Early postoperative outcomes among patients with delayed surgeries after preoperative positive test for SARS-CoV-2: A case-control study from a single institution.

BACKGROUND: There are limited data on surgical complications for patients that have delayed surgery after severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. We aimed to analyze the surgical outcomes of patients submitted to surgery after recovery from SARS-CoV-2 infection. METHODS: Asymptomatic patients that had surgery delayed after preoperative reverse-transcription polymerase chain reaction (RT-PCR) for SARS-CoV-2 were matched in a 1:2 ratio for age, type of surgery and American Society of Anesthesiologists to patients with negative RT-PCR for SARS-CoV-2. RESULTS: About 1253 patients underwent surgical procedures and were subjected to screening for SARS-CoV-2. Forty-nine cases with a delayed surgery were included in the coronavirus disease (COVID) recovery (COVID-rec) group and were matched to 98 patients included in the COVID negative (COVID-neg) group. Overall, 22 (15%) patients had 30-days postoperative complications, but there was no statistically difference between groups -16.3% for COVID-rec and 14.3% for COVID-neg, respectively (odds ratio [OR] 1.17:95% confidence interval [CI] 0.45-3.0; p = .74). Moreover, we did not find difference regarding grades more than or equal to 3 complication rates - 8.2% for COVID-rec and 6.1% for COVID-neg (OR 1.36:95%CI 0.36-5.0; p = .64). There were no pulmonary complications or SARS-CoV-2 related infection and no deaths within the 30-days after surgery. CONCLUSIONS: Our study suggests that patients with delayed elective surgeries due to asymptomatic preoperative positive SARS-CoV-2 test are not at higher risk of postoperative complications.

LINE-1 hypomethylation and mutational status in cutaneous melanomas.

Epigenetic dysregulation is an important emerging hallmark of cutaneous melanoma development. The global loss of DNA methylation in gene-poor regions and transposable DNA elements of cancer cells contributes to increased genomic instability. Long interspersed element-1 (LINE-1) sequences are the most abundant repetitive sequence of the genome and can be evaluated as a surrogate marker of the global level of DNA methylation. In this work, LINE-1 methylation levels were evaluated in cutaneous melanomas and normal melanocyte primary cell cultures to investigate their possible association with both distinct clinicopathological characteristics and tumor mutational profile. A set of driver mutations frequently identified in cutaneous melanoma was assessed by sequencing (actionable mutations in BRAF, NRAS, and KIT genes, and mutations affecting the TER T promoter) or multiplex ligation-dependent probe amplification (MLPA) (CDKN2A deletions). Pyrosequencing was performed to investigate the methylation level of LINE-1 and CDKN2A promoter sequences. The qualitative analysis showed a trend toward an association between LINE-1 hypomethylation and CDKN2A inactivation (p=0.05). In a quantitative approach, primary tumors, mainly the thicker ones (>4 mm), exhibited a trend toward LINE-1 hypomethylation when compared with control melanocytes. To date, this is the first study reporting in cutaneous melanomas a possible link between the dysregulation of LINE-1 methylation and the presence of driver mutations.

DNA Methylation Levels of Melanoma Risk Genes Are Associated with Clinical Characteristics of Melanoma Patients.

In melanoma development, oncogenic process is mediated by genetic and epigenetic mutations, and few studies have so far explored the role of DNA methylation either as predisposition factor or biomarker. We tested patient samples for germline CDKN2A methylation status and found no evidence of inactivation by promoter hypermethylation. We have also investigated the association of clinical characteristics of samples with the DNA methylation pattern of twelve genes relevant for melanomagenesis. Five genes (BAP1, MGMT, MITF, PALB2, and POT1) presented statistical association between blood DNA methylation levels and either CDKN2A-mutation status, number of lesions, or Breslow thickness. In tumors, five genes (KIT, MGMT, MITF, TERT, and TNF) exhibited methylation levels significantly different between tumor groups including acral compared to nonacral melanomas and matched primary lesions and metastases. Our data pinpoint that the methylation level of eight melanoma-associated genes could potentially represent markers for this disease both in peripheral blood and in tumor samples.

LINE-1 hypermethylation in peripheral blood of cutaneous melanoma patients is associated with metastasis.

Aberrant DNA methylation pattern is a well-known epigenetic marker of cancer cells. Recently, aberrant methylation was also reported in the peripheral blood of cancer patients and it could potentially serve as a biomarker for cancer risk. We investigated the methylation pattern of LINE-1 and other repetitive DNA elements in peripheral blood of cutaneous melanoma patients in order to search for an association with clinical characteristics. The patient cohort was composed by 69 unrelated melanoma patients, 28 of whom were hereditary cases (with or without CDKN2A mutations) and 41 were isolated (sporadic) melanoma cases. Methylation of LINE-1 was evaluated by pyrosequencing, whereas additional repetitive DNA sequences were assessed using Illumina 450K methylation microarray. Melanoma patients exhibited a higher, albeit heterogeneous, LINE-1 methylation level compared with controls. Hereditary melanoma patients carrying CDKN2A mutations showed a hypermethylated pattern of both LINE-1 and repetitive DNA elements compared with other patients. In particular, the methylation level at one specific CpG of LINE-1 was found to be correlated with the occurrence of metastasis. Our data suggest that LINE-1 hypermethylation in peripheral blood of melanoma patients is a potential epigenetic biomarker for metastasis occurrence.

Gene network analyses point to the importance of human tissue kallikreins in melanoma progression.

BACKGROUND: A wide variety of high-throughput microarray platforms have been used to identify molecular targets associated with biological and clinical tumor phenotypes by comparing samples representing distinct pathological states. METHODS: The gene expression profiles of human cutaneous melanomas were determined by cDNA microarray analysis. Next, a robust analysis to determine functional classifications and make predictions based on data-oriented hypotheses was performed. Relevant networks that may be implicated in melanoma progression were also considered. RESULTS: In this study we aimed to analyze coordinated gene expression changes to find molecular pathways involved in melanoma progression. To achieve this goal, ontologically-linked modules with coordinated expression changes in melanoma samples were identified. With this approach, we detected several gene networks related to different modules that were induced or repressed during melanoma progression. Among them we observed high coordinated expression levels of genes involved in a) cell communication (KRT4, VWF and COMP); b) epidermal development (KLK7, LAMA3 and EVPL); and c) functionally related to kallikreins (EVPL, KLK6, KLK7, KLK8, SERPINB13, SERPING1 and SLPI). Our data also indicated that hKLK7 protein expression was significantly associated with good prognosis and survival. CONCLUSIONS: Our findings, derived from a different type of analysis of microarray data, highlight the importance of analyzing coordinated gene expression to find molecular pathways involved in melanoma progression.

Morphologic and clinical effects of subretinal injection of indocyanine green and infracyanine green in rabbits.

PURPOSE: The aim of this study was to determine the effects of subretinal injection of indocyanine green (ICG), infracyanine (IfCG), and balanced salt solution (BSS) in rabbits. METHODS: Ten (10) animals were subjected to a subretinal injection of 0.05% ICG (279 mOsm), 0.5% IfCG (276 mOsm), and BSS (300 mOsm) used as a control. Animals were examined at 6, 12, and 24 h and 14 days following the surgical procedure by indirect binocular ophthalmoscopy, fluorescein angiography (FA), and light and transmission electron microscopy. RESULTS: The subretinal injection of ICG caused damage to all retinal layers and retinal pigment epithelium (RPE) during the entire follow-up. Subretinal injection of IfCG resulted in abnormalities of the photoreceptor outer segments (POSs) during the entire follow-up; however, abnormalities of the photoreceptor inner segments (PISs) and outer nuclear layer (ONL) were observed only 24 h and 14 days after surgery; no RPE damage was observed. FA showed that window defects were more prominent in the subretinal ICG bleb position than the IfCG-related area. BSS caused only abnormalities of the POS layer and no RPE alterations. CONCLUSIONS: Subretinal injection of 0.05% ICG results in more significant retinal damage than 0.5% IfCG. In this model, iodine-free IfCG demonstrates a safer profile than a tenfold lower concentration of ICG, which contains iodine in its composition.

Sentinel lymph node biopsy in cutaneous melanoma: analysis of 240 consecutive cases.

BACKGROUND: The objective of this study was to evaluate practical rules for sentinel lymph node biopsy for melanoma and discuss the indications and outcomes of 240 patients. METHODS: A prospective, nonrandomized analysis was performed on 240 patients in a referral cancer center. The median patient age was 51 years, and the median Breslow thickness was 1.60 mm. Ulceration was found in 30.4 percent of the cases. The median follow-up was 27.81 months. The sentinel lymph node biopsy was performed in 240 patients with cutaneous melanoma thicker or equal to 1 mm. The operation was performed with preoperative lymphoscintigraphy and postoperative immunohistochemistry. A statistical analysis was performed comparing the need for a gamma probe in each location, the value of the experience, the need for immunohistochemistry, positivity compared with Breslow thickness, reasons for the success of the lymph node localization, and evolution. RESULTS: A total of 263 lymph node basins were identified (160 in the axilla, 86 in the inguinal region, and 17 in less common locations, including the popliteal, epitrochlear, and cervical regions). In every lymph node basin, the success of localization was directly related to use of the probe. The success rate for finding the sentinel lymph node increased year by year. Lymph node analysis disclosed positivity of 12.5 percent with hematoxylin and eosin staining and 17.5 percent with immunohistochemistry (excluding the sentinel lymph node not found disclosed 13.2 percent with hematoxylin and eosin and 18.5 percent with HMB45). Immunohistochemistry increased positivity by 40 percent. Positivity was directly related to Breslow thickness (p < 0.001). CONCLUSIONS: This study shows the importance of the gamma probe in all lymph node basins but mainly in the axilla and unusual basins, as well as the importance of experience and immunohistochemistry. As a new procedure, it was possible to recognize the pattern of recurrence in the follow-up.