Reflectance confocal microscopy and dermoscopy aid in evaluating repigmentation within or adjacent to lentigo maligna melanoma surgical scars.

BACKGROUND: Determining whether repigmentation within or adjacent to lentigo maligna or lentigo maligna melanoma (LM/LMM) scars represents recurrence of melanoma is challenging. The use of reflectance confocal microscopy (RCM) and dermoscopy may aid in differentiating true melanoma recurrence from other causes of repigmentation. OBJECTIVES: To describe the characteristics of repigmentation within or adjacent to LM/LMM scars observable on RCM and dermoscopy. METHODS: We retrospectively analysed patients who presented with new pigmentation within or adjacent to scars from surgically treated LM/LMM between January 2014 and December 2018. Clinical and demographic characteristics and time to recurrence were recorded. RCM was used to evaluate areas of pigmentation before biopsy. If available, dermoscopic images were also evaluated. RESULTS: In total, 30 confocal studies in 29 patients were included in the study cohort. Twenty-one patients had biopsy-confirmed recurrent LM/LMM; the remainder had pigmented actinic keratosis (n = 4) or hyperpigmentation/solar lentigo (n = 5). RCM had sensitivity of 95.24% (95% CI, 76.18-99.88%), specificity of 77.7% (95% CI, 39.99-97.19%), positive predictive value of 90.91% (95% CI, 74.58-97.15%) and negative predictive value of 87.5% (95% CI, 50.04-98.0%). The most common dermoscopic feature observed among patients with recurrent LM/LMM was focal homogeneous or structureless areas of light-brown pigmentation (92.8% vs. 37.5% in patients with other diagnoses; P = 0.009). LM-specific dermoscopic criteria were present in only 28.5% of patients with recurrent LM/LMM. CONCLUSIONS: Reflectance confocal microscopy and dermoscopy are valuable tools for the comprehensive evaluation of repigmentation within or adjacent to LM scars.

Incompletely excised lentigo maligna melanoma is associated with unpredictable residual disease: clinical features and the emerging role of reflectance confocal microscopy.

BACKGROUND: Lentigo maligna/lentigo maligna melanoma (LM/LMM) poses a treatment and surgical challenge given unpredictable subclinical extension resulting in incomplete excision. OBJECTIVES: To describe the demographic, clinical and pathologic characteristics of incompletely excised LM/LMM. To evaluate the potential role of reflectance confocal microscopy (RCM). PATIENTS AND METHODS: A retrospective review of a melanoma database at a tertiary cancer centre for patients referred with 'incompletely excised LM/LMM' or 'incompletely excised melanoma' between October 2006 and July 2017. We recorded clinical and pathological data and surgical margins needed to clear the residual LM/LMM. The second part consisted of a prospective cohort of patients in which RCM was performed when presenting with incompletely excised LM/LMM. RESULTS: We included a total of 67 patients (retrospective + prospective cohort); mean age was 64.9 (standard deviation: 11.3) years and 52.2% were males. For the retrospective cohort (n = 53), the mean scar size was 3.4 cm. The average initial margins excised prior to presentation were 4.8 mm (range 3-7 mm). The average additional margin needed to clear the residual, incompletely excised LM/LMM was 7.8 mm. For the prospective cohort (n = 14), there were no differences in age, gender or size when compared to the retrospective cohort. RCM had a diagnostic accuracy of 78.6%, a sensitivity of 90.9%, a specificity of 33.3% and a positive predictive value of 83.3% for the detection of incompletely excised LM/LMM. CONCLUSIONS: Incompletely excised LM/LMM is a poorly characterized clinical-pathological scenario that may require considerable extra margins for microscopic clearance. RCM may emerge as a valuable tool for the evaluation of patients with incompletely excised LM/LMM.

Peri-operative delineation of non-melanoma skin cancer margins in vivo with handheld reflectance confocal microscopy and video-mosaicking.

BACKGROUND: The surgical removal of non-melanoma skin cancers (NMSCs) is guided by the pathologic examination of margins. However, the preparation of histopathology is time consuming, labour-intensive and requires separate laboratory infrastructure. Furthermore, when histopathology indicates positive margins, patients must return for re-excisions. Reflectance confocal microscopy (RCM) with a new video-mosaicking approach can noninvasively delineate margins directly on patients and potentially guide surgery in real-time, augmenting the traditional approaches of histopathology. OBJECTIVE: To assess a new peri-operative RCM video-mosaicking approach for comprehensive delineation of NMSC margins on patients in vivo. METHODS: Thirty-five patients undergoing Mohs micrographic surgery (MMS) in the Mohs surgery unit at Memorial Sloan Kettering Cancer Center, New York, NY were included in the study. RCM imaging was performed before and after the first staged excision by acquiring videos along the surgical margins (epidermal, peripheral and deep dermal) of each wound, which were subsequently processed into video-mosaics. Two RCM evaluators read and assessed video-mosaics, and subsequently compared to the corresponding Mohs frozen histopathology. RESULTS: Reflectance confocal microscopy videos and video-mosaics displayed acceptable imaging quality (resolution and contrast), pre-operatively in 32/35 (91%) NMSC lesions and intra-operatively in 29/35 lesions (83%). Pre-operative delineation of margins correlated with the histopathology in 32/35 (91%) lesions. Intra-operative delineation correlated in 10/14 (71%) lesions for the presence of residual tumour and in 18/21 (86%) lesions for absence. Sensitivity/specificity were 71%/86% and 86%/81% for two RCM video-mosaic evaluators, and overall agreement was 80% and 83% with histopathology, with moderate inter-evaluator agreement (k = 0.59, P ≤ 0.0002). CONCLUSIONS: Peri-operative RCM video-mosaicking of NMSC margins directly on patients may potentially guide surgery in real-time, serve as an adjunct to histopathology, reduce time spent in clinic and reduce the need for re-excisions. Further testing in larger studies is needed.

Subtyping cognitive profiles in Autism Spectrum Disorder using a Functional Random Forest algorithm.

DSM-5 Autism Spectrum Disorder (ASD) comprises a set of neurodevelopmental disorders characterized by deficits in social communication and interaction and repetitive behaviors or restricted interests, and may both affect and be affected by multiple cognitive mechanisms. This study attempts to identify and characterize cognitive subtypes within the ASD population using our Functional Random Forest (FRF) machine learning classification model. This model trained a traditional random forest model on measures from seven tasks that reflect multiple levels of information processing. 47 ASD diagnosed and 58 typically developing (TD) children between the ages of 9 and 13 participated in this study. Our RF model was 72.7% accurate, with 80.7% specificity and 63.1% sensitivity. Using the random forest model, the FRF then measures the proximity of each subject to every other subject, generating a distance matrix between participants. This matrix is then used in a community detection algorithm to identify subgroups within the ASD and TD groups, and revealed 3 ASD and 4 TD putative subgroups with unique behavioral profiles. We then examined differences in functional brain systems between diagnostic groups and putative subgroups using resting-state functional connectivity magnetic resonance imaging (rsfcMRI). Chi-square tests revealed a significantly greater number of between group differences (p < .05) within the cingulo-opercular, visual, and default systems as well as differences in inter-system connections in the somato-motor, dorsal attention, and subcortical systems. Many of these differences were primarily driven by specific subgroups suggesting that our method could potentially parse the variation in brain mechanisms affected by ASD.

[Therapeutic potential of Cannabis sativa]. / Potencial uso terapéutico de cannabis.

Cannabis sativa (marihuana) is considered an illicit drug due to its psychoactive properties. Recently, the Chilean government opened to the use cannabis in the symptomatic treatment of some patients. The biological effects of cannabis render it useful for the complementary treatment of specific clinical situations such as chronic pain. We retrieved scientific information about the analgesic properties of cannabis, using it as a safe drug. The drug may block or inhibit the transmission of nervous impulses at different levels, an effect associated with pain control. Within this context and using adequate doses, forms and administration pathways, it can be used for chronic pain management, considering its effectiveness and low cost. It could also be considered as an alternative in patients receiving prolonged analgesic therapies with multiple adverse effects.

Carbon dioxide laser ablation of basal cell carcinoma with visual guidance by reflectance confocal microscopy: a proof-of-principle pilot study.

BACKGROUND: Laser ablation is an alternative, nonsurgical treatment modality for low-risk basal cell carcinoma (BCC). However, lack of confirmative tumour destruction or residual tumour presence has been a limiting factor to its adoption. Reflectance confocal microscopy (RCM) provides noninvasive, cellular-level resolution imaging of the skin and is capable of identifying tumour. OBJECTIVES: To evaluate the use of RCM to guide carbon dioxide (CO2 ) laser ablation of BCC, confirm destruction and correlate findings with histology. METHODS: RCM was used preablation to evaluate for features of BCC. Ablation was performed with a CO2 laser, and the response rapidly assessed using handheld RCM to evaluate for residual tumour. Confirmative pathology was used to verify confocal imaging. RESULTS: Preablation RCM imaging identified tumour with features not identified on normal, surrounding skin. Postablation, RCM documented complete removal of tumour in six cases and residual tumour in two. Histological examination identified the ablated area and confirmed clearance of tumour in the six aforementioned cases and corroborated confocal findings for residual tumour in the other two cases. CONCLUSIONS: We report successful treatment of superficial and nodular BCC using CO2 laser ablation augmented by RCM imaging for preablation guidance and verification of tumour removal postablation. Akin to complete circumferential and deep margin control techniques, using RCM helps to map peripheral and deep BCC margins to hone in on areas exhibiting persistent tumour after ablation. CO2 laser ablation visually guided by RCM can help circumvent previously cited limiting factors of laser ablation for tumour destruction by providing cellular-level resolution imaging of tumour and margin assessment in between each laser pass and postablation.

Role of the Streptococcus mutans CRISPR-Cas systems in immunity and cell physiology.

CRISPR-Cas systems provide adaptive microbial immunity against invading viruses and plasmids. The cariogenic bacterium Streptococcus mutans UA159 has two CRISPR-Cas systems: CRISPR1 (type II-A) and CRISPR2 (type I-C) with several spacers from both CRISPR cassettes matching sequences of phage M102 or genomic sequences of other S. mutans. The deletion of the cas genes of CRISPR1 (ΔC1S), CRISPR2 (ΔC2E), or both CRISPR1+2 (ΔC1SC2E) or the removal of spacers 2 and 3 (ΔCR1SP13E) in S. mutans UA159 did not affect phage sensitivity when challenged with virulent phage M102. Using plasmid transformation experiments, we demonstrated that the CRISPR1-Cas system inhibits transformation of S. mutans by the plasmids matching the spacers 2 and 3. Functional analysis of the cas deletion mutants revealed that in addition to a role in plasmid targeting, both CRISPR systems also contribute to the regulation of bacterial physiology in S. mutans. Compared to wild-type cells, the ΔC1S strain displayed diminished growth under cell membrane and oxidative stress, enhanced growth under low pH, and had reduced survival under heat shock and DNA-damaging conditions, whereas the ΔC2E strain exhibited increased sensitivity to heat shock. Transcriptional analysis revealed that the two-component signal transduction system VicR/K differentially modulates expression of cas genes within CRISPR-Cas systems, suggesting that VicR/K might coordinate the expression of two CRISPR-Cas systems. Collectively, we provide in vivo evidence that the type II-A CRISPR-Cas system of S. mutans may be targeted to manipulate its stress response and to influence the host to control the uptake and dissemination of antibiotic resistance genes.

Melanocytic naevi with globular and reticular dermoscopic patterns display distinct BRAF V600E expression profiles and histopathological patterns.

BACKGROUND: BRAF (v-raf murine sarcoma viral oncogene homologue B) V600E mutations have been detected with high frequency in melanocytic naevi. Few studies have stratified analyses by naevus dermoscopic pattern. OBJECTIVES: To determine the frequency of BRAF V600E expression and histopathological pattern in acquired melanocytic naevi distinguished by a globular vs. reticular dermoscopic pattern. METHODS: We retrospectively identified histologically proven melanocytic naevi with banal reticular or globular dermoscopic patterns and evaluated BRAF V600E expression using immunohistochemistry. RESULTS: BRAF V600E expression was detected in 11 of 12 globular naevi vs. four of 13 reticular naevi (91·7% vs. 30·1%, P = 0·004). A predominantly dermal growth pattern (P < 0·001) and the presence of large junctional nests (P = 0·017) were each associated with a globular dermoscopic pattern. The presence of either a predominantly dermal growth pattern or large junctional nests was found in 13 of 15 naevi positive for BRAF V600E and in two of 10 naevi negative for BRAF V600E (86·7% vs. 20%, P = 0·002). CONCLUSIONS: The frequency of BRAF V600E mutations differs in naevi distinguished by unique dermoscopic structures and microanatomical growth patterns. Globular naevi, which most often histologically correspond to a predominantly dermal growth pattern and/or the presence of large junctional nests, are significantly more likely to express BRAF V600E than reticular naevi. These preliminary results require validation, but may directly inform future studies of naevogenesis and melanoma genesis.

Regulation of bacteriocin production and cell death by the VicRK signaling system in Streptococcus mutans.

The VicRK two-component signaling system modulates biofilm formation, genetic competence, and stress tolerance in Streptococcus mutans. We show here that the VicRK modulates bacteriocin production and cell viability, in part by direct modulation of competence-stimulating peptide (CSP) production in S. mutans. Global transcriptome and real-time transcriptional analysis of the VicK-deficient mutant (SmuvicK) revealed significant modulation of several bacteriocin-related loci, including nlmAB, nlmC, and nlmD (P < 0.001), suggesting a role for the VicRK in producing mutacins IV, V, and VI. Bacteriocin overlay assays revealed an altered ability of the vic mutants to kill related species. Since a well-conserved VicR binding site (TGTWAH-N(5)-TGTWAH) was identified within the comC coding region, we confirmed VicR binding to this sequence using DNA footprinting. Overexpression of the vic operon caused growth-phase-dependent repression of comC, comDE, and comX. In the vic mutants, transcription of nlmC/cipB encoding mutacin V, previously linked to CSP-dependent cell lysis, as well as expression of its putative immunity factor encoded by immB, were significantly affected relative to the wild type (P < 0.05). In contrast to previous reports that proposed a hyper-resistant phenotype for the VicK mutant in cell viability, the release of extracellular genomic DNA was significantly enhanced in SmuvicK (P < 0.05), likely as a result of increased autolysis compared with the parent. The drastic influence of VicRK on cell viability was also demonstrated using vic mutant biofilms. Taken together, we have identified a novel regulatory link between the VicRK and ComDE systems to modulate bacteriocin production and cell viability of S. mutans.

Phenolic profile and antioxidant activity of Euonymus japonicus Thunb.

The aim of this study was to characterise the phenolic profile and evaluate the antioxidant activity of aqueous and methanol leaf extracts from Euonymus japonicus Thunb., a species of flowering plant in the Celastraceae family. Liquid chromatography (HPLC) with diode array and electrospray-tandem mass spectrometry (DAD-ESI-MSn) detection was used to characterise and quantify phenolic compounds. Thirty-two compounds were detected; among them, flavonoids (mainly quercetin and kaempferol glycosides) were the most abundant compounds in both extracts. The results obtained for DPPH and ABTS assays in the extracts were in agreement with the highest content of phenolics in the methanol extract.

Investigation by ICP-MS of trace element levels in vegetable edible oils produced in Spain.

The content of trace elements (Ag, As, Ba, Be, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Ti, Tl and V) in edible oils (virgin olive, olive, pomace-olive, sunflower, soybean and corn) from Spain was determined, using inductively-coupled plasma-mass spectrometry (ICP-MS) after microwave digestion, employing only nitric acid in this step. The method has been validated by using both an oil reference material and recovery experiments over different oil samples, obtaining satisfactory results in all cases. Inter-day repeatabilities lower than 10% were observed for all of the analysed elements in the different kinds of oil samples. Studying the content of trace elements, in order to detect tendencies in the samples of the same type of oil, principal components analysis was used. Promising groupings were observed using a model with two principal components and retaining 75.3% of the variance.

Sensitive determination of indomethacin in pharmaceuticals and urine by sequential injection analysis and optosensing.

A selective and sensitive method based on coupling of sequential injection analysis (SIA) and optosensing was developed and applied to the determination of indomethacin in pharmaceutical and urinary samples. After alkaline hydrolysis, the fluorescent product generated from indomethacin is inserted in the flow system, transitorily retained on an active solid support (Sephadex QAE A-25) filling the flowcell, and monitored at 283/371 nm (lamda ex / lamda em). The system was calibrated for two sample volumes, 100 and 1000 microL. It showed a linear dynamic range of 0.5-6.5 ng/mL, with an LOD of 0.15 ng/mL and an RSD of 3.9% (n=10) when the highest sample volume was used. The proposed fluorometric SIA optosensor was applied to the determination of indomethacin in both pharmaceuticals and urine samples, and satisfactory results were obtained.

Comparative study of the phytochemical and mineral composition of fresh and cooked broccolini.

Broccolini is originated from crossing the regular broccoli with Chinese kale. Consequently, it has similar properties to these vegetables, but other very particular characteristics. Its consumption has increased in the last few years and, consequently, there have been some studies related to its quality parameters and the influence of different cooking methods. Nevertheless, changes on its phenolic composition and mineral content originated by its cooking have not been investigated in-depth so far. Here we report the phytochemical profile of broccolini before and after boiling, steaming, and griddling cooking treatments. The mineral content and phytochemicals were assessed by inductively coupled plasma-mass spectrometry (ICP-MS) and high performance liquid chromatography-mass spectrometry (HPLC-MS), respectively. The main phenolics (mainly hydroxycinnamic acid derivatives from caffeic, coumaric, ferulic and sinapic acids) were quantified. Three oxylipins, three flavonoid glycosides and the glucosinolate glucobrassicin were also identified. ABTS and DPPH assays were also used as screening methods to assess the antioxidant potential of broccolini. A significant loss of the phenolic compounds and a reduction of the antioxidant activity were observed after the three cooking methods. Clear disadvantages were detected when broccolini was boiled, namely high losses of phenolic acids and derivatives (70%). Steaming and griddling also led to a significant loss of phenolics (50%) from fresh broccolini. The mineral content of this vegetable after domestic cooking procedures is also reported for the first time, calculating the contribution of broccolini consumption to official daily recommendations.

Correction: Role of growth arrest-specific gene 6-mer axis in multiple myeloma.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Development of an automated chemiluminescence flow-through sensor for the determination of 5-aminosalicylic acid in pharmaceuticals: a comparative study between sequential and multicommutated flow techniques.

This work is aimed at demonstrating the potential of the implementation of automatic flow systems in optosensors using chemiluminescence detection. With this purpose, two automatic methodologies, multicommutation and sequential injection analysis (SIA), have been applied to the analysis of 5-aminosalicylic acid (ASA). The analyte is determined for the first time making use of its chemiluminescence reaction with permanganate anion, previously immobilized on an appropriate solid support in the detection area. First, the study of the most appropriate commercial flow-through cell and the optimum conditions for the reaction were performed. Second, the main differences in terms of flow variables and analytical parameters for multicommutation and SIA approaches were stated. Both methodologies were applied to the determination of the analyte in pharmaceuticals obtaining satisfactory results. Finally, the advantages and disadvantages of both proposed methods and the recoveries obtained from pharmaceuticals were statistically compared.

Phenolic profile and antioxidant activity of Jasonia glutinosa herbal tea. Influence of simulated gastrointestinal in vitro digestion.

In order to value J. glutinosa DC (rock tea), we characterised its phenolic profile and antioxidant activity. The study was performed in aqueous extracts before and after a simulated in vitro digestion to obtain data regarding phenolics bioavailability. Methanolic extracts were also analysed for comparison purposes. Phytochemical profiles were determined by high-performance liquid chromatography with mass spectrometric detection, whereas total phenolic content (TPC) and antioxidant assays were performed by conventional spectrophotometric methods. The most abundant compounds were dicaffeoylquinic acids, representing more than 90% of phenolics in tea infusions. Statistically significant differences were observed for all parameters except for TPC in methanol and aqueous extracts. Both phenolics amount and antioxidant activities were lower after the in vitro digestion of the infusions. However, although phenolics were lost during the simulated digestion, rock tea is still a good source of bioactive compounds with potential applications in the pharmaceutical or nutraceutical industries.