Molecular identification, phylogenetic analysis and histopathological study of pathogenic free-living amoebae isolated from discus fish (Symphysodon aequifasciatus) in Iran: 2020-2022.

Free-living amoebae (FLA) are capable of inhabiting diverse reservoirs independently, without relying on a host organism, hence their designation as "free-living". The majority of amoebae that infect freshwater or marine fish are amphizoic, or free-living forms that may colonize fish under particular circumstances. Symphysodon aequifasciatus, commonly referred to as the discus, is widely recognized as a popular ornamental fish species. The primary objective of the present study was to determine the presence of pathogenic free-living amoebae (FLA) in samples of discus fish. Fish exhibiting clinical signs, sourced from various fish farms, were transferred to the ornamental fish clinic. The skin, gills, and intestinal mucosa of the fish were collected and subjected to culturing on plates containing a 1% non-nutrient agar medium. The detection of FLA was conducted through morphological, histopathological and molecular methods. The construction of the phylogenetic tree for Acanthamoeba genotypes was achieved using the maximum likelihood approach. The molecular sequence analysis revealed that all cultures that tested positive for FLA were T4 genotype of Acanthamoeba and Acanthamoeba sp. The examination of gill samples using histopathological methods demonstrated the presence of lamellar epithelial hyperplasia, significant fusion of secondary lamellae, and infiltration of inflammatory cells. A multitude of cysts, varying in shape from circular to elliptical, were observed within the gills. The occurrence of interlamellar vesicles and amoeboid organisms could be observed within the epithelial tissue of the gills. In the current study, presence of the Acanthamoeba T4 genotype on the skin and gills of discus fish exhibiting signs of illness in freshwater ornamental fish farms was identified. This observation suggests the potential of a transmission of amoebic infection from ornamental fish to humans, thereby highlighting the need for further investigation into this infection among ornamental fish maintained as pets, as well as individuals who interact with them and their environment.

The role of viruses in oral mucosal lesions.

The mucosa of the oral cavity is exposed to a large number of different microorganisms such as archaea, bacteria, fungi, parasites, and viruses. Among those, viruses cause specific infections, which can easily be transmitted from one person to another. The infectious route may not only include patients and their relatives but also the dental professional team. Thus, a wide knowledge regarding specific viral infections is crucial for the daily routine. Signs and symptoms of oral viral infections can be completely absent or develop into a pronounced clinical picture, so that early detection and information determine the further course of the infection and its influence on other inflammatory diseases, such as periodontitis, as well as the safety of family members and the social environment. As the clinical manifestation of viral infections may be highly variable leading to heterogenous mucosal lesions it is, in most cases, mandatory to differentiate them by specific microbiological tests in addition to clinical examination procedures. This article will give an overview of the role of viruses infecting the oral mucosa, and in addition, describe their clinical manifestation and management.

Histopathologic and Preneoplastic Changes in Tubal Ligation Materials.

Background and Objectives: To investigate histopathological changes and serous carcinoma precursors such as secretory cell outgrowths (SCOUTs) and p53 signature in the bilateral tubal ligation (BTL) materials used during cesarean section (S/C). Materials and Methods: In total, 138 patients underwent S/C and tubal sterilization (TS) between October 2020 and May 2021 at Konya City Hospital. Patients' data were obtained from the hospital's system. All data and findings were investigated and statistically evaluated. Results: The mean age was 34.62 years (22-44), the mean gravity was 4.89 (2-15) and the mean parity was 3.46 (1-10). In total, 5.79% SCOUT, 7.24% atypia and 9.42% p53 signatures were observed. Significant correlations were shown between the epithelial cell lineage and age between Ki-67, SCOUT, and gravity; between the Ki-67 results and gravity and parity; and between the p53 score and age. Conclusions: TS is a common, safe, and effective method worldwide. Today, BTL is increasing along with increasing S/C ratios. In addition to the reduced risk of ovarian cancers with ligation alone, precursor lesions such as hyperplasia, SCOUT, p53 signature, and STIL/Serous tubal intraepithelial carcinoma (STIC) are encountered in the ampulla materials obtained. Considering the low rates of re-anastomosis, tubal excision may be recommended instead of ligation in women of relatively higher gravity and age.

Glycyrrhizinic Acid as an Antiviral and Anticancer Agent in the Treatment of Human Papillomavirus.

Human papillomavirus (HPV), like any other virus, needs to penetrate the host cell and make use of its machinery to replicate. From there, HPV infection can be asymptomatic or lead to benign and premalignant lesions or even different types of cancer. HPV oncogenesis is due to the ability of the viral oncoproteins E6 and E7 to alter the control mechanisms for the growth and proliferation of host cell. Therefore, the use of agents with the ability to control these processes is essential in the search for effective treatments against HPV infections. Glycyrrhizinic acid (Gly), the active ingredient in liquorice, has been shown in numerous preclinical studies to have an antiviral and anticancer activity, reducing the expression of E6 and E7 and inducing apoptosis in cervical cancer cells. In addition, it also has antioxidant, anti-inflammatory, immunomodulatory or re-epithelializing properties that can be useful in HPV infections. This review includes the different antiviral and anticancer mechanisms described for Gly, as well as the clinical studies carried out that position it as a potential therapeutic strategy against HPV both through its topical application and by oral administration.

Multifocal epithelial hyperplasia confined to the interdental papilla of an adult Chinese man: a rare case report and literature review.

BACKGROUND: Multifocal epithelial hyperplasia (MEH), or focal epithelial hypertension (FEH), or Heck's disease, is an uncommon, benign oral mucosal disease associated with human papillomavirus infection. It is mostly observed in indigenous populations of the world, and has been rarely reported in China. However, previous research suggested there might be a greater prevalence of MEH in the Chinese population. While predominantly involves the lips, buccal mucosa and tongue, MEH was occasionally reported to affect the hard palate and gingiva as well. CASE PRESENTATION: This paper reports a case of extensive MEH lesions that confined to the interdental papilla of a Chinese male without detection of HPV, and summarizes the published gingiva-involved MEH reports from 1966 until present. The lesions were excised with an Er: YAG laser after scaling and root planning, no recurrence was observed after 6-month follow-up. CONCLUSIONS: The present report illustrates the need for clinicians to be aware of rare presentations of MEH to facilitate a prompt diagnosis and proper management. More reports are encouraged to determine a correct prevalence rate of MEH in China.

Anterior segment optical coherence tomography features of ocular surface squamous epithelial hyperplasia: A report of four cases.

Herein, we report the clinico-tomographic and histopathological features of four patients with biopsy-proven ocular surface squamous epithelial hyperplasia (OSSEH), a close mimicker of ocular surface squamous neoplasia (OSSN). The mean age at diagnosis was 58 years (median, 60 years; range, 35-77 years). All lesions were unilateral. Isolated corneal plaque was seen in 50% (n = 2) and nodular lesion at the nasal limbus in 50% (n = 2). Keratinization was seen in 75% (n = 3) of lesions and intrinsic vessels in 75% (n = 3). A clinical diagnosis of OSSN was made in all cases. Anterior segment optical coherence tomography (ASOCT) revealed mild epithelial hyperreflectivity in 100% (n = 4). The epithelium was normal in thickness in 75% (n = 3) and showed mild thickening in 25% (n = 1). Only 25% (n = 1) showed abrupt transition in epithelial thickness from the contiguous corneal epithelium. Histopathological examination revealed hyperplastic squamous epithelium and no cellular atypia in all cases. Stable ocular surface and no recurrences were noted at a mean follow-up of 17 months (median, 11 months; range, 2-43 months). Although OSSEH can mimic OSSN clinically, the presence of mild epithelial hyperreflectivity, lack of epithelial thickening, absence of abrupt transition from normal epithelium, and presence of subepithelial hyperreflectivity on ASOCT favor the diagnosis of OSSEH.

Photodynamic therapy in focal epithelial hyperplasia.

Focal epithelial hyperplasia (FEH) is an uncommon benign disorder affecting the oral mucosa. It is primarily associated with human papillomavirus (HPV) infection and presents as multiple white or pink soft papules or nodules. Typically, FEH is asymptomatic. Conventional treatment approaches for FEH include topical medication, surgical excision, CO2 laser ablation, cryotherapy, etc., but their efficacy varies. Photodynamic therapy (PDT) is a non-invasive and selective photochemotherapy method widely utilized in clinical practice. By employing specific light wavelengths to activate photosensitizers and induce the generation of reactive oxygen, PDT exerts cytotoxic effects. However, the application of PDT in treating FEH has not been previously documented. In this study, we present a case demonstrating the complete remission of FEH lesions using PDT, with no recurrence observed over a period of 9 months. This compelling outcome suggests that PDT may be a preferred treatment modality for FEH.

Mechanical removal of epithelial hyperplasia leads to successful treatment of irregular astigmatism.

BACKGROUND: Corneal epithelium remodeling in response to changes in the anterior corneal surface (keratoconus, corneal refractive surgery) is well-documented in the literature. However, several conditions may induce a different behavior of the epithelium, in which focal areas of epithelial thickening induce irregular astigmatism. This case report presents a highly unusual case of irregular astigmatism induced by an epithelial hyperplasia of unknown etiology, which was treated by the mechanical removal of only the epithelium. CASE PRESENTATION: A 29-year-old woman underwent implantable collamer lens implantation to correct myopia. The patient provided written informed consent. The procedure was uneventful in both eyes. Twenty months later, she complained of decreased visual acuity in the left eye (uncorrected distance visual acuity (UCDVA) was 20/30; corrected distance visual acuity was 20/20 with + 1.00 -2.25 × 170). Corneal topography revealed a nasal steepening in the left eye. Although the corneal thickness map was normal, epithelial thickness mapping revealed a localized nasal area of epithelial hyperplasia in the left eye that matched the area of steepest curvature. Slit lamp examination showed a total clear cornea with no signs of abnormality. The patient´s medical history was unremarkable and a case of epithelial hyperplasia of unknown etiology, without active inflammation, was considered. The decision was made to perform a mechanical removal of the corneal epithelium after application of diluted alcohol. One month after the procedure, the topography of the epithelized cornea showed a regular bow tie pattern and UCDVA improved to 20/20. No recurrence of the epithelial hyperplasia was detected after twenty-one months. CONCLUSIONS: Focal epithelial hyperplasia may induce irregular astigmatism. Epithelial thickness mapping is a very helpful technological tool to assess cases with irregular topography. De-epithelization as an isolated procedure may be useful for the successful management of these cases. Further research is required to understand the mechanism that triggers the spontaneous development of a focal epithelial hyperplasia.

Oral Human Papillomavirus Benign Lesions and HPV-Related Cancer in Healthy Children: A Systematic Review.

The present systematic review aimed to assess the prevalence of oral HPV-related lesions, categorized as benign (verruca vulgaris "VV", squamous cell papilloma "SP", condyloma acuminata "CA", and focal epithelial hyperplasia "FEH") and malignant (oral squamous cell carcinoma "OSCC"), in descending order of occurrence in pediatric subjects (≤18 years of age). The secondary objectives were to evaluate the frequency and types of oral lesions described in relation to HPV genotypes and the HPV vaccine type (if any). The study protocol, compliant with the PRISMA statement, was registered at PROSPERO (CRD42022352268). Data from 60 studies, of which quality was assessed using the ROBINS-I tool, were independently extracted and synthesized. Along with seven poorly described benign HPV-related oral lesions that could not be categorized, a total of 146 HPV-related oral lesions, namely 47.26% (n = 69) VV, SP, and CA, 51.37% (n = 75) FEH, and 1.37% (n = 2) OSSC, were diagnosed in 153 pediatric subjects (M:F ratio = 1:1.4) with a mean age of lesion onset of 8.46 years. The viral genotypes detected were HPV-13 (30.61%), -6 (20.41%), -11 (16.33%), HPV-2 (12.24%), -32 (10.20%), -57 (6.12%), and -16 (4.08%). No HPV vaccination was reported in any case. Further studies should be conducted to evaluate the prevalence of HPV-related benign and malignant lesions and the potential role of HPV and associated vaccination in oral carcinogenesis in pediatric subjects.

Refractive Effect of Epithelial Remodelling in Myopia after Transepithelial Photorefractive Keratectomy.

(1) Introduction: We analysed epithelial changes after the treatment of moderate myopia with transepithelial photorefractive keratectomy. (2) Materials and Methods: We used optical coherence tomography data and analysed changes in the stroma and epithelium after ablation. We aimed to ascertain how much epithelium hyperplasia occurred after TransPRK; for this, we used data from 50 eyes treated with TransPRK with the AMARIS 1050 Hz, with a minimum follow-up of 4 months. (3) Results: The measured epithelial changes corresponded to a less than 0.1 ± 0.2D of spherical effect, less than 0.2 ± 0.2D of astigmatic effect, and less than 0.5 ± 0.2D of comatic effect. (4) Conclusions: The changes in epithelial thickness after aberration-neutral transepithelial photorefractive keratectomy for moderate myopia were very small, indicating a low level of epithelial hyperplasia without resembling a regression-inducing lentoid.

Hypopigmented penile fixed drug eruption.

Fixed drug eruptions (FDE) are typically associated with residual hyperpigmentation or non-pigmenting lesions. There is no distinctive histopathological feature; though, drug provocation tests (DPT) can be confirmatory within 7 days. We describe a patient with penile FDE associated with residual hypopigmentation, a prolonged refractory period to DPT and recurrent meatal stenosis.

Anemoside B4 prevents chronic obstructive pulmonary disease through alleviating cigarette smoke-induced inflammatory response and airway epithelial hyperplasia.

BACKGROUND: Cigarette smoke (CS) is one of the major risk factors for chronic obstructive pulmonary disease (COPD) and increases the risk of lung cancer (LC). Anemoside B4 (B4) is the main bioactive ingredient in Pulsatilla chinensis (P. chinensis), a traditional medicinal herb for various diseases. It has a wide range of anti-inflammatory, anti-oxidation and anti-cancer activities. However, in recent years, there is no relevant literature report on the therapeutic effect of B4 on COPD, and the anti-inflammatory and inhibitory effects of anemoside B4 on basal cell hyperplasia in CS-induced COPD have not been clearly established. PURPOSE: In the present study, we investigated whether anemoside B4 could alleviate CS or cigarette smoke extract (CSE) induced inflammation of COPD and further prevent basal cell hyperplasia, hoping to find its possible mechanism. METHODS: In this study, a COPD mouse model was established in C57BL mice by CS exposure 3 months. Bronchial pathology and basal cell hyperplasia were observed by HE staining and immunostaining. The contents of glutathione peroxidase catalase (GSH-PX), malondialdehyde (MDA) and superoxide dismutase (MPO) were determined by GSH-PX, MDA and SOD assay kits, respectively. 16HBE cells were cultured with 5% CSE with or without treatment with B4 (1, 10, 100 µM) or DEX (20 µM) in vitro. Cell viability was assessed by a cell counting kit 8 (CCK-8). Reactive oxygen species (ROS) generation was tested by DCFH-DA. Moreover, anti-inflammatory mechanism of anemoside B4 was further determined by pro-inflammatory cytokines production using RT-PCR. Protein expression levels of MAPK/AP-1/TGF-ß signaling pathway were measured by western blot. RESULTS: Anemoside B4 improved the lung function of mice, relieved lung inflammation and reduced the MDA, MPO and GSH-Px in the plasma. At the same time, B4 repressed the oxidative stress response and played a role in balancing the levels of protease and anti-protease. During the process of bronchial basal cell hyperplasia, B4 alleviated the degree of cell hyperplasia, and prevented further deterioration of hyperplasia through increased P53 and inhibited FHIT protein. In addition, B4 reduced ROS levels in human bronchial epithelial cells stimulated by CSE in vitro study. Meanwhile, B4 treatment also significantly attenuated increased IL-1ß, TGF-ß, IL-8 and TNF-α from CSE treated human bronchial epithelial cells. The expression of p-P38, AP-1(c-fos, and c-Jun), TGF-ß proteins in MAPK/AP-1/TGF-ß signaling pathway were decreased and the signal cascade reaction was blocked. CONCLUSION: Anemoside B4 protects against CS-induced COPD. These findings indicated that B4 may have therapeutic potential for the prevention and treatment of COPD.

Useful cytological confirmation of HPV 13 in lesional mucosa enhances diagnosis of focal epithelial hyperplasia.

An 11-year-old female presented with multiple oral lesions for several months. Histopathological findings suggested focal epithelial hyperplasia (FEH), also known as Heck disease. FEH is strongly associated with Human papillomavirus (HPV), especially genotypes 13 and 32. An oral swab of a mucosal lesion was subsequently obtained for cytology, immunohistochemistry and in situ hybridization. In addition, in situ hybridization and immunohistochemistry were also performed retrospectively on the biopsy specimen for correlation. The cytology specimen showed squamous cells with enlarged, slightly atypical nuclei and rare perinuclear halos. The histology findings included papillomatosis with acanthosis, mild nuclear atypia and focal perinuclear halos. The immunohistochemistry for the consensus HPV L1 capsid protein was found in both the cytology and biopsy specimens indicating that the lesion was HPV-related. High viral copy numbers of HPV 13 were detected by in situ hybridization in both the cytology and histology specimens. Although histologic features of FEH have been well characterized in the literature, to our knowledge, this is the first case to describe in FEH with adjunct immunohistochemistry and in situ hybridization results. Furthermore, these findings assisted in our diagnosis since the patient's clinical presentation was a diagnostic challenge with smooth dome-shaped papules instead of the typically described flat-topped verrucous lesions seen in FEH. In summary, our case reveals that there is a high concordance between the HPV 13 detection in the cytology and histology of FEH, and that performing cytology in addition to histology can be used to optimize diagnostic evaluation towards appropriate patient care.

Diode Laser Excision of Focal Epithelial Hyperplasia (Heck's Disease): A Case Report.

Introduction: Focal epithelial hyperplasia (FEH), also known as Heck's disease, is a human papilloma virus induced proliferation of epithelial cells, which causes asymptomatic lesions in the oral mucosa. Heck's disease is more prevalent among children or in a family. Despite the self-limitation of lesions, they may progress or cause esthetical and functional discomforts. Hence, early diagnosis is necessary. Case Presentation: A middle-aged man with a peripheral multiple soft, elevated, sessile, smooth, lobulated surface and nodular lesions in his right buccal mucosa was referred. The dimensions of the lesions varied from 5 to 25 mm. Through clinical and histopathological examinations, Heck's disease was diagnosed. The diode laser (wavelengths of 980 nm), 1-W power using continuous waves) was applied serially (in 2 sessions with a 1-week interval) for surgically removing the lesions. The patient was followed up for 6 months with no lesion recurrence, and gradual healing was observed. Conclusion: The laser is an effective treatment with no scar remaining after the procedure and brings satisfaction to patients.

Hiperplasia epitelial focal: relato de caso clínico / Focal epithelial hyperplasia: case report

Introdução: a hiperplasia epitelial focal (HEF) ou doença de Heck, caracteriza-se por ser uma lesão benigna rara da mucosa bucal. Geralmente, está associada a proliferação do epitélio pavimentoso atribuída principalmente ao papiloma vírus humano (HPV) dos subtipos 13 e 32. Objetivo: relatar o caso de um paciente, com suspeita diagnóstica de HEF, descrever suas manifestações clínicas bucais e histopatológicas. Metodologia: homem, 38 anos, melanoderma, compareceu no Serviço Universitário com queixa de ardência em mucosa bucal. Ao exame clínico, apresentava múltiplas lesões papulares, firmes à palpação, de coloração rósea, com pápulas aglomeradas ou isoladas, localizadas em rebordo alveolares, gengiva inserida e livre da arcada superior e inferior, mucosas jugais e labiais, compatíveis com lesões causadas por HPV. Realizou-se biópsia incisional das lesões bucais, o laudo histopatológico descreveu o fragmento de mucosa bucal que estava revestida por epitélio pavimentoso estratificado paraceratinizado com acantose proeminente, projeções focais confluentes para o tecido conjuntivo, confirmando a suspeita diagnóstica clínica. Conclusão: a HEF é uma patologia rara, benigna da mucosa, indolor e que pode se resolver espontaneamente. Portanto, é necessário o diagnóstico adequado através da biopsia e laudo histopatológico.

Introduction: focal epithelial hyperplasia (FEH) or Heck's disease is characterized as a rare benign lesion of the oral mucosa. It is usually associated with proliferation of the squamous epithelium, mainly attributed to human papillomavirus (HPV) subtypes 13 and 32. Objective: to report the case of a patient with a diagnostic suspicion of HEF, to describe its oral clinical and histopathological manifestations. Methodology: hale, 38 years old, black, attended the University Service with a complaint of burning in the oral mucosa. On clinical examination, she presented multiple papular lesions, firm to palpation, pink in color, with agglomerated or isolated papules, located in the alveolar ridge, inserted gingiva free from the upper and lower arch, jugal and labial mucosa, compatible with lesions caused by HPV. An incisional biopsy of the oral lesions was performed, the histopathological report described the fragment of oral mucosa that was lined with parakeratinized stratified squamous epithelium with prominent acanthosis, focal projections confluent to the connective tissue, confirming the clinical diagnostic suspicion. Conclusion: FEH is a rare, benign mucosal pathology, painless and can resolve spontaneously. Therefore, proper diagnosis through biopsy and histopathological report is necessary.

Amniotic Membrane Grafting for Ocular Surface Inflammation Following Topical Interferon Alpha 2b Therapy.

We describe a rare case of a 58-year-old female with ocular surface squamous neoplasia (OSSN) in her left eye. She was treated for 12 months with topical interferon alpha-2b (IFNα-2b) eye drops and OSSN resolved completely. She presented with a whitish elevated lesion involving the cornea, limbus, and conjunctival surface after discontinuation of topical IFNα-2b. Excision biopsy along with amniotic membrane grafting was done to stabilize the ocular surface. Histopathological evaluation of the excised tissue revealed ocular surface inflammation with no evidence of malignancy.

An update on Heck's disease-a systematic review.

BACKGROUND: Previous research has suggested an ethnic association of Heck's disease with a prominent genetic and familial inheritance pattern, but no systematic review has been reported, which has collected all the evidence in one paper. The aim was estimation of the updated age estimates and gender predilection of this disease and also questioning its proposed link to ethnic and geographical factors. METHODS: Heck's disease from 1966 until present are tabulated, including various descriptive characteristics. After removal of duplicates and adhering to all the inclusion criteria, we shortlisted 95 case reports. The quality assessment of all included studies has been done following STROBE (STrengthening the Reporting of OBservational studies in Epidemiology) guidelines. RESULTS: We found an age range of 3-92 years (mean: 23.1 years) with a male to female ratio of 3:4. Geographical distribution revealed one of the main findings of this study, which was an increased incidence of Heck's disease in the European region. CONCLUSIONS: As already observed and established, there is a much greater prevalence of this disease in the indigenous populations of the world and more research should be encouraged to understand the correct transmission and pattern of spread of this disease.

FAK Shutdown: Consequences on Epithelial Morphogenesis and Biomarker Expression Involving an Innovative Biomaterial for Tissue Regeneration.

By employing an innovative biohybrid membrane, the present study aimed at elucidating the mechanistic role of the focal adhesion kinase (FAK) in epithelial morphogenesis in vitro over 4, 7, and 10 days. The consequences of siRNA-mediated FAK knockdown on epithelial morphogenesis were monitored by quantifying cell layers and detecting the expression of biomarkers of epithelial differentiation and homeostasis. Histologic examination of FAK-depleted samples showed a significant increase in cell layers resembling epithelial hyperplasia. Semiquantitative fluorescence imaging (SQFI) revealed tissue homeostatic disturbances by significantly increased involucrin expression over time, persistence of yes-associated protein (YAP) and an increase of keratin (K) 1 at day 4. The dysbalanced involucrin pattern was underscored by ROCK-IISer1366 activity at day 7 and 10. SQFI data were confirmed by quantitative PCR and Western blot analysis, thereby corroborating the FAK shutdown-related expression changes. The artificial FAK shutdown was also associated with a significantly higher expression of filaggrin at day 10, sustained keratinocyte proliferation, and the dysregulated expression of K19 and vimentin. These siRNA-induced consequences indicate the mechanistic role of FAK in epithelial morphogenesis by simultaneously considering prospective biomaterial-based epithelial regenerative approaches.