SOX17 enables immune evasion of early colorectal adenomas and cancers.

A hallmark of cancer is the avoidance of immune destruction. This process has been primarily investigated in locally advanced or metastatic cancer1-3; however, much less is known about how pre-malignant or early invasive tumours evade immune detection. Here, to understand this process in early colorectal cancers (CRCs), we investigated how naive colon cancer organoids that were engineered in vitro to harbour Apc-null, KrasG12D and Trp53-null (AKP) mutations adapted to the in vivo native colonic environment. Comprehensive transcriptomic and chromatin analyses revealed that the endoderm-specifying transcription factor SOX17 became strongly upregulated in vivo. Notably, whereas SOX17 loss did not affect AKP organoid propagation in vitro, its loss markedly reduced the ability of AKP tumours to persist in vivo. The small fraction of SOX17-null tumours that grew displayed notable interferon-γ (IFNγ)-producing effector-like CD8+ T cell infiltrates in contrast to the immune-suppressive microenvironment in wild-type counterparts. Mechanistically, in both endogenous Apc-null pre-malignant adenomas and transplanted organoid-derived AKP CRCs, SOX17 suppresses the ability of tumour cells to sense and respond to IFNγ, preventing anti-tumour T cell responses. Finally, SOX17 engages a fetal intestinal programme that drives differentiation away from LGR5+ tumour cells to produce immune-evasive LGR5- tumour cells with lower expression of major histocompatibility complex class I (MHC-I). We propose that SOX17 is a transcription factor that is engaged during the early steps of colon cancer to orchestrate an immune-evasive programme that permits CRC initiation and progression.

Lymphoglandular Complex-Like Colorectal Carcinoma-A Series of 20 Colorectal Cases, Including Newly Reported Features of Malignant Behavior.

Distinguishing colon carcinoma that is surrounded by well-circumscribed lymphoid tissue from adenomas involving lymphoglandular complexes can be difficult. We assessed a multi-institutional international cohort of 20 colorectal carcinomas with associated prominent lymphoid infiltrates, which we referred to as lymphoglandular complex-like carcinoma (LGCC). We collected clinical and endoscopic features, including lesion size, endoscopic appearance, location, procedure, follow-up, AJCC stage, and mismatch repair status. We recorded the presence of the following histologic features: haphazard gland distribution, gland angulation, gland fusion, solid nest formation, single-cell formation, stromal desmoplasia, presence of lymphovascular invasion and perineural invasion, presence of lamina propria, cytologic atypia as low- or high-grade, presence of goblet cells in the invasive component, and the presence of a surface lesion. Most cases (9 of 13) were described endoscopically as sessile polyps with an average size of 1.56 cm. Most cases (90%) were associated with a surface lesion, of which the majority were tubular adenomas, though a subset was associated with sessile serrated lesions with dysplasia (3 of 18). All cases of LGCC demonstrated haphazard gland distribution and either gland angulation, fusion, or solid nest formation. A portion of cases demonstrated single-cell infiltration (35%) and desmoplasia (50%), and rarely lymphovascular invasion was present (5%). A subset (10%) of cases invaded beyond the submucosa. Deficient mismatch repair was present in 22% (2 of 9) of cases for which it was performed. In cases of colectomy or completion colectomy, nodal metastasis was present in 38% (3 of 8). No cases demonstrated disease recurrence or disease-specific mortality. Overall, LGCC represents an enigmatic subset of carcinomas that is important to distinguish from adenomas involving lymphoglandular complexes due to its varying prognostic outcomes.

Chronic metabolic stress drives developmental programs and loss of tissue functions in non-transformed liver that mirror tumor states and stratify survival.

Under chronic stress, cells must balance competing demands between cellular survival and tissue function. In metabolic dysfunction-associated steatotic liver disease (MASLD, formerly NAFLD/NASH), hepatocytes cooperate with structural and immune cells to perform crucial metabolic, synthetic, and detoxification functions despite nutrient imbalances. While prior work has emphasized stress-induced drivers of cell death, the dynamic adaptations of surviving cells and their functional repercussions remain unclear. Namely, we do not know which pathways and programs define cellular responses, what regulatory factors mediate (mal)adaptations, and how this aberrant activity connects to tissue-scale dysfunction and long-term disease outcomes. Here, by applying longitudinal single-cell multi -omics to a mouse model of chronic metabolic stress and extending to human cohorts, we show that stress drives survival-linked tradeoffs and metabolic rewiring, manifesting as shifts towards development-associated states in non-transformed hepatocytes with accompanying decreases in their professional functionality. Diet-induced adaptations occur significantly prior to tumorigenesis but parallel tumorigenesis-induced phenotypes and predict worsened human cancer survival. Through the development of a multi -omic computational gene regulatory inference framework and human in vitro and mouse in vivo genetic perturbations, we validate transcriptional (RELB, SOX4) and metabolic (HMGCS2) mediators that co-regulate and couple the balance between developmental state and hepatocyte functional identity programming. Our work defines cellular features of liver adaptation to chronic stress as well as their links to long-term disease outcomes and cancer hallmarks, unifying diverse axes of cellular dysfunction around core causal mechanisms.

Beta-Hydroxybutyrate Augments Oxaliplatin-Induced Cytotoxicity by Altering Energy Metabolism in Colorectal Cancer Organoids.

Deregulation of cellular metabolism has recently emerged as a notable cancer characteristic. This reprogramming of key metabolic pathways supports tumor growth. Targeting cancer metabolism demonstrates the potential for managing colorectal cancer. Beta-hydroxybutyrate (BOHB) acts as an acetyl-CoA source for the tricarboxylic acid (TCA) cycle, possibly redirecting energy metabolic pathways towards the TCA cycle that could enhance sensitivity to oxaliplatin, through the generation of reactive oxygen species (ROS). This study explores the potential of BOHB to enhance oxaliplatin's cytotoxic effect by altering the energy metabolism in colorectal cancer. The study employed advanced in vitro organoid technology, which successfully emulates in vivo physiology. The combination treatment efficacy of BOHB and oxaliplatin was evaluated via cell viability assay. The levels of key proteins involved in energy metabolism, apoptotic pathways, DNA damage markers, and histone acetylation were analyzed via Western Blot. ROS levels were evaluated via flow cytometer. Non-toxic doses of BOHB with oxaliplatin significantly amplified cytotoxicity in colorectal cancer organoids. Treatment with BOHB and/or melatonin resulted in significantly decreased lactate dehydrogenase A and increased mitochondrial carrier protein 2 levels, indicating inhibited aerobic glycolysis and an increased oxidative phosphorylation rate. This metabolic shift induced apoptotic cell death mediated by oxaliplatin, owing to high levels of ROS. Melatonin counteracted this effect by protecting cancer cells from high oxidative stress conditions. BOHB may enhance the efficacy of chemotherapeutics with a similar mechanism of action to oxaliplatin in colorectal cancer treatment. These innovative combinations could improve treatment outcomes for colorectal cancer patients.

Randomised, placebo-controlled, phase 3 trial of the effect of the omega-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) on colorectal cancer recurrence and survival after surgery for resectable liver metastases: EPA for Metastasis Trial 2 (EMT2) study protocol.

INTRODUCTION: There remains an unmet need for safe and cost-effective adjunctive treatment of advanced colorectal cancer (CRC). The omega-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) is safe, well-tolerated and has anti-inflammatory as well as antineoplastic properties. A phase 2 randomised trial of preoperative EPA free fatty acid 2 g daily in patients undergoing surgery for CRC liver metastasis showed no difference in the primary endpoint (histological tumour proliferation index) compared with placebo. However, the trial demonstrated possible benefit for the prespecified exploratory endpoint of postoperative disease-free survival. Therefore, we tested the hypothesis that EPA treatment, started before liver resection surgery (and continued postoperatively), improves CRC outcomes in patients with CRC liver metastasis. METHODS AND ANALYSIS: The EPA for Metastasis Trial 2 trial is a randomised, double-blind, placebo-controlled, phase 3 trial of 4 g EPA ethyl ester (icosapent ethyl (IPE; Vascepa)) daily in patients undergoing liver resection surgery for CRC liver metastasis with curative intent. Trial treatment continues for a minimum of 2 years and maximum of 4 years, with 6 monthly assessments, including quality of life outcomes, as well as annual clinical record review after the trial intervention. The primary endpoint is CRC progression-free survival. Key secondary endpoints are overall survival, as well as the safety and tolerability of IPE. A minimum 388 participants are estimated to provide 247 CRC progression events during minimum 2-year follow-up, allowing detection of an HR of 0.7 in favour of IPE, with a power of 80% at the 5% (two sided) level of significance, assuming drop-out of 15%. ETHICS AND DISSEMINATION: Ethical and health research authority approval was obtained in January 2018. All data will be collected by 2025. Full trial results will be published in 2026. Secondary analyses of health economic data, biomarker studies and other translational work will be published subsequently. TRIAL REGISTRATION NUMBER: NCT03428477.

A NEW ARTIFICIAL INTELLIGENCE-BASED CLINICAL DECISION SUPPORT SYSTEM FOR DIAGNOSIS OF MAJOR PSYCHIATRIC DISEASES BASED ON VOICE ANALYSIS.

BACKGROUND: Speech features are essential components of psychiatric examinations, serving as important markers in the recognition and monitoring of mental illnesses. This study aims to develop a new clinical decision support system based on artificial intelligence, utilizing speech signals to distinguish between bipolar, depressive, anxiety and schizophrenia spectrum disorders. SUBJECTS AND METHODS: A total of 79 patients, who were admitted to the psychiatry clinic between 2020-2021, including 15 with schizophrenia spectrum disorders, 24 with anxiety disorders, 25 with depressive disorders, and 15 with bipolar affective disorder, alongside with 25 healthy individuals were included in the study. The speech signal dataset was created by recording participants' readings of two texts determined by the Russell emotion model. The number of speech samples was increased by using random sampling in speech signals. The sample audio signals were decomposed into time-frequency coefficients using Wavelet Packet Transform (WPT). Feature extraction was performed using each coefficient obtained from both Mel-Frequency Cepstral Coefficients (MFCC) and Gammatone Cepstral Coefficient (GTCC) methods. The disorder classification was carried out using k-Nearest Neighbor (kNN) and Support Vector Machine (SVM) classifiers. RESULTS: The success rate of the developed model in distinguishing the disorders was 96.943%. While the kNN model exhibited the highest performance in diagnosing bipolar disorder, it performed the least effectively in detecting depressive disorders. Whereas, the SVM model demonstrated close and high performance in detecting anxiety and psychosis, but its performance was low in identifying bipolar disorder. The findings support the utilization of speech analysis for distinguishing major psychiatric disorders. In this regard, the future development of artificial intelligence-based systems has the potential to enhance the psychiatric diagnosis process.

Does corneal tattooing affect the conjunctival microbiota?

PURPOSE: This study aimed to investigate the effects of commercial tattoo inks used in corneal tattooing on conjunctival microbiota. METHOD: This prospective case control study consisted of 125 participants divided in the following three groups: 35 patients with corneal tattoos, 40 patients with corneal leukoma, and 50 healthy subjects. Corneal tattooing was performed in all the cases in this study using a tattoo pen machine and commercial tattoo ink. A total of 500 cultures were taken from 250 eyes of 125 individuals on chocolate and sheep blood agar. Bacteriological samples were taken from the inferior eyelid conjunctiva using a sterile cotton swab. Without any contact elsewhere, the swabs were smeared on bedside chocolate agars and 5% sheep blood agar. RESULTS: In tattooed eyes, bacterial growth was detected in 42.9% of the chocolate and sheep blood agar samples. In other healthy eyes of patients with corneal tattoos, 54.5% bacterial growth on chocolate agar and 57.1% on sheep blood agar were detected. No statistical difference was detected in the conjunctival microbiota of chocolate and sheep blood agar (p = 0.254, p = 0.134, respectively) in the tattooed eyes compared to the other eye of the individual. No statistically significant difference was found in terms of bacterial growth in tattooed, leukoma, or healthy eyes on chocolate and sheep blood agar (p = 0.408, p = 0.349). The growth rate of Staphylococcus epidermidis decreased by 33.3% (from 12 to 8) on chocolate agar in 35 tattooed eyes, and it decreased by 28.5% (from 14 to 10) on sheep blood agar, while gram-negative bacteria Brevundimonas diminuta, Acinetobacter lwoffii, and Psychrobacter faecalis were detected in three patients. CONCLUSION: Corneal tattooing using commercial dye does not affect conjunctival microbiota. In the past 3 years, 120 patients have been tattooed with commercial tattoo ink in Istanbul Medeniyet University Göztepe Training and Research Hospital. No complications related to infection were found in the 3-year follow-up. The gram-negative bacteria detected in the healthy control group and tattooed eyes were bacteria found on normal skin or in the respiratory tract. Although some gram-negative bacteria do not cause infection, careful eye examination, follow-up, and culture are required in suspicious cases.

Bacterial contamination of multi-use tear drops, gels, and ointments.

PURPOSE: This study aimed to investigate the bacterial contamination of multi-use tear drops, gels, and ointments that patients use at home. METHOD: A total of 271 multi-use containers used by 168 patients were examined. Conjunctival culture samples were obtained from patients who used tear drops, gels, and ointments that were found to be contaminated. RESULTS: Bacterial contamination was detected in 33 (12.2 %) out of the 271 containers. The contamination rate was 7.9 % in tear drops, 11.7 % in gels, and 32 % in ointments. A statistically significant difference was found between the drops, gels, and ointment groups (P = 0.04). Bacterial contamination was detected in 25 (18.9 %) out of 132 collapsible tubes and 8 (5.8 %) out of 139 plastic bottles (P = 0.01). Important bacteria, including Pseudomonas stutzeri, Pseudomonas aeruginosa, Bacillus licheniformis, Paenibacillus pabuli, Proteus mirabilis, Pantoea agglomerans, Morganella morganii, Serratia marcescens, and Serratia liquefaciens, were detected. Mucorales spp. fungus was seen in a gel. Staphylococcus epidermidis, methicillin-resistant Staphylococcus aureus, and M. morganii were found in the conjunctival microbiota of three patients. CONCLUSION: The overall contamination rate of ocular lubricants was low (12.2%); however, a significant difference was found between the drops, gels, and ointments. The contamination rate was higher in gels and ointments than that in drops. The contamination rate was found to be increased in the collapsible tube. The use of ocular lubricants is safe; however, patients must be cautious when using multi-use tear drops, gels, and ointments to avoid contamination. Whenever possible, bottles should be preferred instead of collapsible tubes.

Evaluation of the effect of unilateral and bilateral inferior oblique myectomy on fundus torsion in primary and secondary inferior oblique overaction.

OBJECTIVE: This article evaluates the effects of unilateral and bilateral inferior oblique myectomy (IOM) on fundus torsion in primary and secondary inferior oblique overaction (IOOA). METHODS: This study analyzed 230 OCT images of 53 eyes of 32 patients who had undergone IOM by a single surgeon in the last two years. The disc-foveal angle (DFA) was calculated by digitally measuring the angle between the horizontal line passing through the geometric center of the optic disc and the curved line connecting the fovea to the geometric center of the optic disc. DFA was classified into intorsion, normal torsion, and extortion. The DFA was measured from the OCT images before the operation in the first week, first month, third month, and sixth month. RESULTS: When all the patients in our study were evaluated together, IOM statistically reduced the mean DFA in the third month (p=0.00). The DFA was higher in the secondary IOOA group than in the primary IOOA group (p=0.24). Bilateral IOM statistically significantly reduced DFA in the third month (p=0.00) and decreased the DFA difference between the two eyes in the third month (p=0.583). Unilateral IOM increased the DFA, rather than decreasing it, in the first week in operated eyes (p=0594) and increased the DFA difference between the two eyes after surgery (p=0.477). When we evaluated the localization of the macula as an intorsion, normal intorsion, or extortion, the extortion decreased from 36 to nine in the third month after bilateral IOM, and intorsion was seen in only two. Unilateral surgery did not significantly change fundus torsion in primary IOOA, and it caused intorsion in 3 of 6 (50%) operated eyes in secondary IOOA. CONCLUSION: Although unilateral IOM provides a clinical improvement in secondary IOOA, it increases the difference in DFA between both eyes and causes intorsion in 50% of patients. Masked IOOA was detected in 3 of 11 (27.3%) patients who underwent unilateral IOM. When deciding on unilateral surgery, the possibility of increased DFA difference between both eyes, intorsion in the operated eye, and masked IOOA in the other eye should be considered.

Dietary fat and lipid metabolism in the tumor microenvironment.

Metabolic reprogramming has been considered a core hallmark of cancer, in which excessive accumulation of lipids promote cancer initiation, progression and metastasis. Lipid metabolism often includes the digestion and absorption of dietary fat, and the ways in which cancer cells utilize lipids are often influenced by the complex interactions within the tumor microenvironment. Among multiple cancer risk factors, obesity has a positive association with multiple cancer types, while diets like calorie restriction and fasting improve health and delay cancer. Impact of these diets on tumorigenesis or cancer prevention are generally studied on cancer cells, despite heterogeneity of the tumor microenvironment. Cancer cells regularly interact with these heterogeneous microenvironmental components, including immune and stromal cells, to promote cancer progression and metastasis, and there is an intricate metabolic crosstalk between these compartments. Here, we focus on discussing fat metabolism and response to dietary fat in the tumor microenvironment, focusing on both immune and stromal components and shedding light on therapeutic strategies surrounding lipid metabolic and signaling pathways.

Ultrasensitive Detection of Circulating LINE-1 ORF1p as a Specific Multicancer Biomarker.

Improved biomarkers are needed for early cancer detection, risk stratification, treatment selection, and monitoring treatment response. Although proteins can be useful blood-based biomarkers, many have limited sensitivity or specificity for these applications. Long INterspersed Element-1 (LINE-1) open reading frame 1 protein (ORF1p) is a transposable element protein overexpressed in carcinomas and high-risk precursors during carcinogenesis with negligible expression in normal tissues, suggesting ORF1p could be a highly specific cancer biomarker. To explore ORF1p as a blood-based biomarker, we engineered ultrasensitive digital immunoassays that detect mid-attomolar (10-17 mol/L) ORF1p concentrations in plasma across multiple cancers with high specificity. Plasma ORF1p shows promise for early detection of ovarian cancer, improves diagnostic performance in a multianalyte panel, provides early therapeutic response monitoring in gastroesophageal cancers, and is prognostic for overall survival in gastroesophageal and colorectal cancers. Together, these observations nominate ORF1p as a multicancer biomarker with potential utility for disease detection and monitoring. SIGNIFICANCE: The LINE-1 ORF1p transposon protein is pervasively expressed in many cancers and is a highly specific biomarker of multiple common, lethal carcinomas and their high-risk precursors in tissue and blood. Ultrasensitive ORF1p assays from as little as 25 µL plasma are novel, rapid, cost-effective tools in cancer detection and monitoring. See related commentary by Doucet and Cristofari, p. 2502. This article is featured in Selected Articles from This Issue, p. 2489.

Ablative radiotherapy improves survival but does not cure autochthonous mouse models of prostate and colorectal cancer.

BACKGROUND: Genetically engineered mouse models (GEMMs) of cancer are powerful tools to study mechanisms of disease progression and therapy response, yet little is known about how these models respond to multimodality therapy used in patients. Radiation therapy (RT) is frequently used to treat localized cancers with curative intent, delay progression of oligometastases, and palliate symptoms of metastatic disease. METHODS: Here we report the development, testing, and validation of a platform to immobilize and target tumors in mice with stereotactic ablative RT (SART). Xenograft and autochthonous tumor models were treated with hypofractionated ablative doses of radiotherapy. RESULTS: We demonstrate that hypofractionated regimens used in clinical practice can be effectively delivered in mouse models. SART alters tumor stroma and the immune environment, improves survival in GEMMs of primary prostate and colorectal cancer, and synergizes with androgen deprivation in prostate cancer. Complete pathologic responses were achieved in xenograft models, but not in GEMMs. CONCLUSIONS: While SART is capable of fully ablating xenografts, it is unable to completely eradicate disease in GEMMs, arguing that resistance to potentially curative therapy can be modeled in GEMMs.

Mice can be used to model the types of cancer seen in people to investigate the effects of cancer therapies, such as radiation. Here, we apply radiation therapy treatments that are able to cure cancer in humans to mice that have cancer of the prostate or colorectum. We show that the mice do not experience many side effects and that the tumours reduce in size, but in some cases show progression after treatment. Our study demonstrates that mice can be used to better understand how human cancers respond to radiation treatment, which can lead to the development of improved treatments and treatment schedules.

A cross-cultural adaptation and validation of the Turkish version of American Orthopaedic Foot and Ankle Society Metatarsophalangeal-Interphalangeal Scale (AOFAS-MTP-IP) for the hallux.

OBJECTIVES: In this study, we aimed to translate and culturally adapt the American Orthopaedic Foot and Ankle Society (AOFAS) Hallux Metatarsophalangeal (MTP)-Interphalangeal (IP) scale, which is used for the clinical assessment of patients with hallux valgus (HV), into Turkish and to evaluate its validity and reliability. PATIENTS AND METHODS: Between February 2022 and October 2022, a total of 67 patients (18 males, 49 females; mean age: 51.5±15.9 years; range, 18 to 68 years) with HV deformity and able to communicate in Turkish were included. Following the translation of the AOFAS hallux MTP-IP scale into Turkish, its cultural appropriateness was confirmed. Intra-rater and inter-rater reliabilities were assessed by intraclass correlation coefficients (ICCs), using data collected by two orthopedists. Agreement among test-retest evaluations was conducted using the Bland-Altman analysis. The construct validity of the scale was determined by the Manchester-Oxford Foot Questionnaire (MOXFQ) and Short Form Health Survey (SF-36). Content validity was confirmed by the floor/ceiling effects. RESULTS: The Turkish AOFAS hallux MTP-IP had an excellent intra-rater reliability of 0.971. The intra-rater reliability of the pain, function, and alignment subscales ranged from 0.904 to 0.978. The inter-rater reliability was 0.913 for the total score, while ranging from 0.838 to 0.918 for the subscales. The total score of the AOFAS hallux MTP-IP had a high correlation with the physical domains of the MOXFQ and SF-36, while weaker correlations with mental domains were observed. No floor/ceiling effect was observed for the overall Turkish AOFAS hallux MTP-IP. CONCLUSION: The Turkish translated and culturally adapted AOFAS hallux MTP-IP scale is a valid and reliable measure, ensuring its use in assessing the clinical status of Turkish patients with HV deformity.

Immune microenvironment and lymph node yield in colorectal cancer.

BACKGROUND: Lymph node (LN) harvesting is associated with outcomes in colonic cancer. We sought to interrogate whether a distinctive immune milieu of the primary tumour is associated with LN yield. METHODS: A total of 926 treatment-naive patients with colorectal adenocarcinoma with more than 12 LNs (LN-high) were compared with patients with 12 or fewer LNs (LN-low). We performed immunohistochemistry and quantification on tissue microarrays for HLA class I/II proteins, beta-2-microglobulin (B2MG), CD8, CD163, LAG3, PD-L1, FoxP3, and BRAF V600E. RESULTS: The LN-high group was comprised of younger patients, longer resections, larger tumours, right-sided location, and tumours with deficient mismatch repair (dMMR). The tumour microenvironment showed higher CD8+ cells infiltration and B2MG expression on tumour cells in the LN-high group compared to the LN-low group. The estimated mean disease-specific survival was higher in the LN-high group than LN-low group. On multivariate analysis for prognosis, LN yield, CD8+ cells, extramural venous invasion, perineural invasion, and AJCC stage were independent prognostic factors. CONCLUSION: Our findings corroborate that higher LN yield is associated with a survival benefit. LN yield is associated with an immune high microenvironment, suggesting that tumour immune milieu influences the LN yield.

Aesthetic corneal tattooing/keratopigmentation using tattoo pen machine: choosing suitable method and color.

PURPOSE: This article aims to present the corneal tattooing method and how using a tattoo pen machine can improve aesthetic appearance in patients with corneal leukoma. METHODS: In this study, 42 patients were evaluated who had no visual potential and who had undergone colored corneal tattooing using an automatic tattoo pen machine for aesthetic purposes. The procedure was conducted according to the principles of the Declaration of Helsinki. The commercially available tattoo ink that has traditionally been used on human skin (brown, green, and black) for years was used for all the patients in this study, and 252 corneal photographs (with a Topcon slit lamp imaging device at 16 magnifications, i.e., 16 ×) taken within the last 2 years were evaluated retrospectively. Red, green, and blue (RGB) and hue, saturation, and lightness (HSL) values of the tattooed areas, such as pupils and iris, in corneal photographs were determined online using the Color Code Finder program. The RGB and HSL values of the pupil and iris were compared before surgery on the first day and first week, first month, third month, and twelfth month after surgery. RESULTS: In the first postoperative month, the mean pupil lightness (L) and iris L values were found to have increased by 10.7% and 5.7%, respectively. Between the first month and the first year, the L value of the mean pupil and that of the iris increased by 1.7% and 5.2%, respectively. The increase in the RGB value of the mean pupil in the first month was statistically significant (p = 0.02). The highest increase in RGB values of the iris was observed in the first week and first month (p = 0.113). This result shows that the majority of fading occurred in the first month. After the first month, the increase in the L value in the black-colored pupil was less than that in the brown- or green-colored iris. These results show that light colors fade faster and more. CONCLUSION: Esthetically, corneal leukoma causes severe psychological problems. Many patients are unable to use prosthetic contact lenses. Evisceration surgery has many complications, and limbal stem cells are used in evisceration surgery. Corneal tattooing using a tattoo pen machine is an easy, practical, and repeatable method used for aesthetic purposes. Successful results require the use of appropriate methods, ink, and ophthalmologist's experience. All patients in this study had a more aesthetic appearance than the preoperative white eye. Further studies are needed to develop a colored aesthetic tattooing method with a tattoo pen machine.