Basal Epithelial Cell Layer Research Articles

Meibomian Gland Probing Stimulates a Proliferative Epithelial Response Resulting in Duct Regeneration.

To demonstrate that the meibomian gland ductal basement membrane and basal epithelial cell layer are in continuity with and may derive from lid margin orifice-associated rete ridge epithelial/basement membrane structures (OARREBS) and to characterize changes in the distal duct microanatomy after meibomian gland probing (MGP) using in vivo confocal microscopy (IVCM). Pre/post-MGP IVCM examinations were performed on upper lids. Thirty-six identical glands from 20 lids of 16 patients (49.24 ±17.11 y/o with 13:3 F:M) were identified, analyzed, and compared to control cases. Statistical analyses were performed using ImageJ software and IBM SPSS version 27. All MGPs were performed within 12 weeks of the initial examination. Post-MGP follow-up exams occurred at 5.03 ±4.48 months. Post-MGP images showed more superficially organized OARREBS with accelerated and more superficial basement membrane formation, and an average increase of 32.2%, 25.4%, 32.04%, 77.7%, and 81.3% in duct wall epithelial cell layers (DWECL) (p < 0.001, compared to control (CTC) p < 0.001), distal duct wall thickness (DWT) (p < 0.001, CTC p < 0.001), proximal DWT (p < 0.001, CTC p < 0.001), distal lumen area (p < 0.001, CTC p = 0.037), and proximal lumen area (p < 0.001, CTC p = 0.007), respectively. The increase in the distal DWT and lumen area correlated with the months of follow-up (p = 0.004 and p = 0.010, respectively). Immediate post-MGP imaging revealed the probe track confined to the ductal epithelial compartment. MGP appears to stimulate a proliferative epithelial response characterized by an accelerated more superficial formation of ductal basement membrane with increased DWECL as well as DWT and lumen area at two separate duct foci. These findings suggest activation of lid margin meibomian gland precursor cells and confirm that MGP stimulates an epithelial regenerative phenomenon, not a fibrotic one.

LBSUN294 Insulin-induced Abdominal Acanthosis Nigricans From High Doses Of Insulin U-500

Abstract Introduction Dermatological complications associated with insulin therapy include lipoatrophy, lipohypertrophy, and acanthosis nigricans. We have a rare case of severe acanthosis nigricans from repeated insulin injection. Case A 63-year-old man with diabetes mellitus type 2 presented with an elevated blood glucose of 584mg/dL. He reported "high" glucose levels, despite adherence to his insulin regimen. He only injected insulin in his abdomen. Over 20 years, his insulin doses increased from Lantus 65 units BID and Humalog 35 units TID to Lantus units 110 BID and Humalog 65 units TID. It was later changed to U-500 at 160 units TID. His Hba1c ranged from 10.7% to 15.4%, most recently 13.6%. Physical examination revealed hyperpigmented and thickened plaques, localized to two large areas lateral to his umbilicus. During his hospital stay, insulin was administered on his arms and thighs. Glucose levels improved on Levemir 55 units daily and Lispro 20 units TID. He was discharged on Insulin U-500 40 units TID with instructions to avoid injecting on his abdomen and to rotate injection sites. Discussion Acanthosis nigricans is associated with insulin resistance or hyperinsulinemia. Insulin has a similar structure to insulin-like growth factor (IGF-1). At higher insulin concentrations, it can bind to IGF-1 receptors on keratinocytes and fibroblasts, leading to skin proliferation. Other suggested mechanisms include the involvement of fibroblast and epidermal growth factor. Histopathological findings show hyperkeratosis, papillomatosis, and thickened dermis. Immunochemical findings show positive IGF-1 antibodies in basal epithelial and prickle cell layers. In patients with poor glycemic control and increasing doses of insulin, it is important to assess for insulin-related skin changes. Besides routine visual inspection, skin thickening can be assessed by stroking the skin side-to-side in a sweeping motion instead of palpation. Conclusion Our patient with poorly type 2 diabetes required escalating doses of insulin over the years. He developed severe acanthosis nigricans on his abdomen leading to poor absorption of insulin. This case highlights the importance of routine visual inspection and tactile examination of insulin injection sites for skin changes, as well as repeated counseling regarding insulin site rotation.

A new diagnostic paradigm for Erythromelanosis Follicularis Faciei et Colli: Dermoscopic and reflectance confocal microscopic features

Introduction: Erythromelanosis Follicularis Faciei et Colli is a rare variant of Kerastosis pilaris, characterized by a clinical triad of erythema, hyperpigmentation and follicular papules involving bilaterally cheeks, preauricular and maxillary areas of young male or, more rarely, female patients Case presentation: A 21-year-old Caucasian woman presented symmetrical patches of hyperpigmentation, erythema with microscopic telangiectasias and tiny follicular papules involving cheeks, preauricular and maxillary areas, forehead, pinna and neck since childhood. Lesions were asymptomatic but caused profound cosmetic disfigurement and had a negative impact on her social life. The patient underwent a non-invasive diagnostic procedure with epiluminescence and confocal microscopy. Dermoscopic examination revealed prominent yellowwhitish follicular keratotic plugging over a reddish background with short fine telangiectasia in the perifollicular and interfollicular areas. Furthermore, RCM features displayed the presence of enlarged the presence of enlarged follicular openings, absence of demodex mites within the follicles and slight infundibular hyperkeratosis. At the level of the epidermal-dermal junction hyperpigmentation of the epithelial basal cell layer and dilation of dermal capillaries inside dermal papillae were observed, correlating to typical histological features of EFFC. Our patient benefited from the topical use of tretinoin 0.05% cream, with attenuation of the instrumental pictures. Conclusion: Characteristic plugging over a reddish background and morphological features revealed by RCM may be helpful for assessment and monitoring in patients with EFFC. Starting from the observation of this case we would like to underline the importance of a noninvasive diagnostic procedure and an effective therapy for such a rare condition. Keywords: Erythromelanosis Follicularis Faciei et Colli; Erythromelanosis Follicularis Faciei diagnosis; Dermoscopy; In vivo reflectance confocal microscopy.

Novel Insight of Cytokeratin 14 as a Biomarker in Diagnosing Bladder Cancer

Bladder cancer is the ninth most common type of cancer occurring in men. Screening for bladder cancer is often difficult due to limited proper modality and symptoms that is mild and unspecific. Leaving it untreated leads to greater morbidity and mortality. Therefore, finding a simple biomarker for screening purpose is necessary. Cytokeratin (CK) has been studied extensively as cancer biomarker. Detectable soluble CK was found in body fluid, including urine, and histological specimen using immunoassays. The level of its expression is mainly affected by cell proliferation, apoptosis, and cellular stress. CK 14, which is type I CK, was detected in the basal layer of epithelial cells and specific for squamous type of epithelial cell. CK 14 analysis in bladder cancer has revealed numerous functions such as predicting the development of bladder cancer, differentiating cancer origins, stratifying bladder cancer patients based on disease severity, and predicting bladder cancer recurrence. Its potential to become excellent biomarker makes it subject of interest in research development.

Transcriptome analysis identifies the differentially expressed genes related to the stemness of limbal stem cells in mice.

Limbal stem cells (LSCs) reside in the basal layer of limbal epithelial cells (LECs). They are crucial for maintenance of corneal epithelium homeostasis and corneal wound healing. Their stemness is determined by their gene expression pattern. Despite of several positive identifiers have been reported, the unique biomarker for LSCs still remain elusive. Differentially expressed genes (DEGs) between stem cells and differentiated cells affect the fate of stem cells via specific signaling pathway. In order to understand the DEGs in the LSCs, RNA-seq was firstly conducted using a mouse model. A total of 1907 up-regulated DEGs and 395 down-regulated DEGs were identified in the limbus (L) compared to central cornea (CC) and conjunctiva (Cj). Reliability of the expression of genes from RNA-seq analysis was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and immunofluorescence staining. The expression pattern of putative biomarkers was considered to be age-related. In up-regulated DEGs GO analysis, 570 gene ontology (GO) terms were significantly enriched. Five groups of genes related with biological processes from these significantly enriched GO terms comprised ionic transport, regulation of tissue development, muscle contraction, visual perception, and cell adhesion, which were clustered as a weighted similar network. Whereas, in down-regulated DEGs GO analysis, 61 GO terms were significantly enriched and only one group of ATP biosynthesis and metabolic process were clustered. Furthermore, we identified 55 signaling pathways by the Kyoto Encyclopedia of Genes and Genomes (KEGG) database based on up-regulated genes and 14 KEGG pathways based on down-regulated genes. In this study, we provide a landscape of the expression of putative LSCs biomarkers and stemness-related signaling pathways in a mouse model. Our findings could aid in the identification of LSC niche factors that may be related to the stemness of the LSCs.

Immunohistochemical profile of the anti-apoptosis, apoptosis and proliferation markers Bcl-2, caspase-3, p53, and Ki-67 in botryoid odontogenic cysts compared to lateral periodontal cysts and gingival cysts of the adult

ABSTRACT We investigated the differences in growth and rates of recurrence of the botryoid odontogenic cyst (BOC) and the less aggressive lateral periodontal cyst (LPC) and gingival cyst of the adult (GCA). We compared the immunohistochemical expression of selected biomarkers of apoptosis and proliferation and of regulators of their activity. Sections from archival paraffin blocks of 15 BOCs, six GCAs, six LPCs, and three odontogenic keratocysts (OKCs) were processed for immunohistochemical localization of Bcl-2, caspase-3, p53 and Ki-67. Labeled and unlabeled epithelial cells were counted and differences in the mean labeling index (LI) were evaluated statistically. The only significant differences in LI were for the anti-apoptotic marker, Bcl-2; the hierarchy was BOC > OKC > LPC > GCA. In two BOCs, 97% of the cells, and in all OKCs, all of the basal cells were labeled with Bcl-2. Otherwise, cells labeled with Bcl-2, p53 and caspase-3 were scattered among the basal and intermediate epithelial cell layers. Ki-67 labeled almost exclusively basal cells in the BOCs, LPCs and GCAs, and both basal and intermediate layer cells in the OKCs. Our findings suggest that while there was no significant difference in replicative potential of the GCAs, LPCs and BOCs, factors that influence apoptosis may be partially responsible for the more aggressive behavior of BOCs and OKCs.

Silver Sulfadiazine's Effect on Keratin-19 Expression as Stem Cell Marker in Burn Wound Healing.

BackgroundBurn wounds are one of the causes of cutaneous injury that involve both epidermal and dermal layers of skin. Silver sulfadiazine (SSD) has been widely used to treat burn wounds, however recent studies have found the treatment to have some drawbacks, such as cellular toxicity effects. Cutaneous wound regeneration is known to start from the basal layer of the epidermal epithelial cells, which are enriched with highly proliferative cells. Keratin-19 (K19) is one of the epidermal stem cell biomarkers found in the skin. This study aims to explore the expression of K19 in burn wound tissue and to investigate the effect of SSD on its expression.MethodsWe created a burn wound model in Sprague Dawley rats and randomly divided them into control and SSD groups. Wound closure was evaluated (visitrak) overtime series followed by histological evaluation of K19 expression in the wound tissue (immunohistochemistry staining).ResultsOur model successfully represents full-thickness damage caused by a burn wound. The SSD group showed a faster reduction of wound surface area (wound closure) compared to the control group with the peak at day 18 post wounding (p < 0.05). K19 expression was found in both groups and was distributed on epidermal layers, hair follicles and dermis of granulation tissue showing similar patterns.ConclusionTopical application of SSD on burn wounds showed superiority in wound closure and is likely to have no harmful effect on epidermal stem cells. However, further study is required to investigate the effect of silver species on cell viability and toxicity effects during long term treatment.

Post-LASIK Ectasia Versus Keratoconus: An In Vivo Confocal Microscopy Study

To examine and compare corneal cellular and subbasal nerve (SBN) characteristics in post-laser-assisted in situ keratomileusis ectasia (PLE) corneas, normal post-laser-assisted in situ keratomileusis corneas (PLC), keratoconus (KC) corneas, and normal virgin corneas (NC). In this cross-sectional comparative study, 18 PLE eyes of 11 patients, 18 PLC of 15 cases, 32 KC eyes of 32 patients, and 29 NC of 29 subjects were assessed using in vivo confocal microscopy. The density of the basal epithelial cell (BEC), anterior keratocyte, posterior keratocyte, and endothelial cell layers, as well as the characteristics of SBN fibers, was compared between the 4 groups. The density of the BEC and anterior and posterior keratocyte layers was significantly lower in KC compared with NC (-650 ± 190, P = 0.013; -181 ± 39, P < 0.001; and -36 ± 11, P = 0.021, respectively). However, there was no significant difference between PLE and PLC regarding these parameters (all Ps ≥ 0.6). Mean SBN parameters, including central corneal nerve branch density, nerve fiber length, total branch density, and nerve fiber area, were significantly lower in KC compared with NC and in PLE compared with PLC (all Ps ≤ 0.021). The pathophysiology of PLE seems to differ from KC. Apparent changes in the BEC and anterior and posterior keratocytes associated with KC were not observed in PLE. However, SBNs seem to be involved in both conditions.

Expression of keratins 8, 18, and 19 in epithelia of atrophic oral lichen planus.

Keratins form intermediate filaments of the cytoskeleton in keratinocytes and have roles in cell structure, signaling, intracellular transport, and cell death. Oral lichen planus (OLP) is an oral inflammatory disease with derangements in basal keratinocytes and disruption of the basal membrane. Here, we focused on epithelial expression of keratins 8, 18, and 19 because these proteins are known to modulate cell death. Biopsies were taken from buccal oral mucosa of persons with normal oral mucosa (n=10) or atrophic OLP (n=10). Cultured normal oral keratinocytes (n=4) showed expression of mRNA and protein for keratins 8, 18, and 19. Immunohistochemistry showed consistent staining for keratins 8 and 18 in basal keratinocytes of normal oral mucosa. In OLP, staining for keratin (K)8 was mostly negative and staining for K18 was weak. Keratin 19 was expressed irregularly in most biopsies of normal oral mucosa and not at all in OLP. Several mononuclear leukocytes in the cellular infiltrate showed membrane staining for K8 and K18. Positive staining for K16 confirmed partial collapse of the basal cell layer in OLP. The basal cell niche in OLP therefore appeared to be partly populated with keratinocytes demonstrating a higher degree of differentiation (K8- K18- K19- K16+ ); consequently, such areas may be more susceptible to the action of cell death factors released from the cell infiltrate as a result of lacking the protective, normal keratin present in the basal epithelial cell layer of normal oral mucosa.

Keratin 5/14‑mediated cell differentiation and transformation are regulated by TAp63 and Notch‑1 in oral squamous cell carcinoma‑derived cells.

Keratins5/14 (K5/14) are intermediate filament proteins expressed in the basal layer of stratified epithelial cells and are known targets of p63. Previous research in our laboratory showed that upon K5/14 downregulation in oral squamous cell carcinoma (OSCC)‑derived cells, there was an increase in intracellular Notch‑1 levels and differentiation markers such as involucrin, keratin1 and a decrease in tumorigenic potential invivo. However, the molecules involved in the K14 regulated cell differentiation and transformation are not known to date. In order to understand the possible role of TAp63, we downregulated TAp63 in a K14‑knockdown background. We observed that there was a decrease in the expression of Notch‑1. Expression levels of differentiation markers such as involucrin, K1, loricrin and filaggrin were also decreased. Furthermore, TAp63 downregulation led to an increase in invasion, migration and invivo tumorigenic potential of these cells. We observed a decrease in β‑catenin signaling in K14‑downregulated cells. Notably, when TAp63 was downregulated in K14‑knockdown cells, there was increase in non‑phospho β‑catenin levels. Hence, this study indicates that TAp63 plays an important role in K14‑downregulated cells possibly by regulating the Notch‑1 expression. K14 regulates the expression of TAp63 which in turn regulates expression of Notch‑1. The present study is a step forward in our quest to understand the functional significance of molecules that regulate the process of differentiation and tumorigenesis in stratified epithelial cells.

Histological and microRNA Signatures of Corneal Epithelium in Keratoconus.

To illustrate the histopathology of keratoconic corneal epithelia and its micro-ribonucleic acid (miRNA) regulation. Corneal epithelia were collected from 27 patients with keratoconus and 26 normal patients after surgery or by impression cytology. The miRNA profile was determined using miRNA microarray. The biological roles of miRNA target genes were delineated by gene ontology and pathway analyses. The expressions of significant miRNAs were validated using TaqMan polymerase chain reaction (PCR), whereas protein localization and expression of the miRNA target genes were examined by immunofluorescence and immunoblotting analyses. Histological assessment showed that corneal epithelia in patients with keratoconus were thinner with loosely packed cells compared to normal patients. Microarray analysis revealed that 12 miRNAs were significantly downregulated in keratoconic corneal epithelia. Gene ontology analysis demonstrated that the predicted miRNA target genes participated in cell junction, cell division, and motor activity, whereas pathway analysis highlighted the involvement of syndecan-mediated signaling pathway. TaqMan PCR validated the altered expression of six miRNAs in corneal epithelia from surgery (hsa-miR-151a-3p, hsa-miR-138-5p, hsa-miR-146b-5p, hsa-miR-194-5p, hsa-miR-28-5p, and hsa-miR-181a-2-3p) and four miRNAs in squamous corneal epithelial samples collected from impression cytology (hsa-miR-151a-3p, hsa-miR-195-5p, hsa-miR-185-5p, and hsa-miR-194-5p). In addition, higher S100A2 expression was found in the epithelial basal cell layer of keratoconic corneal epithelia. The miRNA and histological analyses in this study demonstrated structural and biological changes in keratoconic corneal epithelia, broadening the understanding of keratoconus pathology. In addition, impression cytology is useful to collect corneal epithelial tissues for gene expression analysis. [J Refract Surg. 2018;34(3):201-211.].

Epithelial Markers aSMA, Krt14, and Krt19 Unveil Elements of Murine Lacrimal Gland Morphogenesis and Maturation.

As an element of the lacrimal apparatus, the lacrimal gland (LG) produces the aqueous part of the tear film, which protects the eye surface. Therefore, a defective LG can lead to serious eyesight impairment. Up to now, little is known about LG morphogenesis and subsequent maturation. In this study, we delineated elements of the cellular and molecular events involved in LG formation by using three epithelial markers, namely aSMA, Krt14, and Krt19. While aSMA marked a restricted epithelial population of the terminal end buds (TEBs) in the forming LG, Krt14 was found in the whole embryonic LG epithelial basal cell layer. Interestingly, Krt19 specifically labeled the presumptive ductal domain and subsequently, the luminal cell layer. By combining these markers, the Fucci reporter mouse strain and genetic fate mapping of the Krt14+ population, we demonstrated that LG epithelium expansion is fuelled by a patterned cell proliferation, and to a lesser extent by epithelial reorganization and possible mesenchymal-to-epithelial transition. We pointed out that this epithelial reorganization, which is associated with apoptosis, regulated the lumen formation. Finally, we showed that the inhibition of Notch signaling prevented the ductal identity from setting, and led to a LG covered by ectopic TEBs. Taken together our results bring a deeper understanding on LG morphogenesis, epithelial domain identity, and organ expansion.

EXTENSIVE LICHENOID REACTION IN A PATIENT UNDER TREATMENT FOR CHRONIC MYELOID LEUKAEMIA

Adverse oral mucosa reactions resulting from imatinib and/or allopurinol use is very uncommon. Herein we report a new case of a 70-year-old female patient who was under imatinib and allopurinol treatment for 6 months against chronic myeloid leukemia, which developed extensive lichenoid eruptions in the oral mucosa, in addition to multiple cutaneous reactions in the skin, foot with the presence of pruritic violaceous nodules and diffuse whitish plaques. Microscopic exam of the cutaneous biopsy revealed hyperkeratosis, acanthosis and an intense chronic inflammatory infiltration in the underlying connective tissue that disrupted epithelial basal cell layer. The medical history together with clinic and microscopic features led to a final diagnosis of lichenoid reaction because of imatinib and/or allopurinol use. Allopurinol was interrupted and imatinib changed for dasatinib. Systemic and local corticosteroid therapy started and after forty-five days of follow-up, the lesions completely disappeared.

Using corneal confocal microscopy to track changes in the corneal layers of dry eye patients after autologous serum treatment

BackroundIn vivo corneal confocal microscopy allows the examination of each layer of the cornea in detail and the identification of pathological changes at the cellular level. The purpose of this study was to identify the possible effects of a three‐month treatment with autologous serum eye‐drops in different corneal layers of patients with severe dry eye disease using corneal confocal microscopy.MethodsTwenty‐six patients with dry eye disease were included in the study. Corneal fluorescein staining was performed. The corneas of the right eyes were examined using in vivo corneal confocal microscopy before and after a three‐month treatment with autologous serum drops. The densities of superficial and basal epithelial cells, Langerhans cells, the keratocytes and activated keratocytes, the density of endothelial cells and the status of the sub‐basal nerve plexus fibres were evaluated.ResultsA significant decrease in corneal fluorescein staining was found after the three‐month autologous serum treatment (p = 0.0006). The basal epithelial cell density decreased significantly (p = 0.001), while the density of superficial epithelial cells did not change significantly (p = 0.473) nor did the number of Langerhans cells or activated keratocytes (p = 0.223; p = 0.307, respectively). There were no differences in the other corneal cell layers or in the status of the nerve fibres.ConclusionsThe results demonstrate the ability of corneal confocal microscopy to evaluate an improvement in the basal epithelial cell layer of the cornea after autologous serum treatment in patients with dry eye disease. More studies with longer follow‐up periods are needed to elucidate the suitability of corneal confocal microscopy to follow the effect of autologous serum treatment on nerve fibres or other corneal layers in dry eye disease patients.

Orbital Involvement and Ocular Surface Changes in IgG4-Related Systemic Disease

To evaluate ophthalmic involvement of systemic immunoglobulin-G4-related disease (IgG4-RD) and describe the changes in both ocular surface parameters and corneal subbasal nerve plexus. Twenty eyes of 10 patients with systemic IgG4-RD and 20 eyes of 10 control subjects were enrolled in this study. Demographic information, medical history, serum IgG4 levels, orbital imaging, and follow-up data of subjects were recorded. Ocular surface tests were carried out in the order of tear break-up time (BUT), lissamine green (LG) staining, Schirmer I test with anesthesia, and ocular surface disease index (OSDI) questionnaire for all participants. Corneal subbasal nerves and basal epithelial cell layer were evaluated using in vivo confocal microscopy. Among the 10 patients with IgG4-RD, 11 eyes of 7 patients had orbital involvement. Among these 7 patients with IgG4-related ophthalmic disease, 4 presented with painless eyelid or periorbital swelling, 2 with diplopia and restricted ocular motility, and 1 with proptosis. Patients with IgG4-RD had higher OSDI (5.9 ± 6.6 vs. 1.7 ± 2.4, P < 0.001) and LG staining scores (0.7 ± 1.0 vs. 0.0 ± 0.0, P = 0.011) and lower BUT (5.6 ± 1.4 vs. 10.2 ± 1.0, P < 0.001) and Schirmer values (11.9 ± 10.3 vs. 18.3 ± 4.4, P = 0.021) as compared with those of control subjects. Total nerve density and nerve fiber length were found to be significantly lower in patients with IgG4-RD. The orbit is frequently involved during the course of IgG4-RD. These patients should be evaluated in terms of ocular surface disease and dry eye, which may be associated with lacrimal gland and/or orbital nerve involvement.

Utility of a novel triple marker (combination of thyroid transcription factor 1, Napsin A, and P40) in the subclassification of non–small cell lung carcinomas using fine-needle aspiration cases

Personalized treatment of lung cancer requires an accurate subclassification of non-small cell lung carcinoma (NSCLC) into adenocarcinoma (ADC), squamous cell carcinoma (SqCC), and other subtypes. In poorly differentiated tumors especially on small fine-needle aspirate specimens, the subclassification could be difficult in certain cases. Our previous study using resected tumor tissue has shown that the combination of commonly used individual markers (thyroid transcription factor 1 [TTF-1], P40, and Napsin A) into a novel triple marker has high sensitivity and specificity in subclassification of NSCLC and also the advantage of using minimal tumor tissue. In this study, we further evaluated the utility of this novel triple marker using fine-needle aspirate cases. We included primary NSCLC, consisting of 37 SqCCs (primary, 35; metastasis, 2) and 50 ADCs (primary, 29; metastasis, 21), 12 metastatic ADCs of nonpulmonary primary, and 10 small cell lung carcinomas. The immunohistochemical patterns were semiquantitatively scored. In lung SqCCs and ADCs, the sensitivity and specificity of the triple marker were 100% and 97.1% and 86.0% and 100%, respectively. The triple marker showed no immunoreactivity in 12 metastatic nonpulmonary ADCs. In 10 small cell lung carcinomas, TTF-1 had focal positivity in 40% of cases. The limitations of the triple marker include staining of alveolar macrophages (by TTF-1 and Napsin A), basal layer of bronchial epithelial cells (by P40), and nonspecific cytoplasmic staining of TTF-1. Our study not only supports our previous finding using resected tumor specimens but also provides evidence that the triple marker can be used for cytological material and preserving tumor tissue for molecular testing.

Elevated expression of ABCB5 in ocular surface squamous neoplasia.

ATP-binding cassette subfamily B member 5 (ABCB5) is a new member of the ATP-binding cassette superfamily and has been reported as a novel marker for limbal stem cell (LSC), which is essential for corneal homeostasis. ABCB5 expression has also been discovered in the subpopulation of several cancer cells containing the cancer stem cell (CSC). However, the pathogenetic relationship between LSC and CSC and ABCB5 in the ocular surface squamous neoplasm (OSSN) is still entirely unknown. To improve understanding of the role of ABCB5 in OSSN, we performed immunohistochemistry for ABCB5 in nine OSSN case series. While expression of ABCB5 is restricted to the basal epithelial cell layer in the normal limbus, elevated expressions of ABCB5 were clearly observed in all OSSN, and there was some breadth in the range of intensity of ABCB5 expression. Interestingly, the elevated expression patterns of ABCB5 in OSSN could be classified in three categories: perivascular, marginal and diffuse patterns. Our findings demonstrated for the first time that the expression of ABCB5 was upregulated in OSSN and that elevated expression of ABCB5 may be involved in the pathogenesis of OSSN.

Confocal Microscopic Analysis of a Rabbit Eye Model of High-Incidence Recurrent Herpes Stromal Keratitis.

Using CJLAT, a chimeric herpes simplex virus (HSV-1) that produces a high incidence of herpes stromal keratitis (HSK) in latently infected rabbits, and in vivo confocal microscopy (CM), we characterized the cellular events that precede the development of HSK. Thirty days after infection, in vivo CM was performed daily for 10 days and then weekly for up to 80 days after infection. We detected 3 types of subclinical corneal lesions before HSK was clinically apparent: (1) small epithelial erosions; (2) regenerating epithelium overlying small cell infiltrates within the basal epithelial cell layer; and (3) dendritic-like cells within the basal epithelial layer overlying stromal foci containing infiltrating cells. Sequential in vivo CM observations suggested that subclinical foci resolved over time but were larger and more abundant with CJLAT than with wild-type HSV-1 McKrae. Active HSK was observed only with CJLAT and was initially associated with a large epithelial lesion overlying stromal immune cell infiltrates. These results suggest that replication in the cornea of reactivated virus from the trigeminal ganglia produces epithelial lesions, which recruit immune cell infiltrates into the basal epithelial layer and anterior stroma. The virus is usually cleared rapidly eliminating viral antigens before the arrival of the immune cells, which disperse. However, if the virus is not cleared rapidly, or if an additional reactivation results in an additional round of virus at the same site before the immune cells disperse, then the immune cells are stimulated and may induce an immunopathological response leading to the development of HSK.

Transient laminin beta 1a Induction Defines the Wound Epidermis during Zebrafish Fin Regeneration.

The first critical stage in salamander or teleost appendage regeneration is creation of a specialized epidermis that instructs growth from underlying stump tissue. Here, we performed a forward genetic screen for mutations that impair this process in amputated zebrafish fins. Positional cloning and complementation assays identified a temperature-sensitive allele of the ECM component laminin beta 1a (lamb1a) that blocks fin regeneration. lamb1a, but not its paralog lamb1b, is sharply induced in a subset of epithelial cells after fin amputation, where it is required to establish and maintain a polarized basal epithelial cell layer. These events facilitate expression of the morphogenetic factors shha and lef1, basolateral positioning of phosphorylated Igf1r, patterning of new osteoblasts, and regeneration of bone. By contrast, lamb1a function is dispensable for juvenile body growth, homeostatic adult tissue maintenance, repair of split fins, or renewal of genetically ablated osteoblasts. fgf20a mutations or transgenic Fgf receptor inhibition disrupt lamb1a expression, linking a central growth factor to epithelial maturation during regeneration. Our findings reveal transient induction of lamb1a in epithelial cells as a key, growth factor-guided step in formation of a signaling-competent regeneration epidermis.

Cataract Surgery in Patients with Fuchs' Endothelial Corneal Dystrophy: When to Consider a Triple Procedure

To ascertain preoperative and intraoperative factors that predict the need for endothelial keratoplasty (EK) in patients with Fuchs' endothelial corneal dystrophy (FECD) undergoing cataract surgery.Prospective, observational cohort study.Eighty-nine patients (89 eyes) with FECD who require cataract surgery.One month before cataract surgery, we assessed best-corrected visual acuity, contrast sensitivity, straylight, keratometry, ultrasonic pachymetry, intraocular pressure, 7 corneal features of FECD and cataract density at slit-lamp examination, and corneal backscatter using in vivo confocal microscopy (IVCM; Confoscan 4, NIDEK Technologies, Padova, Italy). After surgery, measurements were repeated at 1, 2, and 12 months. We used stepwise binary logistic regression analysis to evaluate 30 preoperative and 5 intraoperative parameters for their ability to predict the postoperative need for EK. Receiver operating characteristic (ROC) curves of the predictive factors were used to identify their optimal cutoff points.Central corneal thickness (CCT) and backscatter at the basal epithelial cell layer (EV).After cataract surgery, 35 (39%) of 89 eyes underwent EK to restore vision. Of all preoperative and intraoperative parameters, only CCT and EV were identified as significant factors, predictive of the need for EK. The area under the ROC curve of EV was significantly higher than that of CCT (P = 0.003), whereas a combination of both factors in a linear discriminant function did not improve the predictive value (P = 0.66). As optimal cutoff points, we chose 1894 scatter units for EV and 630 μm for CCT. Both cutoff points correspond with a specificity of 94% and represent sensitivity of 63% for EV and 40% for CCT.Backscatter at the basal epithelial cell layer measured by IVCM predicts the need for EK after cataract surgery in patients with FECD. As an indicator for the corneal hydration state, the EV improves patient selection for combined cataract surgery and EK. In deciding whether to perform a triple procedure, CCT remains a less effective, but adequate, alternative. Regardless of the predictive factor used, a tailor-made approach is recommended accounting for individuals' expectations.