate that Far-UV-C, such as radiation of wavelengths of 207 and 222 nm, could not reach corneal epithelial stem cells, i.e. the cells remained intact. It is unlikely that the turnover of the corneal epithelium is obstructed or disrupted by exposure to Far-UV-C.

Irradiation Alters the Expression of MUC1, CD44 and Hyaluronan in Oral Mucosal Epithelium.

Purpose: It is well established that cancer cells exploit aberrant synthesis of mucin 1 (MUC1) and hyaluronan (HA) synthesis along with HA's physiological cell surface receptor CD44. However, their role in irradiated oral tissue has not been reported previously. We, therefore, aimed to study MUC1, CD44 and HA immunohistochemically in irradiated oral mucosa and their role in the long-term effects after radiotherapy. Materials and Methods: Oral mucosal biopsies were obtained from healthy subjects as controls and from patients after radiotherapy for head and neck cancer (irradiated group) during dental implant surgery.The presence of MUC1, CD44, and HA in oral mucosa was studied by immunohistochemical methods. The differences in the localization and intensity in the oral epithelium between control and irradiated tissue were analyzed. Results: The staining intensity of MUC1 was confined to the superficial epithelial layer, whereas HA and CD44 were found in the cell membranes in the epithelial basal and intermediate layers of control specimens. In irradiated epithelium, MUC1 staining was distributed throughout all the layers of the oral epithelium, with significant staining in the basal and intermediate layers. Accordingly, HA and CD44 staining extended to involve the superficial cells of the irradiated epithelium. The staining pattern of MUC1 and CD44 showed significant changes in irradiated samples. Conclusions: Our results showed that the staining intensities of MUC1, CD44, and HA were significantly elevated in irradiated tissue compared to controls. MUC1, CD44, and HA are important markers and take part in long-term changes in the oral mucosa after radiotherapy.

Origin, form and function of extraembryonic structures in teleost fishes.

Teleost eggs have evolved a highly derived early developmental pattern within vertebrates as a result of the meroblastic cleavage pattern, giving rise to a polar stratified architecture containing a large acellular yolk and a small cellular blastoderm on top. Besides the acellular yolk, the teleost-specific yolk syncytial layer (YSL) and the superficial epithelial enveloping layer are recognized as extraembryonic structures that play critical roles throughout embryonic development. They provide enriched microenvironments in which molecular feedback loops, cellular interactions and mechanical signals emerge to sculpt, among other things, embryonic patterning along the dorsoventral and left-right axes, mesendodermal specification and the execution of morphogenetic movements in the early embryo and during organogenesis. An emerging concept points to a critical role of extraembryonic structures in reinforcing early genetic and morphogenetic programmes in reciprocal coordination with the embryonic blastoderm, providing the necessary boundary conditions for development to proceed. In addition, the role of the enveloping cell layer in providing mechanical, osmotic and immunological protection during early stages of development, and the autonomous nutritional support provided by the yolk and YSL, have probably been key aspects that have enabled the massive radiation of teleosts to colonize every ecological niche on the Earth. This article is part of the theme issue 'Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom'.

Coring vs scrolls: Inclusion of the muscularis layer on tissue scrolls of the urinary bladder leads to inaccurate miRNA expression

Formalin fixation with paraffin embedding (FFPE) is a method used to preserve tissues. FFPE tissues can be sampled through tissue cores or scrolls. In the study of urothelial carcinomas (UC), the use of a biopsy needle to acquire cores has the advantage of sampling the targeted superficial epithelial cell layer. In contrast, tissue scrolls are collected using a microtome, which includes surrounding tissues such as the muscularis layer. While both methods allow sample collection to extract small molecules, such as miRNAs, there is a lack of literature comparing these methods. MiRNAs are non‐coding RNAs responsible for gene regulation, altered in cancer. UC is the most common malignant bladder tumor in dogs, with 20,000 cases annually. The diagnosis of UC is challenging due to the small sample size of biopsies acquired by endoscopy. This project aimed to compare miRNA expression between the FFPE collection methods in UC (n=3) and healthy bladder (n=1) from dogs to understand the effect of including tissues surrounding a lesion in scrolls. MiRNA was extracted using a commercial kit, and the expression was achieved through RT‐qPCR. Cores targeting the epithelia and the muscularis layer were compared to their corresponding scrolls. MiR‐214, miR‐152, miR‐181a, and miR‐39 were differentially expressed when comparing cores with scrolls and the muscularis layer, while miR‐1842 and miR‐143 were not. Comparison between methods within individual samples also showed differential expression for all miRNAs. These results show that the inclusion of the muscularis layer results in misleading miRNA expression and verifies cores as a more accurate technique in miRNA extraction and expression, supporting further miRNA biomarker research.

Protective effect of quercetin against corneal toxicities induced by cisplatin in male mice

The Aim of study was conducted to evaluate the potential protective role of quercetin against the corneal toxicity which caused by administration of cisplatin in male mice. Thirty-six adult male albino mice were divided into three equal groups; Group I control, Group II cisplatin mice injected as single dose I.P of cisplatin 10 mg/kg, and Group III mice received Q 75 mg/ kg with cisplatin 10 mg/ kg. The treatment continues for eight days then the tissues were collected for histopathological study. The histopathological changes in the cornea of eye were demonstrated in groups of mice received cisplatin. The changes represented by damages of superficial epithelial layer, vacuolar degeneration, hyperplasia, separation of stromal layer and sever damage of the stromal hemorrhage, and perforation of sclera, edema and inflammatory cells infiltration as well as separation of descement membrane from stroma layer. While these changes were diminished when the mice are treated with Q in the combination of cisplatin in comparison to treated cisplatin alone mice. Cisplatin induced structural and histopathological changes in the cornea of adult albino rat that could be ameliorated by concomitant treatment with quercetin.

Evaluation of the effect of multipoint intra-mucosal vaginal injection of a specific cross-linked hyaluronic acid for vulvovaginal atrophy: a prospective bi-centric pilot study

BackgroundVulvo-vaginal atrophy (VVA) is one of the common consequences of estrogen deficiency especially after the menopause. Several studies have assessed the effects of Hyaluronic acid (HA) on physical and sexual symptoms associated with VVA with promising results. However, most of these studies have focused on subjective assessment of symptom response to topically administered preparations. Nonetheless, HA is an endogenous molecule and it is logical that its effects are best realized if injected in the superficial epithelial layers. Desirial® is the first crosslinked HA that is administered by injection in the vaginal mucosa. The aim of this study was to explore the effect of multipoint vaginal intra-mucosal injections of specific cross-linked hyaluronic acid (DESIRIAL®, Laboratoires VIVACY) on several clinical and patient reported core outcomes.MethodsA cohort bi-centric pilot study. The chosen outcomes included change in vaginal mucosa thickness, biological markers for collagen formation, vaginal flora, vaginal pH, vaginal health index, vulvo-vaginal atrophy symptoms and sexual function 8 weeks post Desirial® injection. Patients’ satisfaction was also assessed using the patient global impression of improvement (PGI-I) scale.ResultsA total of 20 participants were recruited between 19/06/2017 and 05/07/2018. At the end of the study, there was no difference in the median total thickness of the vaginal mucosa or in procollagen I, III or Ki67 fluorescence. However, there was a statistically significant increase in COL1A1 and COL3A1 gene expression (p = 0.0002 and p = 0.0010 respectively). There was also a significant reduction in reported dyspareunia, vaginal dryness, vulvar pruritus, vaginal chafing and significant improvement in all female sexual function index dimensions. Based on PGI-I, 19 patients (95%) reported varying degrees of improvement where, 4 (20%) felt slightly better; 7 (35%) better and 8 (40%) much better.ConclusionsMulti-point vaginal intra-mucosal injections, of Desirial® (a crosslinked HA) was significantly associated with the expression of CoL1A1 and CoL3A1 suggesting stimulation of collagen formation. Furthermore, there was a significant reduction in VVA symptomatology and a significant improvement in patient satisfaction and sexual function scores. However, there was no demonstrable change in the total vaginal mucosal thickness.Study registration ID-RCB: 2016-A00124-47, Protocol code number: LOCAL/2016/PM-001.

Apical contacts stemming from incomplete delamination guide progenitor cell allocation through a dragging mechanism.

The developmental strategies used by progenitor cells to allow a safe journey from their induction place towards the site of terminal differentiation are still poorly understood. Here, we uncovered a mechanism of progenitor cell allocation that stems from an incomplete process of epithelial delamination that allows progenitors to coordinate their movement with adjacent extra-embryonic tissues. Progenitors of the zebrafish laterality organ originate from the superficial epithelial enveloping layer by an apical constriction process of cell delamination. During this process, progenitors retain long-lasting apical contacts that enable the epithelial layer to pull a subset of progenitors on their way to the vegetal pole. The remaining delaminated cells follow the movement of apically attached progenitors by a protrusion-dependent cell-cell contact mechanism, avoiding sequestration by the adjacent endoderm, ensuring their collective fate and allocation at the site of differentiation. Thus, we reveal that incomplete delamination serves as a cellular platform for coordinated tissue movements during development.

Vaginal collagen I and III changes after carbon dioxide laser application in postmenopausal women with the genitourinary syndrome: a pilot study

Objective The aim of the study was to evaluate the clinical response and collagen remodeling in the vaginal wall after three sessions of carbon dioxide (CO2) laser application. Methods Fourteen postmenopausal women with vulvovaginal atrophy, aged 45–65 years and sexually active, were evaluated with clinical questionnaires, gynecological examinations and histological techniques before and after 20 weeks of treatment (ClinicalTrials.gov NCT03939078). Treatment consisted of 3-monthly sessions of the CO2 laser. Clinical questionnaires included the Vaginal Health Index, the Female Sexual Function Index and the International Consultation on Incontinence Questionnaires Short Form. Biopsies were taken from the lateral vaginal wall at week 0 (left wall) and week 20 (right wall). Tissue samples were stained with hematoxylin and eosin, Periodic Acid-Schiff, Picrosirius Red Stain and Orcein dyes. Immunohistochemical study was used to quantify collagens I and III in the samples. Results The mean age was 54.4 ± 4.5 years, and the average time of amenorrhea was 7.6 ± 5.1 years. The Female Sexual Function Index and the Vaginal Health Index Score values increased while the International Consultation on Incontinence Questionnaire Short Form score decreased after the programmed treatment. There was no significant change in vaginal pH. Histological studies showed increases in the total and superficial epithelial cell layers, and type III collagen fibers (from 10.86 ± 7.66 to 16.87 ± 3.96, p < 0.05), and immunohistochemical studies confirmed the significant increase in collagen III. Conclusion Histological findings revealed epithelial atrophy reversal and collagen remodeling of the vaginal wall. Immunohistochemical analysis showed an increase in collagen type III fibers.

Distribution of Corneal TRPV1 and Its Association With Immune Cells During Homeostasis and Injury.

PurposeGiven the role of corneal sensory nerves during epithelial wound repair, we sought to examine the relationship between immune cells and polymodal nociceptors following corneal injury.MethodsYoung C57BL/6J mice received a 2 mm corneal epithelial injury. One week later, corneal wholemounts were immunostained using β-tubulin-488, TRPV1 (transient receptor potential ion channel subfamily V member-1, a nonselective cation channel) and immune cell (MHC-II, CD45 and CD68) antibodies. The sum length of TRPV1+ and TRPV1– nerve fibers, and their spatial association with immune cells, was quantified in intact and injured corneas.ResultsTRPV1+ nerves account for ∼40% of the nerve fiber length in the intact corneal epithelium and ∼80% in the stroma. In the superficial epithelial layers, TRPV1+ nerve terminal length was similar in injured and intact corneas. In intact corneas, the density (sum length) of basal epithelial TRPV1+ and TRPV1− nerve fibers was similar, however, in injured corneas, TRPV1+ nerve density was higher compared to TRPV1− nerves. The degree of physical association between TRPV1+ nerves and intraepithelial CD45+ MHC-II+ CD11c+ cells was similar in intact and injured corneas. Stromal leukocytes co-expressed TRPV1, which was partially localized to CD68+ lysosomes, and this expression pattern was lower in injured corneas.ConclusionsTRPV1+ nerves accounted for a higher proportion of corneal nerves after injury, which may provide insights into the pathophysiology of neuropathic pain following corneal trauma. The close interactions of TRPV1+ nerves with intraepithelial immune cells and expression of TRPV1 by stromal macrophages provide evidence of neuroimmune interactions in the cornea.

Histological Studies on Ageing Changes in the Cornea of Indian Buffaloes (Bubalus bubalis)

Background: Affections of eye commonly encountered in all the species of animals. If these not treated in time, the vision may be hampered, this may impair the physical ability, utility and productivity of animals leading to economic loss to the animal owners. Corneal dystrophies like basement membrane dystrophy, stromal dystrophy, endothelial dystrophy, corneal erosions and corneal ulcerations etc., seen in the aged buffaloes. The current study was carried out to establish basic data and to provide authentic information to the clinicians on ageing changes in eyes of buffaloes.Methods: The study on age related changes in the cornea was conducted on 63 eye balls of locally available buffaloes of different breeds in and around Proddatur. These buffaloes were categorized into 3 groups based on their age i.e., group I (1-5 yrs), group II (6-10 yrs) and group III (11 yrs and above) buffaloes. The eyeballs were isolated and fixed with Davidson’s fluid. The paraffin sections of cornea were subjected for routine histological study.Result: The mean total thickness (µm) of cornea was increased from group I to III animals and it was 747.22±13.57, 853.33±11.05 and 897.22±8.06. Hence, The mean thickness (µm) of all five layers of cornea increased with advancement of age. Superficial corneal epithelial layer was made up of non keratinized stratified squamous epithelium with 6 to 7 layers at early age and 12-14 layers of cells in adult animals. The mean thickness (µm) of corneal epithelium in group I to III animals was 117.04±3.37, 122.59±2.23 and 133.71±3.55 respectively. Bowman’s membrane of cornea was measured 12.97±0.86, 17.22±0.55 and 18.71±0.68 in group I, II and III animals respectively. The mean thickness (µm) of corneal stroma in group I to III animals was 600.75±10.33, 672.22±10.46 and 690.00±5.32 respectively. This was mainly due to progressive increase in number of collagenous lamellae. The keratocytes became thin in old animals. Thickness of posterior band zone (PBZ) of Descemet’s membrane was increased with advancement of age. The mean thickness (µm) of Descemet’s membrane and endothelium together was in group I, II and III animals was 16.48±0.91, 41.29±2.25 and 54.82±1.44 respectively. The corneal endothelial cells were enlarged and increased in size in old buffaloes. The age related pigmentation was also noticed in epithelium and stroma of cornea in animals of group III. Density of sub basal nerve plexuses between the stroma and the corneal epithelium was not altered with advancement of age.

A comparative study of the clinical andhistopathological features of normal labial mucosa and redundant tissue in double lip

Introduction: double lip is one of lip abnormalities that may be congenital or acquired. It consists of a fold of excess tissue on the mucosal side of the lip. It causes functional and aesthetic problems to affected patientsAim of the work : The present study was performed to compare clinically and histopathologically between redundant tissue in double lip and normal labial mucosa.Subjects and methods: Seventeen patients with congenital non syndromic double lip were selected. Redundant tissues were removed surgically together with part of adjacent normal mucosa as part of the surgical manipulation. Hematoxylin and eosin staining was done to compare both tissues microscopically. Expression of ki-67 and p53 was evaluated in both tissue by immunohistochemistryResults: Clinically, redundant tissue appeared as thick extra fold with normal mucosal color and soft consistency analogous to normal labial mucosa. This redundant tissue was removed surgically without any complications. Histopathologically, normal labial mucosa revealed non keratinized stratified squamous epithelium and delicate underlying connective tissue. Excess tissue exhibited parakeratosis, acanthosis, elongation of rete pegs. The underlying connective tissue was fibrotic with inflammatory cell infiltration. In normal labial mucosa, ki-67 was detected only in parabasal layer of epithelium whereas it was localized in parabasal and superficial layers of epithelium in redundant tissue. No expression of p53 was detected in both normal and excess tissue. For both ki-67 and p53, the difference in expression between normal and redundant tissue was significant.Conclusion : Redundant tissue is a benign hyperplastic tissue that require surgical removal. So surgical excision here is the treatment of choice when excess tissue causes functional and aesthetic problems. Moreover, the high proliferation of redundant tissue may necessitate the surgical excision of this tissue.

Effect of Progestin Usage on the Interpretation of Cervical High-grade Squamous Intraepithelial Lesion

Progestin usage can alter the histologic and cytologic features of HSIL, which may potentially lead to the under-diagnosis of this precancerous lesion. The objective of the present study was to assess the relationship between the usage of progesterone-based contraceptives and the cytologic features of high-grade squamous intraepithelial lesion (HSIL) of the cervix. In this case-control study, we assembled 46 cases of cervical HSIL (CIN 3), including samples from 26 patients with known history of progestin usage (study group) and 20 samples from patients with no history of any exogenous hormone usage (control group). Cell image analysis was performed on all samples using a proprietary software. Immunohistochemical studies for Ki67, p16, estrogen receptor and progesterone receptor were performed on all cases, as was RNA in situ hybridization for HPV subtypes 16 and 18. Compared with the control group cases, the average nuclear size (21.5±1.80 μm) and the nuclear: cytoplasmic ratio (0.28±0.015) of HSILs in the study group cases were significantly smaller (P=0.026) and reduced (P=0.005), respectively. In addition, the study group cases showed reduced nuclear atypia and pleomorphism and significantly reduced mitotic figures (1.74±1.86/mm in study group versus 5.94±1.3/mm in control group, P<0.0001). The later was likely a consequence of the significantly reduced mitotic figures in the superficial and middle epithelial layers of the study group cases compared with their control group counterparts (0.73±0.88/mm vs. 7.3±2.6/mm, P<0.0001). Progestin usage has no discernible effects on p16 immunoreactivity, Ki67 proliferative index, hormone receptor expression, and HPV RNA levels of HSIL lesions. Being aware of progestin induced morphologic changes on those cervical precancerous lesions will be ultimately improve patient care quality in our daily practice.

Multimodal imaging of pre-Descemet corneal dystrophy.

The aim of this study was to assess structural and histological changes associated with pre-Descemet corneal dystrophy with multimodal in vivo imaging. Retrospective case series including eight corneas from four unrelated male patients with pre-Descemet corneal dystrophy characterized by the presence of punctiform gray opacities located just anterior to the Descemet membrane at slit-lamp examination of both eyes. In vivo confocal microscopy images were obtained in the central, paracentral, and peripheral corneal zones from the superficial epithelial cell layer down to the corneal endothelium in both eyes. Spectral domain optical coherence tomography scans (central and limbal zones) and mapping of both corneas were acquired. Diffuse small extracellular stromal deposits, presence of enlarged hyperreflective keratocytes in the posterior stroma with either hyperreflective or hyporeflective intracellular dots, and presence of activated keratocytes in the very anterior stroma were observed in all corneas with in vivo confocal microscopy. Spectral domain optical coherence tomography scans showed a hyperreflective line anterior to Descemet's membrane running from limbus to limbus and associated with a second thinner hyperreflective line just beneath Bowman's layer. Fine hyperreflective particles were observed in the posterior, mid, and anterior stroma on optical coherence tomography scans. The clinical presentation and structural anomalies found in isolated sporadic pre-Descemet corneal dystrophy are in favor of a degenerative process affecting corneal keratocytes with no epithelial or endothelial involvement. The maximum damage is found just anterior to the Descemet membrane resulting in pre-Descemet membrane location of stromal opacities. Multimodal imaging of cornea reveals that the disorder affects the whole stroma and it permits better understanding of pre-Descemet corneal dystrophy pathophysiology together with ascertained diagnosis.

Amniotic Membrane Transplant for Bullous Keratopathy: Confocal Microscopy & Anterior Segment Optical Coherence Tomography

ABSTRACTPurpose: To assess amniotic membrane retention after amniotic membrane transplant in bullous keratopathy patients and whether there were any corneal structural changes that may hinder further penetrating keratoplastyMethods: A retrospective study including 22 patients who have undergone amniotic membrane transplant from 1 Jan 1998 till 30 Jun 2016. Confocal microscopy and anterior segment optical coherence tomography (ASOCT) were performed to assess the retention of amniotic membrane and to detect any corneal structural changes. The comparison was made with 5 controls who had bullous keratopathy awaiting endothelial keratoplasty.Results: Patients had a mean follow-up of 61 ± 33.7 months. Pain reduction was significant (p < .001) although it did not significantly correlate with the regularity of the superficial, intermediate or basal epithelial layers, nor with the retention of the amniotic membrane. No long-term structural changes that may hinder future penetrating keratoplasty were detected.Conclusion: This procedure is a safe and effective long-term treatment for symptomatic bullous keratopathy patients.

Detection of intrinsic cholinergic system in the human lacrimal drainage system: evidence and potential implications.

To investigate the presence and distribution of epithelial and non-epithelial cholinergic system and cholinergic brush cells in the human lacrimal drainage system. The study was performed on fresh frozen human cadaveric samples ofthe lacrimal drainage system. Immunohistochemistry was performed for assessing the presence and distribution of cholinergic brush cell proteins-villin, acetylcholine synthesizing enzyme, choline acetyltransferase (ChAT); vesicular acetylcholine transporter (VAChT); components of canonical taste transduction signaling cascade, phospholipase C β2 (PLCβ2), and transient receptor potential cation channel, subfamily M, and member 5 (TRPM5). In addition, immunoreactivity to carbonic anhydrase 4 (CA4) was assessed. The immunoreactivity was scored as positive or negative and the distribution patterns in the canaliculi, lacrimal sac, and nasolacrimal duct were investigated. In addition, ultrastructural analysis was performed to ascertain the presence of brush cells by means of scanning electron microscopy (SEM). Villin revealed immunoreactivity in the superficial epithelial cells of lacrimal sac and nasolacrimal ducts. Positive immunoreactivity was also found for ChAT, VAChT, TRPM5, and PLCβ2. ChAT expression was limited to the superficial epithelial layers of the lacrimal sac epithelium. TRPM5 and PLCβ2 were expressed on the cell membranes, cytoplasm, and basolateral surfaces of the lacrimal sac epithelium and also showed strong expression in the submucosal glandular acinar cells. VAChT showed strong expression in the canaliculus and lacrimal sac and was expressed on the surface of the superficial epithelial cells and the submucosal glandular acinar cells and lining of the blood vessels. There was a uniformly negative immunoreactivity for CA4. SEM revealed single epithelial cells with dense tuft of rigid apical microvilli in the lacrimal sac and nasolacrimal ducts. This study provides a proof of principle for the presence of an intrinsic epithelial cholinergic mechanism in the lacrimal drainage system.

PS02.025: CHARACTERISTICS OF SUPERFICIAL BASALOID SQUAMOUS CELL CARCINOMA TREATED BY ENDOSCOPIC RESECTION

Abstract Background Basaloid squamous cell carcinoma (BSCC) is a rare esophageal carcinoma. And the characteristics especially in early stage is unknown. Methods The aim of this study is to clarify the characteristics of superficial BSCC. Thirteen patients (9 males and 4 females) with BSCC treated by endoscopic resection (12 ESD and 1 EMRC) from January 2000 to March 2017 were enrolled in this retrospective study. The median age was 67 (57–83). The median follow-up period was 24 (12–115) months. T1a-EP/LPM and T1a-MM/T1b-SM1 without lymph duct invasion (LDI) was followed up. T1a-MM/T1b-SM1 with LDI or T1b-SM2 were treated by additional treatment (AT). Results 1. En-bloc resection and R0 resection rate was 100% and 100%, respectively. 2. Macroscopic type; 0-IIa, 0-IIb and 0-IIc was 4, 1 and 8, respectively. 3. The median tumor diameter was 27 (2–62) mm. 4. Invasion depth; T1a-LPM, T1a-MM/T1b-SM1, and T1b-SM2 was 3, 5 and 5, respectively. 5. Histological characteristics; All BSCC were covered by SCC or non-neoplastic squamous epithelium. 6. Endoscopic characteristics; SMT like appearance was found in 46% (6/13). Yellowish nodules were observed in 23% (3/13). This finding was observed, when BSCC exist superficial epithelial layer covered by thin non-neoplastic epithelium. 7. Prognosis A: T1a-LPM (N = 3). Two patients are alive without recurrence, and 1 patient died of other disease. B: T1a-MM/SM1 (N = 5). Two of 3 patients without LDI were followed up and alive. Remaining one patient wanted to be treated by CRT, and alive. Two patients had LVI, however they were followed up without AT because of patient's hope. One is alive, and another died of other disease. C: T1b-SM2 (N = 5). Two and one patients were treated by surgery and chemotherapy, respectively, and alive without recurrence. Remaining two patients were followed up without AT. One of 2 patients are alive without recurrence. Another patient dead of other disease. Conclusion 1. All of BSCC were covered by SCC or non-neoplastic epithelium. 2. SMT like appearance and yellowish nodules under squamous cell epithelium were the characteristic endoscopic findings. 3. 40% of T1aMM/T1bSM1 had LDI. It's higher than that of SCC. Therefore, En-bloc resection is necessary for the detailed histological examination. Disclosure All authors have declared no conflicts of interest.