Visualising cancer in 3D: 3-Dimensional Tissue Imaging for management of cutaneous basal cell carcinoma.

Surgical management of basal cell carcinoma (BCC) typically involves surgical excision with post-operative margin assessment using the bread-loafing technique; or gold-standard Mohs micrographic surgery (MMS), where margins are iteratively examined for residual cancer after tumour removal, with additional excisions performed upon detecting residual tumour at margins. There is limited sampling of resection margins with bread loafing, with detection of positive margins 44% of the time using 2 mm intervals. To resolve this, we have developed three-dimensional (3D) Tissue Imaging for: (1) complete examination of cancer margins and (2) detection of tumour proximity to nerves and blood vessels. 3D Tissue optical clearing with a light sheet imaging protocol was developed for margin assessment in two datasets assessed by two independent evaluators: (1) 48 samples from 29 patients with varied BCC subtypes, sizes and pigmentation levels; (2) 32 samples with matching Mohs' surgeon reading of tumour margins using two-dimensional haematoxylin & eosin-stained sections. The 3D Tissue Imaging protocol permits a complete examination of deeper and peripheral margins. Two independent evaluators achieved negative predictive values of 92.3% and 88.24% with 3D Tissue Imaging. Images obtained from 3D Tissue Imaging recapitulates histological features of BCC, such as nuclear crowding, palisading and retraction clefting and provides a 3D context for recognising normal skin adnexal structures. Concurrent immunofluorescence labelling of nerves and blood vessels allows visualisation of structures closer to tumour-positive regions, which may have a higher risk for neural and vascular infiltration. Together, this method provides more information in a 3D spatial context, enabling better cancer management by clinicians.

Regulating the transcriptomes that mediate the conversion of fibroblasts to various nervous system neural cell types.

Our understanding of the mechanism of cell fate transition during the direct reprogramming of fibroblasts into various central nervous system (CNS) neural cell types has been limited by the lack of a comprehensive analysis on generated cells, independently and in comparison with other CNS neural cell types. Here, we applied an integrative approach on 18 independent high throughput expression data sets to gain insight into the regulation of the transcriptome during the conversion of fibroblasts into induced neural stem cells, induced neurons (iNs), induced astrocytes, and induced oligodendrocyte progenitor cells (iOPCs). We found common down-regulated genes to be mostly related to fibroblast-specific functions, and suggest their potential as markers for screening of the silencing of the fibroblast-specific program. For example, Tagln was significantly down-regulated across all considered data sets. In addition, we identified specific profiles of up-regulated genes for each CNS neural cell types, which could be potential markers for maturation and efficiency screenings. Furthermore, we identified the main TFs involved in the regulation of the gene expression program during direct reprogramming. For example, in the generation of iNs from fibroblasts, the Rest TF was the main regulator of this reprogramming. In summary, our computational approach for meta-analyzing independent expression data sets provides significant details regarding the molecular mechanisms underlying the regulation of the gene expression program, and also suggests potentially useful candidate genes for screening down-regulation of fibroblast gene expression profile, maturation, and efficiency, as well as candidate TFs for increasing the efficiency of the reprogramming process.

Common Gene Expression Patterns in the Generation of Induced Neurons From Fibroblasts.

In the current study, we analyzed ten gene expression data sets including RNA-sequencing and microarray experiment data during the direct reprogramming of mouse and human fibroblasts to induced neurons and found common gene expression pattern across all data sets for this conversion.

Meta-Analysis of Transcriptome Regulation During Induction to Cardiac Myocyte Fate From Mouse and Human Fibroblasts.

Ectopic expression of a defined set of transcription factors (TFs) can directly convert fibroblasts into a cardiac myocyte cell fate. Beside inefficiency in generating induced cardiomyocytes (iCMs), the molecular mechanisms that regulate this process remained to be well defined. The main purpose of this study was to provide better insight on the transcriptome regulation and to introduce a new strategy for candidating TFs for the transdifferentiation process. Eight mouse and three human high quality microarray data sets were analyzed to find differentially expressed genes (DEGs), which we integrated with TF-binding sites and protein-protein interactions to construct gene regulatory and protein-protein interaction networks. Topological and biological analyses of constructed gene networks revealed the main regulators and most affected biological processes. The DEGs could be categorized into two distinct groups, first, up-regulated genes that are mainly involved in cardiac-specific processes and second, down-regulated genes that are mainly involved in fibroblast-specific functions. Gata4, Mef2a, Tbx5, Tead4 TFs were identified as main regulators of cardiac-specific gene expression program; and Trp53, E2f1, Myc, Sfpi1, Lmo2, and Meis1 were identified as TFs which mainly regulate the expression of fibroblast-specific genes. Furthermore, we compared gene expression profiles and identified TFs between mouse and human to find the similarities and differences. In summary, our strategy of meta-analyzing the data of high-throughput techniques by computational approaches, besides revealing the mechanisms involved in the regulation of the gene expression program, also suggests a new approach for increasing the efficiency of the direct reprogramming of fibroblasts into iCMs. J. Cell. Physiol. 232: 2053-2062, 2017. © 2016 Wiley Periodicals, Inc.

Comprehensive Dissection of Transcriptome Data and Regulatory Factors in Pancreatic Cancer Cells.

Features of pancreatic cancers include high mortality rates caused by rapid tumor progression and a lack of effective therapy. Underpinning the molecular mechanisms involved in the alteration of the gene expression program in the pancreatic cancer remains to be understood. In the current study, we performed a comprehensive analysis using 282 pancreatic tumor and normal samples from seven independent expression data sets to provide a better view on the interactions between different transcription factors (TFs) and the most affected biological pathways in pancreatic cancer. We highlighted common differentially expressed genes (DEGs) and common affected processes within pancreatic cancer samples. We revealed 16 main DE-TFs that regulated gene expression alterations as well as the most significant processes in pancreatic cancer compared to normal cells. For example, we found the upregulated FOXM1 to be a top regulator of pancreatic cellular transformation based on results from different analyses, including from its regulation of gene regulatory networks, its presence in protein complex, its significant regulation of genes related to cancer pathways, and its regulation of most of the identified DE-TFs. Furthermore, we provided a model and assessed the role of different DE-TFs in the regulation of the most affected pancreatic- and cancer-specific processes. In conclusion, our bioinformatics meta-analysis of high throughput expression data sets, besides clarifying common affected genes and pathways, also showed the mechanisms involved in regulating these common profiles. Our results, especially for DE-TFs, could potentially be useful for screening for pancreatic cancer, and for confirming or determining novel pharmacological targets. J. Cell. Biochem. 118: 3976-3985, 2017. © 2017 Wiley Periodicals, Inc.

Treatment of xanthelasma palpebrarum with a 1064-nm, Q-switched Nd:YAG laser.

BACKGROUND: Xanthelasma palpebrarum is the most common cutaneous xanthoma characterized by soft, yellow papules or plaques that arise on the periorbital skin. As these lesions can be cosmetically disfiguring, many patients seek medical help to remove these lesions. OBJECTIVE: To determine the effectiveness and minimum number of treatment sessions with a 1064-nm, Q-switched neodymium-doped yttrium aluminum garnet (Nd:YAG) laser for the treatment of xanthelasma. METHODS: A retrospective review of patients with xanthelasma consecutively treated with Q-switched Nd:YAG laser was conducted. Forty-six patients with 103 lesions were identified from January 2012 through August 2015. Photographs taken of patients immediately before treatment and 4-8 weeks after treatment were independently evaluated by 2 dermatologists. RESULTS: After a single treatment session, 93.2% of lesions had some degree of clearance. All lesions had excellent-to-complete clearance after at least 4 treatment sessions. Patients usually required 4 treatment sessions for optimal results. LIMITATIONS: This was a retrospective study. Treatment parameters varied, follow-up periods were not uniform, and response was not assessed with a validated scale. CONCLUSION: The Q-switched Nd:YAG laser is effective and well tolerated in the treatment of xanthelasma in our study population.

Treatment of Facial Acne Scarring With Fractional Carbon Dioxide Laser in Asians, a Retrospective Analysis of Efficacy and Complications.

BACKGROUND: Fractional carbon dioxide (CO2) laser resurfacing unites the idea of fractional photothermolysis with an ablative 10,600-nm wavelength. This technology permits effective treatment of acne scarring, with shorter recovery and a decreased side effect profile as compared to traditional CO2 laser resurfacing. OBJECTIVE: The aims of this study were to study the efficacy and safety of fractional CO2 laser for acne scars in Asian patients. METHODS: This was a retrospective study of all patients treated with the fractional CO2 laser for facial acne scarring between January 2, 2008, and December 31, 2010, at the National Skin Centre, Singapore. Data reviewed included sex, age, and severity of acne, type of scarring, previous treatment history, frequency of treatments, adverse events, and efficacy. RESULTS: Two hundred and ten fractional CO2 laser treatments for facial acne scars were performed on 107 patients (65 men and 42 women, Fitzpatrick skin Types II to V) during the study period. Sixteen of 107 patients experienced (15.0%) adverse events. The adverse events include hyperpigmentation (6.4%), blistering (4.0%), crusting (2.9%), aggravation of inflammatory acne lesions (1.7%), and scarring (0.6%). There were no reported side effects of hypopigmentation, bacterial or viral infection. Follow-up results after final laser treatment showed that 66.4% of patients reported Grade 1 skin texture improvement, that is, <25% (n = 71); 30.0% had Grade 2 improvement, that is, 25% to 50% improvement (n = 31); 3.7% had Grade 3 improvement, that is, 51% to 75% improvement (n = 4); and 0.9% had Grade 4 improvement, that is, >75% improvement (n = 1). CONCLUSION: The study demonstrated the efficacy and safety of a fractional CO2 laser in the treatment of acne scars in Asian. Future studies are required to establish optimum treatment parameters and achieve better clinical results.

Reflectance confocal microscopy is a useful non-invasive tool in the in vivo diagnosis of pigmented basal cell carcinoma in Asians.

BACKGROUND: The clinical differentiation between pigmented basal cell carcinoma (BCC) and other benign pigmented skin lesions can be challenging even with an additional dermoscopic evaluation, especially if the lesion is small. In vivo reflectance confocal microscopy (RCM) is an emerging, non-invasive imaging tool that allows near-microscopic evaluation of skin lesions. The features of RCM for pigmented BCC and seborrhoeic keratosis have previously been described. However, the use of RCM to differentiate between these clinically and dermoscopically challenging pigmented skin lesions among Asians has not yet been demonstrated. OBJECTIVES: We aimed to evaluate the usefulness of non-invasive RCM to differentiate between clinically and dermoscopically challenging pigmented skin lesions among Asians in a series of 11 lesions. METHODS: Nine patients with 11 clinically and dermoscopically difficult to distinguish pigmented skin lesions were evaluated by RCM to differentiate between pigmented BCC and benign lesions. In all cases, a histological confirmation of the RCM diagnosis was obtained. RESULTS: The clinical or dermoscopical characteristics were non-specific in all 10 cases. RCM detected features of pigmented BCC in nine patients and seborrhoeic keratosis in one patient. These were all confirmed by histological examination. CONCLUSIONS: This case series shows the value of non-invasive in vivo RCM imaging in the differentiation of malignant and benign pigmented lesions. Early diagnosis of small, pigmented BCC allows earlier excision with better prognosis. Future biopsies of benign lesions in cosmetic areas could also be avoided.

A retrospective review of cutaneous metastases at the National Skin Centre Singapore.

BACKGROUND/OBJECTIVES: Cutaneous metastases occur in up to 10% of all visceral malignancies. This study aims to determine the clinico-epidemiological characteristics of patients diagnosed with cutaneous metastases at a leading Asian tertiary dermatological centre. METHODS: A retrospective chart review was carried out on all patients diagnosed with cutaneous metastases from 2001 to 2010, identified through the National Skin Centre's clinical and histological databases. RESULTS: In all, 35 patients were diagnosed with cutaneous metastases at a mean age of 65 years, with a female: male ratio of 3:2 and with most (97%) being Chinese. Cutaneous metastases were the first presentation of cancer in 34% of patients (n = 12). The two commonest primaries were breast (49%, n = 17) and lung (9%, n = 3). Most had cutaneous metastases on the chest (37%, n = 13) followed by the pelvis (17%, n = 6) and 20% (n = 7) had skin lesions on multiple sites. Clinically, 46% of patients (n = 16) presented with nodule(s), 26% (n = 9) with plaque(s) and 20% (n = 7) with heterogeneous morphologies. Cutaneous metastases were suspected in 72% of patients (n = 25), attributed to clinical features (46%, n = 16) or the presence of active metastatic disease elsewhere (26%, n = 9). All lesions appeared dermal in origin. CONCLUSIONS: Cutaneous metastases are highly variable in presentation and a high index of suspicion is required for prompt diagnosis, especially in patients with a history of cancer, regardless of stage of treatment of the primary tumour.

Profile identification and characterization of risk perceptions and preventive behaviors during the COVID-19 pandemic: A latent profile analysis.

In a public health crisis, communication plays a vital role in making sure policies and recommendations from the government level get disseminated accurately to its people and is only considered as effective when the public accepts, supports, complies to, and engages in policies or behaves as per governments' recommendations. Adopting the multivariate audience segmentation strategy for health communication, this study uses a data-driven analytical method to (1) identify audience segments of public health crisis communication in Singapore based on knowledge, risk perception, emotional responses, and preventive behaviors; and (2) characterize each audience segment according to demographic factors, personality traits, information processing styles, and health information preferences. Results (N = 2033) from a web-based questionnaire executed in August 2021 have identified three audience segments: the less-concerned (n = 650), the risk-anxious (n = 142), and the risk-majority (n = 1,241). This study offers insights to how audiences of public health crisis communication perceive, process, and respond to information directed to them during the pandemic, thereby informing policy makers to tailor more targeted public health communication interventions in promoting positive attitude and behavior change.

Extramammary Paget disease: Immunohistochemistry is critical to distinguish potential mimickers.

Extra-mammary Paget disease (EMPD) is a rare intra-epithelial carcinoma that is usually found on the apocrine-rich skin of the perineum. We report 2 cases in which EMPD was initially misdiagnosed on the initial punch biopsy as melanoma-in-situ and Bowen disease respectively. Reasons for the misdiagnoses included a rare pigmented axillary variant of EMPD in the first case and atypical bowenoid features on H&E in the second. The cases are described with a critical review of the histopathological findings, along with a review of the current literature. This highlights the necessity of a comprehensive immunohistochemical panel for the assessment of intra-epithelial pagetoid atypical cells.

Synchronization of HeLa Cells to Various Interphases Including G1, S, and G2 Phases.

The typical cell cycle in eukaryotes is composed of four phases including the G1, S, G2, and M phases. G1, S, and G2 together are called interphase. Cell synchronization is a process that brings cultured cells at different stages of the cell cycle to the same phase, which allows the study of phase-specific cellular events. While interphase cells can be easily distinguished from mitotic cells by examining their chromosome morphology, it is much more difficult to separate and distinguish the interphases from each other. Here, we describe drug-derived protocols for synchronizing HeLa cells to various interphases of the cell cycle: G1 phase, S phase, and G2 phase. G1 phase synchronization is achieved through serum starvation, S phase synchronization is achieved through a double thymidine block, and G2 phase synchronization is achieved through the release of the double thymidine block followed by roscovitine treatment. Successful synchronization can be assessed using flow cytometry to examine the DNA content and Western blot to examine the expression of various cyclins.

Synchronization of HeLa Cells to Mitotic Subphases.

The cell cycle is a series of events leading to cell replication. When plated at low cell densities in serum-containing medium, cultured cells start to proliferate, moving through the four phases of the cell cycle: G1, S, G2, and M. Mitosis is the most dynamic period of the cell cycle, involving a major reorganization of virtually all cell components. Mitosis is further divided into prophase, prometaphase, metaphase, anaphase, and telophase, which can be easily distinguished from one another by protein markers and/or comparing their chromosome morphology under fluorescence microscope. The progression of the cell cycle through these mitotic subphases is tightly regulated by complicated molecular mechanisms. Synchronization of cells to the mitotic subphases is important for understanding these molecular mechanisms. Here, we describe a protocol to synchronize Hela cells to prometaphase, metaphase, and anaphase/telophase. In this protocol, Hela cells are first synchronized to the early S phase by a double thymidine block. Following the release of the block, the cells are treated with nocodazole, MG132, and blebbistatin to arrest them at prometaphase, metaphase, and anaphase/telophase, respectively. Successful synchronization is assessed using Western blot and fluorescence microscopy.

Melanoma in Singapore: A 20-year review of disease and treatment outcomes.

INTRODUCTION: Melanomas in Asians have different clinicopathological characteristics and prognosis from melanomas in Caucasians. This study reviewed the epidemiology and treatment outcomes of cutaneous melanoma diagnosed at a tertiary referral dermatology centre in Singapore, which has a multiracial population. The study also determined whether Asians had comparable relapse-free and overall survival periods to Caucasians in Singapore. METHOD: This is a retrospective review of cutaneous melanoma cases in our centre between 1996 and 2015. RESULTS: Sixty-two cases of melanoma were diagnosed in 61 patients: 72.6% occurred in Chinese, 19.4% in Caucasians and 3.2% in Indians, with an over-representation of Caucasians. Superficial spreading melanoma, acral lentiginous melanoma and nodular melanoma comprised 37.1%, 35.5% and 22.6% of the cases, respectively. The median time interval to diagnosis was longer in Asians than Caucasians; median Breslow's thickness in Asians were significantly thicker than in Caucasians (2.6mm versus 0.9mm, P=0.018) and Asians tend to present at a later stage. The mortality rates for Asians and Caucasians were 52% and 0%, respectively. CONCLUSION: More physician and patient education on skin cancer awareness is needed in our Asian-predominant population for better outcomes.

Efficacy and Safety of a New Cosmeceutical Regimen Based on the Combination of Snail Secretion Filtrate and Snail Egg Extract to Improve Signs of Skin Aging.

BACKGROUND: Two extracts derived from the gastropod Cryptomphalus aspersa have been shown to have dermal regeneration properties: SCA® (secretion filtrate) with fibroblast growth factor-like activity and IFC®-CAF (cellular activating factor), a snail egg extract with skin stem cell activation activity. OBJECTIVE: The objectives of this study were to evaluate the synergic antiaging activity and tolerability of SCA and IFC-CAF in a combined regimen compared to vehicle as a placebo control. METHODS: A three-month, single-center, double-blinded, randomized, vehicle-controlled trial assessed the effects of a daily skincare routine divided into two treatment phases, as follows: intensive (1 month) and maintenance (2 months). Fifty women, aged 45-65 years, with signs of photoaging were randomized to receive either the active ingredients (n=30) or vehicle (n=20). Clinical evaluations included objective measurements of barrier function and skin hydration, elasticity, and color/brightness. Subjective assessments were conducted according to the Rao-Goldman and Glogau scales for wrinkles, the Patient Global Assessment (PGA) scale and Investigator Global Assessment (IGA) scale. RESULTS: Subjects in the active treatment group experienced reductions in transepidermal water loss and significant improvements in skin roughness, firmness, and elasticity. Both groups showed significant improvements in fine lines and wrinkles. PGA and IGA assessments indicated greater improvement in the active treatment group. CONCLUSION: The active snail extract treatment appears to be effective in improving signs of skin aging in women 45 to 65 years old. Larger randomized, controlled studies are needed to confirm our results.

The Quadrivalent Human Papillomavirus Vaccine in Recalcitrant Non-genital Warts: A Retrospective Study.

INTRODUCTION: The human papillomavirus (HPV) vaccine has been reported to lead to clinical clearance of lesions when used as an off-label treatment for recalcitrant extragenital warts. The aim of the study is to evaluate the therapeutic and adverse effects of HPV vaccine as an adjunctive therapy for treatment-resistant acral warts. METHODS: Patients with persistent warts despite first and second line therapies, and subsequently receiving the quadrivalent HPV vaccine between July 2013 and June 2016 as an adjunctive treatment for recalcitrant warts at the National Skin Centre, were included. RESULTS: Twenty-six patients with a median age of 34 years (range 8 to 77 years) were treated with the HPV vaccine. Nineteen (73.1%) patients completed 3 doses of the vaccine, of whom 5 (26%) achieved complete clearance, 8 (42%) had partial clearance and 6 (32%) did not respond to the vaccine. Among the 4 patients who received 2 doses of the vaccine, 3 (75%) had complete clearance whereas 1 (25%) had partial improvement of their warts. None of the patients reported adverse reactions. CONCLUSION: Our study suggests a potential adjunctive role of the HPV vaccine in the treatment of acral warts recalcitrant to conventional therapy.

Optimisation of tube voltage range (kVp) for AP abdomen, pelvis and spine imaging of average patients with a digital radiography (DR) imaging system using a computer simulator.

OBJECTIVES: To investigate via computer simulation, an optimised tube voltage (kVp) range for caesium iodide (CsI)-based digital radiography (DR) of the abdomen, pelvis and lumbar spine. METHODS: Software capable of simulating abdomen, pelvis and spine radiographs was used. Five evaluators graded clinical image criteria in images of 20 patients at tube voltages ranging from 60 to 120 kVp in 10 kVp increments. These criteria were scored blindly against the same patient reconstructed at a specific reference kVp. Linear mixed effects analysis was used to evaluate image scores for each criterion and test for statistical significance. RESULTS: Score was dependent on tube voltage and image criteria; both were statistically significant. All criteria for all anatomies scored very poorly at 60 kVp. Scores for abdomen, pelvis and spine imaging peaked at 70, 70 and 100 kVp, respectively, but other kVp values were not significantly poorer. CONCLUSIONS: Results indicate optimum tube voltages of 70 kVp for abdomen and pelvis (with an optimum range 70-120 kVp), and 100 kVp (optimum range 80-120 kVp) for lumbar spine. ADVANCES IN KNOWLEDGE: There are no recommendations for optimised tube voltage parameters for DR abdomen, pelvis or lumbar spine imaging. This study has investigated and recommended an optimal tube voltage range.