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IntroductionSingle-cell RNA sequencing (scRNAseq) and flow cytometry studies in skin are methodologically complex and costly, limiting their accessibility to researchers worldwide. Ideally, RNA and protein-based analyses should be performed on the same lesion to obtain more comprehensive data. However, current protocols generally focus on either scRNAseq or flow cytometry of healthy skin.MethodsWe present a novel label-free sample multiplexing strategy, building on the souporcell algorithm, which enables scRNAseq analysis of paired blood and skin samples. Additionally, we provide detailed instructions for simultaneous flow cytometry analysis from the same sample, with necessary adaptations for both healthy and inflamed skin specimens.ResultsThis tissue multiplexing strategy mitigates technical batch effects and reduces costs by 2-4 times compared to existing protocols. We also demonstrate the effects of varying enzymatic incubation durations (1, 3, and 16 hours, with and without enzyme P) on flow cytometry outcomes. Comprehensive explanations of bioinformatic demultiplexing steps and a detailed step-by-step protocol of the entire experimental procedure are included.DiscussionThe protocol outlined in this article will make scRNAseq and flow cytometry analysis of skin samples more accessible to researchers, especially those new to these techniques.

Introduction: Raising the numbers of patch testing in suspected children increases the rate of diagnosis of allergic contact dermatitis and the identification of clinically relevant allergens. Objective: The aim of this study to identify the most frequently observed allergens among Turkish children and adolescents patch tested in 2013-2023. Methods: The patch test results of 160 patients (age ≤18) were analyzed retrospectively. The frequency of contact allergens and distribution of positive results in terms of gender, age group (children and adolescents), and the presence of atopic dermatitis (AD) were identified. Results: Forty-nine patients (30.6%), 34 girls and 15 boys, exhibited positive patch test reactions to a minimum of one allergen, and contact sensitivity was statistically significant higher in girls (P=0.034). The 5 most frequent allergens were nickel sulfate (10.6%), methylchloroisothiazolinone (MCI)/methylisothiazolinone (MI) (8.1%), cobalt chloride (5.6%), p-phenylenediamine (PPD) (5%), and MI (3.5%). No significant association was observed between patch test positivity and age groups (p>.05). Nickel sulfate sensitivity was significantly higher in girls than boys (P=.043). A positive reaction was detected in 31.3% of patients with AD and in 33.7% of those without (P>.05) and a significant relationship was observed between contact sensitivity to fragrance allergens and AD (P=.046). Conclusion: Metals and preservatives represent the most frequent allergens in Turkish children and adolescents. Metal sensitivity is expected to decrease as legislation is enforced. Regulatory measures are now required to reduce MI and MCI/MI contact allergy in Turkey.

Henoch-Schönlein purpura (HSP) is a small vessel vasculitis characterized by the presence of vascular immunoglobulin A deposition that usually presents as non-thrombocytopenic palpable purpura. It primarily affects children and is less common in adults. The incidence of hemorrhagic necrotic skin lesions increases with age, similarly to renal involvement. Warfarin is a widely used oral anticoagulant drug that has rarely been associated with leukocytoclastic vasculitis and allergic interstitial nephritis. We report a patient with HSP who presented with cutaneous ulcerative plaques and proteinuria in the setting of warfarin therapy. We would like to raise the awareness of this potential adverse effect of warfarin for prompt diagnosis.

Abstract Behçet's disease (BD) is a chronic multisystemic inflammatory disease with vasculitis and skin findings. Attacks and remissions characterize the course of the disease. In general, the disease activity decreases with advanced age. Skin symptoms are characteristic of BD and they are the most common findings in the course of the disease. The earliest sign of the disease is almost always recurrent oral aphthous ulcers. Genital ulcers are very specific for BD and they are the equivalent of oral ulcers in the genital area. The most common location is labium majus in females and scrotum in males. The papulopustular lesions are mainly observed on the lower extremities in BD patients. Erythema nodosum‐like lesions and superficial thrombophlebitis are relatively common skin findings. Extragenital ulcerations presenting on the intertriginous area are rare but specific for the disease. The clinical skin manifestations are crucial for the diagnosis of BD, as there is no pathognomonic confirmatory laboratory test.

Introduction: Single-cell RNA sequencing (scRNAseq) and flow cytometry studies in skin are methodologically complex and costly, limiting their accessibility to researchers worldwide. Ideally, RNA and protein-based analyses should be performed on the same lesion to obtain more comprehensive data. However, current protocols generally focus on either scRNAseq or flow cytometry of healthy skin. Methods: We present a novel label-free sample multiplexing strategy, building on the souporcell algorithm, which enables scRNAseq analysis of paired blood and skin samples. Additionally, we provide detailed instructions for simultaneous flow cytometry analysis from the same sample, with necessary adaptations for both healthy and inflamed skin specimens. Results: This tissue multiplexing strategy mitigates technical batch effects and reduces costs by 2-4 times compared to existing protocols. We also demonstrate the effects of varying enzymatic incubation durations (1, 3, and 16 hours, with and without enzyme P) on flow cytometry outcomes. Comprehensive explanations of bioinformatic demultiplexing steps and a detailed step-by-step protocol of the entire experimental procedure are included. Discussion: The protocol outlined in this article will make scRNAseq and flow cytometry analysis of skin samples more accessible to researchers, especially those new to these techniques. [ABSTRACT FROM AUTHOR]

Summary: Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells (PBMCs). We describe the steps for isolating target cells and their in-column surface biotinylation, followed by isolation and mass spectrometry analysis of biotinylated proteins. The protocol enables in-column surface biotinylation of specific cell subsets with minimal membrane disruption. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

29-year-old woman presented with dark-colored raised lesions on both eyelids since early childhood. Ophthalmological examination revealed pigmented verrucous lesions on her upper and lower eyelids bilaterally. The patient had a history of generalized tonic-clonic seizures. Dermatological examination revealed hyperpigmented verrucous plaques arranged along lines of Blaschko on the neck, trunk, and arms. On the basis of these findings, the diagnosis of epidermal nevus syndrome (ENS) was made. She had surgery for debulking of the lesions. Histological analysis revealed hyperkeratosis with foci of parakeratosis, acanthosis, and papillomatosis, consistent with linear verrucous epidermal nevus. Postoperative residual lesions did not respond to oral acitretin therapy (10 mg/kg/day for 2 months). Systematized ENS can rarely cause linear verrucous nevi on the upper and lower eyelids on both sides. These patients should be investigated for accompanying systemic anomalies and followed for potential malignant transformation of the skin lesions.

Abstract Small‐scale wireless magnetic robots and devices offer an effective solution to operations in hard‐to‐reach and high‐risk enclosed places, such as inside the human body, nuclear plants, and vehicle infrastructure. In order to obtain functionalities beyond the capability of magnetic forces and torques exerted on magnetic materials used in these robotic devices, electronics need to be also integrated into them. However, their capabilities and power sources are still very limited compared to their larger‐scale counterparts due to their much smaller sizes. Here, groups of milli/centimeter‐scale wireless magnetic modules are shown to enable on‐site electronic circuit construction and operation of highly demanding wireless electrical devices with no batteries, that is, with wireless power. Moreover, the mobility of the modular components brings remote modification and reconfiguration capabilities. When these small‐scale robotic modules are remotely assembled into specific geometries, they can achieve, if not impossible, challenging electrical tasks for individual modules. Using such a method, several wireless and battery‐free robotic devices are demonstrated using milli/centimeter‐scale robotic modules, such as a wireless circuit to power light‐emitting diodes with lower external fields, a device to actuate relatively high force‐output shape memory alloy actuators, and a wireless force sensor, all of which can be modified on‐site.

Syntaxin-1 (STX1) and Munc18-1 are two requisite components of synaptic vesicular release machinery, so much so synaptic transmission cannot proceed in their absence. They form a tight complex through two major binding modes: through STX1’s N-peptide and through STX1’s closed conformation driven by its Habc- domain. However, physiological roles of these two reportedly different binding modes in synapses are still controversial. Here we characterized the roles of STX1’s N-peptide, Habc-domain, and open conformation with and without N-peptide deletion using our STX1-null mouse model system and exogenous reintroduction of STX1A mutants. We show, on the contrary to the general view, that the Habc-domain is absolutely required and N-peptide is dispensable for synaptic transmission. However, STX1A’s N-peptide plays a regulatory role, particularly in the Ca2+-sensitivity and the short-term plasticity of vesicular release, whereas STX1’s open conformation governs the vesicle fusogenicity. Strikingly, we also show neurotransmitter release still proceeds when the two interaction modes between STX1A and Munc18-1 are presumably intervened, necessitating a refinement of the conceptualization of STX1A–Munc18-1 interaction.

Abstract Magnetic resonance imaging (MRI) scanners do not provide only high‐resolution medical imaging but also magnetic robot actuation and tracking. However, the rotational motion capabilities of MRI‐powered wireless magnetic capsule‐type robots have been limited due to the very high axial magnetic field inside the MRI scanner. Medical functionalities of such robots also remain a challenge due to the miniature robot designs. Therefore, a wireless capsule‐type reversible orientation‐locking robot (REVOLBOT) is proposed that has decoupled translational motion and planar orientation change capability by locking and unlocking the rotation of a spherical ferrous bead inside the robot on demand. Such an on‐demand locking/unlocking mechanism is achieved by a phase‐changing wax material in which the ferrous bead is embedded inside. Controlled and on‐demand hyperthermia and drug delivery using wireless power transfer‐based Joule heating induced by external alternating magnetic fields are the additional features of this robot. The experimental feasibility of the REVOLBOT prototype with steerable navigation, medical function, and MRI tracking capabilities with an 1.33 Hz scan rate is demonstrated inside a preclinical 7T small‐animal MRI scanner. The proposed robot has the potential for future clinical use in teleoperated minimally invasive treatment procedures with hyperthermia and drug delivery capabilities while being wirelessly powered and monitored inside MRI scanners.

Magnetic resonance imaging (MRI) system–driven medical robotics is an emerging field that aims to use clinical MRI systems not only for medical imaging but also for actuation, localization, and control of medical robots. Submillimeter scale resolution of MR images for soft tissues combined with the electromagnetic gradient coil–based magnetic actuation available inside MR scanners can enable theranostic applications of medical robots for precise image‐guided minimally invasive interventions. MRI‐driven robotics typically does not introduce new MRI instrumentation for actuation but instead focuses on converting already available instrumentation for robotic purposes. To use the advantages of this technology, various medical devices such as untethered mobile magnetic robots and tethered active catheters have been designed to be powered magnetically inside MRI systems. Herein, the state‐of‐the‐art progress, challenges, and future directions of MRI‐driven medical robotic systems are reviewed.

Background: Preservatives are a frequent cause of allergic contact dermatitis (ACD) and have caused numerous epidemics. Objectives: The objective of this study is to determine the prevalence of preservative sensitivity, assess the change in the frequency of sensitivity, identify new preservatives with increased sensitivity rates, and evaluate the situation in Turkey by comparing our findings with current literature. Methods: A total of 201 patients diagnosed with ACD between 2018 and 2020, were patch tested with the European baseline series and additional seven preservative haptens. The change in the prevalence of sensitivity to each preservative hapten was investigated by comparing the data from the study conducted in our department between 2000 and 2004. Results: Results showed that 17.4% (n = 35) of the patients were positive to preservatives. Comparison with previous data from 2000 to 2004 revealed an increase in the frequency of sensitization. The most prevalent allergen was methyldibromo glutaronitrile (9.5%), followed by methylchloroisothiazolinone/methylisothiazolinone (6.5%), and methylisothiazolinone (5%). Conclusion: The increase in preservative sensitivity in Turkey is the most remarkable finding. Although MDBGN was prohibited in cosmetic products, MCI/MI and MI are still widely used. Our findings suggest that awareness of preservative sensitivity should be increased and additional precautions should be taken, also in Turkey, regarding the use of preservatives. [ABSTRACT FROM AUTHOR]