Combinatorial Single-Cell Analyses of Granulocyte-Monocyte Progenitor Heterogeneity Reveals an Early Uni-potent Neutrophil Progenitor.

Granulocyte-monocyte progenitors (GMPs) have been previously defined for their potential to generate various myeloid progenies such as neutrophils and monocytes. Although studies have proposed lineage heterogeneity within GMPs, it is unclear if committed progenitors already exist among these progenitors and how they may behave differently during inflammation. By combining single-cell transcriptomic and proteomic analyses, we identified the early committed progenitor within the GMPs responsible for the strict production of neutrophils, which we designate as proNeu1. Our dissection of the GMP hierarchy led us to further identify a previously unknown intermediate proNeu2 population. Similar populations could be detected in human samples. proNeu1s, but not proNeu2s, selectively expanded during the early phase of sepsis at the expense of monocytes. Collectively, our findings help shape the neutrophil maturation trajectory roadmap and challenge the current definition of GMPs.

Skinny deeping: Uncovering immune cell behavior and function through imaging techniques.

As the largest organ of the body, the skin is a key barrier tissue with specialized structures where ongoing immune surveillance is critical for protecting the body from external insults. The innate immune system acts as first-responders in a coordinated manner to react to injury or infections, and recent developments in intravital imaging techniques have made it possible to delineate dynamic immune cell responses in a spatiotemporal manner. We review here key studies involved in understanding neutrophil, dendritic cell and macrophage behavior in skin and further discuss how this knowledge collectively highlights the importance of interactions and cellular functions in a systems biology manner. Furthermore, we will review emerging imaging technologies such as high-content proteomic screening, spatial transcriptomics and three-dimensional volumetric imaging and how these techniques can be integrated to provide a systems overview of the immune system that will further our current knowledge and lead to potential exciting discoveries in the upcoming decades.

A 3D-printed transepidermal microprojection array for human skin microbiome sampling.

Skin microbiome sampling is currently performed with tools such as swabs and tape strips to collect microbes from the skin surface. However, these conventional approaches may be unable to detect microbes deeper in the epidermis or in epidermal invaginations. We describe a sampling tool with a depth component, a transepidermal microprojection array (MPA), which captures microbial biomass from both the epidermal surface and deeper skin layers. We leveraged the rapid customizability of 3D printing to enable systematic optimization of MPA for human skin sampling. Evaluation of sampling efficacy on human scalp revealed the optimized MPA was comparable in sensitivity to swab and superior to tape strip, especially for nonstandard skin surfaces. We observed differences in species diversity, with the MPA detecting clinically relevant fungi more often than other approaches. This work delivers a tool in the complex field of skin microbiome sampling to potentially address gaps in our understanding of its role in health and disease.

Efficacy and safety of narrowband ultraviolet B versus combined ultraviolet A/narrowband ultraviolet B phototherapy in the treatment of chronic atopic dermatitis: A randomised double-blind study.

Phototherapy is a useful treatment modality for atopic dermatitis (AD). This is a prospective randomised double-blind study comparing the clinical efficacy of combined ultraviolet-A (UVA)/narrowband ultraviolet-B (NBUVB) versus NBUVB phototherapy in the treatment of chronic AD. Patients with moderate-to-severe AD were randomised to receive either UVA/NBUVB or NBUVB phototherapy twice weekly over 12 weeks. At baseline, weeks 6 and 12, Eczema Area And Severity Index (EASI), itch score and adverse effects were assessed. At baseline and week 12, disease-related quality of life was evaluated using the Dermatology Life Quality Index (DLQI). Nine patients were randomised to receive UVA/NBUVB and 10 received NBUVB. At week 12, both groups showed significant improvement in EASI and itch scores (p < 0.05). Significant improvement in DLQI was seen in the UVA/NBUVB arm (p = 0.009) with a trend towards improvement in the NBUVB arm (p = 0.11). The efficacy of both modalities were comparable, as were reported adverse effects aside from skin dryness which was higher in the NBUVB arm (40% vs. 0%, p = 0.033). Combined UVA/NBUVB and NBUVB phototherapy have comparable clinical efficacy and safety in the treatment of chronic AD. NBUVB may induce greater skin dryness.

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.

Silencing siRNA microneedle patches versus silicone sheets in reducing post-surgical scars: a randomized single-blinded intra-individually controlled clinical trial.

BACKGROUND: A common complication of wounds is exuberant growth of fibrotic scar tissue, which can lead to hypertrophic scars or keloids. There are currently no treatments with good evidence for preventing excessive scar tissue formation. In this study, we explored the use of microneedle patches containing siRNA inhibiting SPARC mRNA in reducing the volume of post-surgical scars. OBJECTIVE: We aim to compare the differences in the volume of post-surgical scars between daily application of siRNA-embedded dissolving microneedle patches and silicone sheets. The primary study outcome measure was the 3D volume of scar elevation.Our hypothesis was that scar formation in the half of the wound treated with siRNA microneedle patches will be lesser, as reflected by a smaller 3D volume, as compared to the half treated with silicone sheets. METHODS: This was an 8-week, single-blinded intra-individually controlled randomised trial in a tertiary dermatological centre. Patients with two-week-old post-operative wounds were recruited. Each half of the scar was randomly assigned to the microneedle patch or silicone sheet. Three-dimensional (3D) volumes were obtained from the scars via a high-resolution scanner at day 0, 30 and 60. RESULTS: At day 30, scars treated with the microneedle patches had a lower geometric mean volume of 0.79mm3 when compared to scars treated with silicone sheets, with a difference in mean percentage volume reduction of 10.70%.At day 60, scars treated with the microneedle patches had a statistically significant lower volume (8.88mm3) when compared to the side treated with silicone sheets (12.77mm3, p=0.005), with a difference in mean percentage reduction of 9.66%. Additionally, there was also a statistically significant difference between the percentage reduction in scar volume, compared to baseline, on the side treated with microneedle patches (mean=83.78%) compared to the side treated with silicone sheets (mean=74.11%). CONCLUSIONS: There was a significantly greater reduction in the volume of post-surgical scars on the side treated with microneedle patches compared to the side treated with silicone sheets. This demonstrates the use of transdermal gene silencing technology for scar inhibition and that siRNA microneedle patches can be an effective and safe modality in the reduction of scar tissue formation. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry (ANZCTR), ACTRN12622000558729, https://www.anzctr.org.au.

Clinical efficacy of a gentle anti-dandruff itch-relieving shampoo formulation.

Dandruff is a common scalp condition affecting almost half of the world's population. Despite its high prevalence, the exact pathophysiology is not well established and is understood to be multifactorial, with factors such as fungal colonization, sebaceous gland activity and individual factors being implicated. There is a need for an effective and safe shampoo that can target the above factors. Hence, we have developed a shampoo formulation with properties of oil control, moisturizing, non-irritative, anti-fungal, anti-microbial and itch-relieving. In this interventional, open-label study, we evaluated the efficacy and safety of this shampoo in reducing the clinical signs of dandruff and pruritus in patients with pre-existing mild-to-moderate dandruff over a course of 21-day treatment duration through self-assessment and objective clinical evaluations. After continued use of the shampoo, there was a significant decrease in the adherent and loose scalp flaking scores. Mean pruritus scores also decreased significantly across the 21-day time points. There were also no adverse events or skin intolerances reported. This study showed that our shampoo formulation has led to a significant reduction in both adherent and loose scalp flaking and pruritus when used in individuals suffering from mild to moderate dandruff. As such, it is an ideal shampoo, which can be used to effectively control dandruff.

Les pellicules sont une affection courante du cuir chevelu qui touche près de la moitié de la population mondiale. Malgré sa prévalence élevée, la physiopathologie exacte n'est pas bien établie et est comprise comme étant multifactorielle, avec des facteurs impliqués tels que la colonisation fongique, l'activité des glandes sébacées et des facteurs individuels. Un shampooing sûr et efficace pouvant cibler les facteurs évoqués ci-dessus est nécessaire. Par conséquent, nous avons développé une formulation de shampooing présentant des propriétés de contrôle du sébum, d'hydratation, non irritante, antifongique, antimicrobienne et de soulagement des démangeaisons. Dans cette essaie interventionnelle ouvert, nous avons évalué l'efficacité et la sécurité d'emploi de ce shampooing dans la réduction des signes cliniques de pellicules et de prurit chez des patients présentant des pellicules préexistantes légères à modérées sur une durée de traitement de 21 jours par le biais d'auto-évaluations et d'évaluations cliniques objectives. Après une utilisation continue du shampooing, une diminution significative des scores de squames adhérents et d'excoriation du cuir chevelu a été observée. Les scores de prurit moyens ont également significativement diminué tout au long de la période de 21 jours. De même, aucun événement indésirable ou intolérance cutanée n'a été rapporté. Cette étude a montré que notre formulation de shampooing a entraîné une réduction significative des squames adhérents, de l'excoriation du cuir chevelu et du prurit lorsqu'elle est utilisée chez des personnes souffrant de pellicules légères ou modérées. Il s'agit donc d'un shampooing idéal qui peut être utilisé pour contrôler efficacement les pellicules.

Vitamin E in Atopic Dermatitis: From Preclinical to Clinical Studies.

BACKGROUND: Oxidative stress and inflammation are some of the proposed mechanisms involved in the pathogenesis of atopic dermatitis (AD). Current pharmacotherapeutic approaches are effective yet they are not without adverse effects. Vitamin E has great potential as an adjunctive treatment for AD owing to its antioxidant and anti-inflammatory bioactivities. SUMMARY: This review article summarizes the current available evidence from cellular, animal and clinical studies on the relationship between vitamin E and AD. The future prospects of vitamin E are also discussed. Vitamin E in practice does not show any toxicity to humans within a range of reasonable dosage. Albeit rarely, vitamin E as a contact allergen should be considered. Collectively, this review envisaged vitamin E as an adjunctive treatment for AD patients. Future research on the distinct effects of different vitamin E isoforms as well as their delivery system in skin disorders is needed.

Bullous eosinophilic annular erythema.

Eosinophilic annular erythema is an idiopathic acute eosinophilic dermatosis. It is a rare condition, with approximately 30 cases reported in the English literature. It features annular, figurate urticarial edematous plaques primarily affecting the trunk and proximal limbs. During evaluation of a patient, secondary causes of eosinophilic inflammation such as allergy-related conditions (eczema, drug, urticaria, contact dermatitis), parasitic infestations, and autoimmune dermatoses will need to be excluded. We present an unusual case of a 47-year-old patient who developed this condition.

Reprint of "Multi-modal image cytometry approach - From dynamic to whole organ imaging".

Optical imaging is a valuable tool to visualise biological processes in the context of the tissue. Each imaging modality provides the biologist with different types of information - cell dynamics and migration over time can be tracked with time-lapse imaging (e.g. intra-vital imaging); an overview of whole tissues can be acquired using optical clearing in conjunction with light sheet microscopy; finer details such as cellular morphology and fine nerve tortuosity can be imaged at higher resolution using the confocal microscope. Multi-modal imaging combined with image cytometry - a form of quantitative analysis of image datasets - provides an objective basis for comparing between sample groups. Here, we provide an overview of technical aspects to look out for in an image cytometry workflow, and discuss issues related to sample preparation, image post-processing and analysis for intra-vital and whole organ imaging.

Techniques and Applications in Skin OCT Analysis.

The skin is the largest organ of our body. Skin disease abnormalities which occur within the skin layers are difficult to examine visually and often require biopsies to make a confirmation on a suspected condition. Such invasive methods are not well-accepted by children and women due to the possibility of scarring. Optical coherence tomography (OCT) is a non-invasive technique enabling in vivo examination of sub-surface skin tissue without the need for excision of tissue. However, one of the challenges in OCT imaging is the interpretation and analysis of OCT images. In this review, we discuss the various methodologies in skin layer segmentation and how it could potentially improve the management of skin diseases. We also present a review of works which use advanced machine learning techniques to achieve layers segmentation and detection of skin diseases. Lastly, current challenges in analysis and applications are also discussed.

The impact of ischemia-reperfusion injuries on skin resident murine dendritic cells.

Pressure ulcers are a chronic problem for patients or the elderly who require extended periods of bed rest. The formation of ulcers is due to repeated cycles of ischemia-reperfusion (IR), which initiates an inflammatory response. Advanced ulcers disrupt the skin barrier, resulting in further complications. To date, the immunological aspect of skin IR has been understudied, partly due to the complexity of the skin immune cells. Through a combination of mass cytometry, confocal imaging and intravital multiphoton imaging, this study establishes a workflow for multidimensionality single cell analysis of skin myeloid cell responses in the context of IR injury with high spatiotemporal resolution. The data generated has provided us with previously uncharacterized insights into the distinct cellular behavior of resident dendritic cells (DCs) and recruited neutrophils post IR. Of interest, we observed a drop in DDC numbers in the IR region, which was subsequently replenished 48h post IR. More importantly, in these cells, we observe an attenuated response to repeated injuries, which may have implications in the subsequent wound healing process.

Multi-modal image cytometry approach - From dynamic to whole organ imaging.

Optical imaging is a valuable tool to visualise biological processes in the context of the tissue. Each imaging modality provides the biologist with different types of information - cell dynamics and migration over time can be tracked with time-lapse imaging (e.g. intra-vital imaging); an overview of whole tissues can be acquired using optical clearing in conjunction with light sheet microscopy; finer details such as cellular morphology and fine nerve tortuosity can be imaged at higher resolution using the confocal microscope. Multi-modal imaging combined with image cytometry - a form of quantitative analysis of image datasets - provides an objective basis for comparing between sample groups. Here, we provide an overview of technical aspects to look out for in an image cytometry workflow, and discuss issues related to sample preparation, image post-processing and analysis for intra-vital and whole organ imaging.

Idiopathic Generalized Exfoliative Dermatitis and Association with Antihypertensive Drugs and Statins: A Retrospective Case-Control Study.

BACKGROUND/AIMS: We aim to examine the hypothesis that antihypertensive drugs and statins may be responsible for the development of idiopathic generalized exfoliative dermatitis (GED) with a case-control study. METHODS: All inpatients who were hospitalized under the dermatology service at the Tan Tock Seng Hospital, Singapore, between 1 May 2013 and 31 May 2015, were analysed. Idiopathic GED cases had consistent clinical and histological features but no apparent cause despite comprehensive evaluation. Controls were randomly selected from inpatients with other dermatological conditions in a 1: 1 ratio during the same period. Their relationship was analysed using univariate (χ2 or Fisher exact tests) and multivariate logistic regression analysis. RESULTS: There were 78 cases and 83 controls. Of the 78 cases, 42 patients had a history of treatment with antihypertensive drugs or statins. Cases were not found to be more likely on angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, ß-blockers, calcium channel blockers, thiazides or statins compared to controls (OR: 0.81; 95% CI: 0.43-1.51; p = 0.507). CONCLUSIONS: There was insufficient evidence to suggest a significant relationship between the chronic use of antihypertensive drugs or statins and idiopathic GED in this study, despite previous evidence reporting this might be so. Further case-control studies with larger sample sizes are needed to evaluate this association.

Inter- and intra-observer agreement in dermatologists' diagnoses of hyperpigmented facial lesions and development of an algorithm for automated diagnosis.

BACKGROUND: Hyperpigmentation has varied aetio-pathologies. Hence, accurate and reproducible diagnosis of the type of hyperpigmentation is essential for effective management. It is typically made clinically by dermatologists but the rate of inter- and intra-observer agreement/variability is unknown. Hyperpigmented facial lesions are extremely common but access to dermatological services is difficult or costly in most countries. Thus, it is desired to evaluate dermatologists' inter- and intra-observer agreement in the diagnosis and to develop an algorithm for automated diagnosis. MATERIALS AND METHODS: Hyperpigmented lesions on 392 facial images were diagnosed by three experienced dermatologists either jointly or independently, and this process was subsequently repeated for 52 randomly selected images. When there was non-concordance amongst the dermatologists for the diagnosis, a majority decision was taken as correct diagnosis. Inter-observer and intra-observer agreement were analysed for the diagnosis of the hyperpigmented lesions. Thereafter, a multiclass classification method was developed to perform the task in an automatic manner. The developed algorithm was compared and validated against the ground truth derived from the dermatologists. RESULTS: Both inter- and intra-observer agreements are in the moderate range. The algorithm agreed well with the derived ground truth, with a Kappa value of 0.492, which is similar to the Kappa values of inter- and intra- observer agreements. CONCLUSION: The rates of inter- and intra-observer agreement in the diagnosis of hyperpigmented facial lesions amongst dermatologists were moderate in this study. Compared to visual assessment from the dermatologists, automated diagnosis using the developed algorithm achieved a high rate of concordance.

Safety and Antipruritic Efficacy of a Menthol-Containing Moisturizing Cream.

Itch is frequently associated with dermatoses characterized by a defective skin barrier. We formulated an itch-relieving moisturizing cream containing 3% menthol and ceramides. Our aim was to evaluate the safety and antipruritic efficacy of application of this cream in volunteers with and without skin diseases. Volunteers were asked to apply the cream for 1 month on a minimum body surface area of 6%. Safety was assessed by the absence of contact dermatitis or other side effects, using a self-administered questionnaire completed at 5 minutes, 1 week, and 1 month after application. To determine efficacy, volunteers with pruritic dermatoses were asked to grade their average itch intensity at baseline, 1 week, and 1 month after application. Sixty volunteers were recruited, of whom 41 had no skin disease; no adverse events were reported in the latter. Of the 19 volunteers with dermatoses, 18 reportedly had atopic dermatitis. One of the 60 volunteers stopped application due to stinging sensations induced by menthol. Itch scores of volunteers with dermatitis improved from baseline at 1 week (P=.01) and 1 month (P<.01) after application. Application of a 3% menthol-containing moisturizing cream was safe in healthy individuals and participants with dermatitis. In the latter, itch scores were significantly reduced during follow-up.