5-Methylcytosine immunohistochemistry for predicting cutaneous melanoma prognosis.

There is a correlation between DNA methylation and the diseased stage and poor survival. 5-methylcytosine (5-mC) is one of the epigenetic modifications of bases that researchers focus on. Staining with 5-mC immunohistochemistry was used to examine pathological samples taken from individuals diagnosed with cutaneous melanoma. Between Breslow levels 2 and 4, there was a significant difference in the H-score of 5-mC expression (p = 0.046). A significant reduction in 5-mC expression H-scores was seen in patients who were diagnosed with ulcers (p = 0.039). It was shown that patients with low 5-mC had a significantly worse overall survival rate (p = 0.027).

CpG methylation changes in Alu repetitive sequences in normal aging due to diastolic hypertension in human dermal fibroblasts from the facial area.

Aging fibroblasts, an important factor contributing to skin aging, are affected by numerous mechanisms, including alterations in DNA methylation and age-related diseases. The current study aimed to investigate the role of Alu methylation in aging fibroblasts and hypertension. The Alu methylation levels in dermal fibroblasts obtained from patients of different ages and blood pressure status were analyzed using the combined bisulfite restriction analysis technique. An inverse correlation was observed between Alu methylation in dermal fibroblasts and patient age. Dermal fibroblasts from the high-normal diastolic blood pressure group had higher Alu methylation levels compared with those from the normal group. The findings of the present study suggest that Alu methylation alterations can be observed with chronological aging and hypertension, and are a potential aging marker or therapeutic target.

Evaluation of P53 immunostaining in patients with cutaneous melanoma.

P53 is a tumor suppressor gene that is mutated in numerous types of cancer. The aim of the present study was to determine the frequency of this mutation in cutaneous melanomas and to conduct clinicopathological characteristics and clinical outcome association analyses with the P53 mutation. P53 immunohistochemical staining was used as a surrogate marker for P53 mutation analysis to assess P53 status. In the present study, 50 pathological samples of cutaneous melanoma from 2012 to 2018 at Chulalongkorn University (Bangkok, Thailand), were subjected to anti-P53 immunohistochemistry, followed by an examination of the association between P53 statuses and clinical and pathological characteristics, along with clinical outcomes. A positive staining for anti-P53 antibody was detected in 30% of patients (15/50) with cutaneous melanomas. Positivity was significantly associated with female sex, nodular histological subtype and Breslow level 4. Cox regression analysis revealed that an age >65.5 years and Breslow grade 4 disease were associated with mortality. The Kaplan-Meier curve revealed a shorter duration of recurrence time in the P53 mutation than P53 wild type. In the present study, P53 mutations in specific cases of cutaneous melanoma were identified. Notably, patients who were older and/or had a Breslow score of 4 exhibited an increased risk of mortality. These findings suggested the potential involvement of P53 mutations in cutaneous melanoma, highlighting the necessity for further investigations to improve understanding of their roles.

Weight to volume conversion: an easy and practical breast volume estimation.

Background: Accurate breast volume estimation is essential for symmetrical breast reconstruction. Easy conversion of the weight of the resected breast tissue to volume could result in precise volume measurements. This study aimed to introduce the use of a mathematical constant (k) to estimate the breast volume from the weight. Methods: Eighty-nine female patients with breast cancer who underwent surgery at King Chulalongkorn Memorial Hospital between September 2010 and February 2011 were enrolled in this prospective study. The mammographic density of each patient was classified according to the breast imaging reporting and data system (BI-RADS) into groups a, b, c, and d. The breast density number and mathematical constant (k) were calculated, and the data matched. This technique was validated by comparing the measured and calculated volumes. Results: Sixty-six, 22, and 1 patients underwent total mastectomies (TMs), skin-sparing mastectomies (SSMs), and nipple-sparing mastectomies (NSMs), respectively. The breast densities were 1.0629, 1.1545, and 1.2233 g/mL, and the constant number (k) was 0.9409, 0.8662, and 0.8175 for BI-RADS a, combined BI-RADS b and c, and BI-RADS d, respectively. The validation process showed no significant differences between the measured and calculated volumes [95% confidence interval (95% CI)]. The correlation coefficient (r) was 0.984. Conclusions: Accurate breast volume estimation is a key factor in achieving symmetry in breast reconstruction. Combining existing data, including the weight of the resected breast tissue and mammographic density findings, an easy and accurate method to calculate the resected breast volume was introduced.

BRAF V600E Immunohistochemistry Predicts Prognosis of Patients with Cutaneous Melanoma in Thai population.

The BRAF V600E mutation in the Thai population has been identified in a considerable percentage of people with cutaneous melanoma. The objectives of this study were to determine the prevalence of this mutation in cutaneous melanomas, conduct a clinicopathological association analysis with the BRAF V600E mutation, and develop a treatment strategy for patients with this mutation that would take advantage of the medications currently available to treat them. Methods: Anti-BRAF V600E (clone VE1) immunohistochemistry was performed on 50 pathological samples of cutaneous melanoma after excluding the samples with a low amount of pathologic tissue, a lack of clinical data' and poor follow-up. BRAF V600E expression DNA sequencing was performed to confirm the results of several cases. Results: Anti-BRAF V600E antibody positivity was noted in 56% (28/50) of cutaneous melanoma cases. DNA sequencing results were consistent with immunohistochemistry results. In cutaneous melanoma, the BRAF V600E mutation was significantly associated with adverse prognosis of patients, including reduced overall survival and disease-free survival. Conclusions: An increased prevalence of the BRAF V600E mutation was determined in a collection of cutaneous melanomas in the Thai population, implying that BRAF-targeted therapy may be a promising strategy for patients with BRAF-mutated cutaneous melanoma. This study revealed an association between the clinicopathological aspects of cutaneous melanoma and overall survival, disease-free survival, and overall mortality. A treatment with anti-BRAF-targeted therapy, which incorporates the already available medications' is being researched and developed.

Nutritional Support with Omega-3 Fatty Acids in Burn Patients: A Systematic Review with Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: In burn patients, the profound effect of nutritional support on improved wound healing and a reduced rate of hospitalization and mortality has been documented. Fish oil as a primary source of omega-3 fatty acids in nutritional support may attenuate the inflammatory response and enhance immune function; however, unclear effects on the improvement of clinical outcomes in burn patients remain. METHODS: The systematic literature review was conducted by searching the electronic databases: Cochrane Library, PubMed, ScienceDirect, and Scopus to assess the randomized controlled trials of nutritional support with omega-3 fatty acids compared to control diets in patients that presented with burns from any causes. RESULTS: Seven trials were included in this meta-analysis. We found no significant differences in length of stay (LOS) (p = 0.59), mortality (p = 0.86), ventilation days (p = 0.16), gastrointestinal complications-e.g., constipation and diarrhea (p = 0.73)-or infectious complications-e.g., pneumonia and sepsis (p = 0.22)-between the omega-3-fatty-acid-receiving group and the control/other diets group. CONCLUSIONS: We did not find a benefit of omega-3 support in reducing the various complications, mortality and LOS in burn patients. Further studies are necessary to find the effect of nutritional support with omega-3 fatty acids over low-fat diets in this population.

3D PED BURN app: A precise and easy-to-use pediatric 3D burn surface area calculation tool.

Background and aims: Calculating the precise total burn surface area is crucial when treating burn patients, particularly children. The Lund and Browder chart and Rule of Nines, 2-dimensional diagrams that are widely used, are subject to high interrater variance, and they can severely overestimate the burn area. Previously, the adult 3-dimensional burn area calculation mobile application was developed. Aiming to improve accuracy, a 3-dimensional pediatric burn surface area calculation mobile application ("3D PED BURN") was developed to overcome the limitations of the conventional methods. Method: Fifteen 3-dimensional pediatric burn surface area models based on detailed anthropometric measurements collected from 85 patients were developed and categorized into four age groups: <1 year; 1-4 years; 5-9 years, and 10-15 years. According to their weight and height, the models in each group were fractionated into large, medium, and small body sizes. Result: A precise and easy-to-use application was developed based on these data. This application is a promising and more accurate calculation tool for burn surface area in pediatric patients. Its low inter-rater variance makes it reliable for use by various healthcare personnel. Conclusion: The 3D PED BURN app is a pediatric 3D burn surface area calculation tool that is both accurate and simple to use.

B1 repetitive sequence methylation enhances wound healing of second-degree burns in rats.

The accumulation of DNA damage in burn wounds delays wound healing. DNA methylation by short interspersed nuclear element (SINE) small interfering (si)RNA prevents DNA damage and promotes cell proliferation. Therefore, SINE siRNA may be able to promote burn wound healing. Here, a SINE B1 siRNA was used to treat burn wounds in rats. Second-degree burn wounds were introduced on the backs of rats. The rats were then divided into three groups: a B1 siRNA-treated, saline-treated control, and saline + calcium phosphate-nanoparticle-treated control group (n=15/group). The wounds were imaged on days 0, 7, 14, 21 and 28 post-injury. The tissue sections were processed for methylation, histological and immunohistochemical examination, and scored based on the overall expression of histone H2AX phosphorylated on serine 139 (γH2AX) and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Burn wound closure improved in the B1 siRNA-treated group compared with that in the control group, especially from days 14-28 post-injury (P<0.001). The overall pathological score and degree of B1 methylation in the B1 siRNA-treated group improved significantly at days 14-28 post-injury, with the maximum improvement observed on day 14 (P<0.01) compared with the NSS and Ca-P nanoparticle groups. Immunohistochemical staining revealed lower expression of γH2AX and 8-OHdG in the B1 siRNA-treated group than in the control groups at days 14-28 post-injury; the maximum improvement was observed on days 14 and 21. These data imply that administering SINE siRNA is a promising therapeutic option for managing second-degree burns.

An anthropometric study of the external ear in the Thai population.

Auricular dimensions are important data for the treatment of auricular deformities, either congenital or acquired. Many previous studies have shown that the auricular dimension varies among ethnic groups; however, there are currently no available data for the Thai population. The objective of this investigation was to provide normative values regarding the adult ear dimensions in Thailand. A total of 200 Thai volunteers between the ages of 18 and 70 years (80 men, 120 women) were enrolled in this study. We measured six dimensions, including the auricular, lobular, and conchal length and width and the auricular projection at the superaurale and tragal levels. Moreover, we calculated the auricular, lobular, and conchal indices. The selected dimensions of the external ears were shown as means ± standard deviations. These parameters were then compared between the right and left sides, men and women and participants aged ≥50 and <50 years. We found that men have a significantly larger and more projected ear than women do. The length of the entire auricles and the length of the earlobe were also significantly longer in the population ≥50 years than those <50 years. This study provides normative data of the external ear dimension and projection in the adult Thai population. These data can be used to reconstruct the ear and face more appropriately for Thai individuals.

Alu repetitive sequence CpG methylation changes in burn scars.

Alu elements are retrotransposons related to epigenetic modifications. To date, the role of epigenetics in hypertrophic scars from burn remains unknown. Here, our aim was to examine the pathophysiology of hypertrophic scars from an epigenetic perspective. For that, we performed a cross-sectional analytical study using tissue and blood samples from burned and healthy patients (n = 23 each) to detect Alu methylation levels and patterns. The results of the combined bisulfite restriction analysis technique were categorized into four groups based on the methylation status at the CpG dinucleotides from the 5' to the 3' ends of the Alu sequence: hypermethylated (mCmC), hypomethylated (uCuC), and partially methylated (uCmC and mCuC). Alu methylation levels were significantly lower in hypertrophic scar tissues than in normal skin (29.37 ± 2.49% vs. 35.56 ± 3.18%, p = 0.0002). In contrast, the levels were significantly higher in white blood cells from blood samples of burned patients than in those of control blood samples (26.92 ± 4.04% vs. 24.58 ± 3.34%, p = 0.0278). Alu total methylation (mC) and the uCmC pattern were significantly lower, whereas uCuC was significantly higher, in hypertrophic scar tissues than in normal skin (p < 0.0001). Receiver operating characteristic analysis indicated that the uCmC and uCuC patterns are useful as hypertrophic scar DNA methylation markers after burn, with 91.30% sensitivity and 96.23% specificity and 100% sensitivity and 94.23% specificity, respectively. Our findings suggest that epigenetic modifications play a major role in hypertrophic scar pathogenesis, and may be the starting point for developing a novel technique for burn scar treatment.

The Effect of Cold Water on Pain Evaluation During Negative Pressure Wound Therapy Dressing Changes: A Prospective, Randomized Controlled Study.

Negative pressure wound therapy (NPWT) is a technique using vacuum dressing to promote wound healing in complicated wound. However, for many patients, the application and removal of the NPWT is source of procedural pain. The authors hypothesized that administering cold sterile water into the NPWT sponge would decrease pain during dressing changes. A prospective randomized controlled study was conducted on 27 patients who were undergoing 81 NPWT wound dressing changes (n = 81) at a single institution between October 2016 and September 2017. Each patient had 3 NPWT dressing changes. Cold sterile water (5.74 °C), room temperature sterile water (26.89 °C), and nothing were randomized and administered in the NPWT tubing into the sponge 10 minutes before changing the dressing in each and every procedure. Pain scores were assessed using a 0 to 10 numeric pain scale. Patients administered with cold water reported less pain than those administered with room temperature sterile water during the dressing change (4 vs 5.67; P < .003), and much less pain than those with nothing instilled before dressing change (4 vs 6.59; P < .001). There is no statistically significant difference in pain score between using the room temperature sterile water group and the control group that instilled nothing (5.67 vs 6.59; P = .065). This study has shown that cold water administered through the suction tubing before the dressing change had a better reduction in pain score than using room temperature sterile water and the control group.

A Cadaveric Study of Dye Spreading: Determining the Ideal Injection Pattern for Masseter Hypertrophy.

BACKGROUND: Masseter hypertrophy is the main cause of an asymmetrical and squared lower facial contour in the Asian community. Botulinum toxin injection technique is crucial to treat this condition. OBJECTIVE: To improve injection techniques for masseter hypertrophy by elucidating the distribution of the injections within the masseter. METHODS: Thirty masseter muscles were divided into 6 groups of 5 muscles each. Each group received one 0.2- or 0.3-mL injection at Point A, B, or C according to a three-point technique. Muscle dimensions and dye of the primary and secondary dye spreading were measured. RESULTS: The average muscle length, width, and thickness were 69.87, 33.50, and 11.23 mm, respectively. The average primary longitudinal and horizontal spreading was 36.56 and 15.60 mm, respectively. No statistically significant difference was found between 0.2- and 0.3-mL injections at each point. CONCLUSION: The three-point technique best fits in the safe zone and should be the standard injection technique for masseter hypertrophy. Injection at Points B and C may create secondary spreading that affect the risorius muscle and the parotid gland which are the cause of asymmetrical smiling and xerostomia, respectively. The dosage should be adjusted according to the muscle volume and not only the thickness.

The Anatomical Study of the Nasal Septal Cartilage with its Clinical Implications.

BACKGROUND: Several rhinoplasty and nasal reconstruction procedures require cartilage. Various studies have reported on the nasal septal cartilage as a donor site for Caucasian and Asian populations. However, studies regarding the Thai nasal septal cartilage dimensions are rare. This study aimed to examine the length, height, area, and thickness of the nasal septal cartilage, along with implications of the size and quantity of the available cartilage, for grafting in Thai cadavers. METHODS: We analyzed the nasal septal cartilage in 42 Thai cadavers. The length, height, area, and thickness were digitally measured using ImageJ 1.52 software, along with the size and area of the available cartilage for grafting after preserving a 10-mm L-strut. Data were compared between sexes. RESULTS: The mean height, length, and area of the nasal septal cartilage were 30.96 ± 5.90 mm, 26.13 ± 6.90 mm, and 636.10 ± 196.13 mm2, respectively. The length did not differ significantly between sexes. However, the height and area in male cadavers were greater than those in female cadavers. The mean thickness ranged from 0.77 to 1.02 mm depending on the area, with the thickest and thinnest areas being the superoposterior and inferoposterior parts, respectively. The mean height, length, and area of the harvestable cartilage were 20.96 mm, 16.13 mm, and 384.84 mm2, respectively, after excluding the L-strut. CONCLUSION: Our results provide major anatomical data of the Thai nasal septal cartilage. Its implication for use of the septal cartilage for grafting is 21 mm in height and 16 mm in length, which is sufficiently safe to maintain nasal support. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these evidence-based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.