CRISPR-Cas9 editing of the arginine-vasopressin V1a receptor produces paradoxical changes in social behavior in Syrian hamsters.

Studies from a variety of species indicate that arginine­vasopressin (AVP) and its V1a receptor (Avpr1a) play a critical role in the regulation of a range of social behaviors by their actions in the social behavior neural network. To further investigate the role of AVPRs in social behavior, we performed CRISPR-Cas9­mediated editing at the Avpr1a gene via pronuclear microinjections in Syrian hamsters (Mesocricetus auratus), a species used extensively in behavioral neuroendocrinology because they produce a rich suite of social behaviors. Using this germ-line gene-editing approach, we generated a stable line of hamsters with a frame-shift mutation in the Avpr1a gene resulting in the null expression of functional Avpr1as. Avpr1a knockout (KO) hamsters exhibited a complete lack of Avpr1a-specific autoradiographic binding throughout the brain, behavioral insensitivity to centrally administered AVP, and no pressor response to a peripherally injected Avpr1a-specific agonist, thus confirming the absence of functional Avpr1as in the brain and periphery. Contradictory to expectations, Avpr1a KO hamsters exhibited substantially higher levels of conspecific social communication (i.e., odor-stimulated flank marking) than their wild-type (WT) littermates. Furthermore, sex differences in aggression were absent, as both male and female KOs exhibited more aggression toward same-sex conspecifics than did their WT littermates. Taken together, these data emphasize the importance of comparative studies employing gene-editing approaches and suggest the startling possibility that Avpr1a-specific modulation of the social behavior neural network may be more inhibitory than permissive.

Clinical grade multiparametric cell sorting and gene-marking of regulatory T cells.

BACKGROUND AIMS: Regulatory T cells (Tregs) are the main mediators of peripheral tolerance. Treg-directed therapy has shown promising results in preclinical studies of diverse immunopathologies. At present, the clinical applicability of adoptive Treg transfer is limited by difficulties in generating Tregs at sufficient cell dose and purity. METHODS: We developed a Good Manufacturing Practice (GMP) compliant method based on closed-system multiparametric Fluorescence-Activated Cell Sorting (FACS) to purify Tregs, which are then expanded in vitro and gene-marked with a clinical grade retroviral vector to enable in vivo fate tracking. Following small-scale optimization, we conducted four clinical-scale processing runs. RESULTS: We showed that Tregs could be enriched to 87- 92% purity following FACS-sorting, and expanded and transduced to yield clinically relevant cell dose of 136-732×106 gene-marked cells, sufficient for a cell dose of at least 2 × 106 cells/kg. The expanded Tregs were highly demethylated in the FOXP3 Treg-specific demethylated region (TSDR), consistent with bona fide natural Tregs. They were suppressive in vitro, but a small percentage could secrete proinflammatory cytokines, including interferon-γ and interleukin-17A. CONCLUSIONS: This study demonstrated the feasibility of isolating, expanding and gene-marking Tregs in clinical scale, thus paving the way for future phase I trials that will advance knowledge about the in vivo fate of transferred Tregs and its relationship with concomitant Treg-directed pharmacotherapy and clinical response.

Male dingo urinary scents code for age class and wild dingoes respond to this information.

Chemical information in canid urine has been implicated in territoriality and influences the spacing of individuals. We identified the key volatile organic compound (VOC) components in dingo (Canis lupus dingo) urine and investigated the potential role of scents in territorial spacing. VOC analysis, using headspace gas chromatography-mass spectrometry (GC-MS), demonstrated that the information in fresh urine from adult male dingoes was sufficient to allow statistical classification into age categories. Discriminant function analyses demonstrated that the relative amounts or combinations of key VOCs from pre-prime (3-4 years), prime (5-9 years), and post-prime (≥10 years) males varied between these age categories, and that scents exposed to the environment for 4 (but not 33) days could still be classified to age categories. Further, a field experiment showed that dingoes spent less time in the vicinity of prime male dingo scents than other scents. Collectively, these results indicate that age-related scent differences may be discriminable by dingoes. Previous authors have suggested the potential to use scent as a management tool for wild canids by creating an artificial territorial boundary/barrier. Our results suggest that identifying the specific signals in prime-age male scents could facilitate the development of scent-based tools for non-lethal management.

Effects of intrauterine position during gestation on specific endocrine and behavioral parameters that impact reproduction in domestic rabbits.

Prior studies from others, performed in a different breed, reported that doe rabbits developing between two male siblings (2 M) during gestation display characteristics indicative of masculinization: larger anogenital distance (AGD), larger submandibular glands, and higher chinning frequency than females with zero (0 M) or one (1 M) contiguous brothers. Similar effects are provoked by injecting androgens to the pregnant doe suggesting that prenatal androgen exposure masculinizes female embryos. To further understand the scope of such masculinization we compared 0 M, 1 M, and 2 M females regarding behavioral, neuroendocrine, and somatic parameters, related or not to reproduction. IUP did not impact: body weight, sexual receptivity, mating-induced LH secretion, maternal nest-building, litter size, or milk output. At puberty: a) chinning frequency was: 0 M and males>1 M and 2 M; b) ambulation in open field was lowest in 1 M females and males. IUP effects on AGD were significant only on postnatal day 1: 0 M, 1 M, and males>2 M, in contrast to earlier study. Willingness to nurse at delivery was less frequent in 2 M than in 1 M and 0 M does and correlated with nursing occurrence across lactation. Does that did not nurse at parturition delivered fewer kits/min than those that nursed then, regardless of IUP. The duration of nursing bouts across lactation was significantly longer in the1 M and 2 M does that showed this behavior on postpartum days 1-20. Our findings indicate that IUP is associated with alterations in specific aspects of postpartum maternal behavior.

Diagnostic Performance of Near-Infrared Fluorescent Marking Clips in Laparoscopic Gastrectomy.

INTRODUCTION: Accurate tumor localization and resection margin acquisition are essential in gastric cancer surgery. Preoperative placement of marking clips in laparoscopic gastrectomy as well as intraoperative gastroscopy can be used for gastric cancer surgery. However, these procedures are not available at all institutions. We conducted a prospective clinical trial to investigate the diagnostic performance of near-infrared fluorescent clips (ZEOCLIP FS) in laparoscopic gastrectomy. MATERIALS AND METHODS: Patients with gastric cancer or neuroendocrine tumor in whom laparoscopic distal, pylorus-preserving, or proximal gastrectomy was planned were enrolled (n = 20) in this study. Fluorescent clips were placed proximal and/or distal to the tumor via gastroscopy on the day before surgery. During surgery, the clips were detected using a fluorescent laparoscope, and suturing was performed where fluorescence was detected. The clip locations were then confirmed via gastroscopy, and the stomach was transected. The primary endpoint was the detection rate of the marking clips using fluorescence, and the secondary endpoints were complications and distance between the clips and stitches. RESULTS: Among the 20 patients enrolled, distal and pylorus-preserving gastrectomies were performed in 18 and 2 patients, respectively. All clips were detected in 15 patients, indicating a detection rate of 75.0% (90% confidence interval: 54.4%-89.6%). Furthermore, no complications related to the clips were observed. The median distance between the clips and stitches was 5 (range, 0-10) mm. CONCLUSIONS: We report the feasibility and safety of preoperative placement and intraoperative detection of near-infrared fluorescent marking clips in laparoscopic gastrectomy.

Competition through ritualized aggressive interactions between sympatric colonies in solitary foraging neotropical ants.

Understanding the structure of food competition between conspecifics in their natural settings is paramount to addressing more complex questions in ecology, evolution, and conservation. While much research on ants focuses on aggressive food competition between large and foraging trail-using societies, we lack a thorough understanding of inter-colony competition in socially less derived, solitarily foraging species. To fill this gap, we explored the activity of ten neighbouring colonies of the giant ant Dinoponera quadriceps, monitoring 2513 foraging trips of hundreds of workers and all its inter-individual interactions. We found that, on encountering, workers from different colonies rarely engaged in aggressive fights but instead avoided each other or performed ritualised agonistic bouts. We discovered that during foraging trips, a few workers within each colony repeatedly rubbed their gaster on the substrate, a behaviour not observed in the field before. We propose that workers use this behaviour to mark the foraging area and mark more frequently in its periphery. Only 25% of the individuals specialised in this behaviour, and we hypothesise that the specialisation results from the history of interactions and experience of individual foragers. Our study suggests that workers of contiguous D. quadriceps colonies engage in low-risk conflict, mainly displaying ritualised behaviours. As these small societies mainly rely on tiny, unpredictably scattered, albeit abundant in the environment, arthropod prey, and not on persistent food sources, they do not aggressively defend exclusive foraging territories. On the other hand, colonies rely on large overlapping foraging areas to sustain their survival and growth, most often tolerating foragers from nearby colonies. We discuss whether this type of competitive interaction is expected in all solitary foraging species.

Gradually increasing context-sensitivity shapes the development of children's verb marking: A corpus study.

There is substantial evidence that children's apparent omission of grammatical morphemes in utterances such as "She play tennis" and "Mummy eating" is in fact errors of commission in which contextually licensed unmarked forms encountered in the input are reproduced in a context-blind fashion. So how do children stop making such errors? In this study, we test the assumption that children's ability to recover from error is related to their developing sensitivity to longer-range dependencies. We use a pre-registered corpus analysis to explore the predictive value of different cues with regards to children's verb-marking errors and observe a developmental pattern consistent with this account. We look at context-independent cues (the identity of the specific verb being used) and at the relative value of context-dependent cues (the identity of the specific subject+verb sequence being used). We find that the only consistent effect across a group of 2- to 3-year-olds and a group of 3- to 4-year-olds is the relative frequency of unmarked forms of specific subject+verb sequences being used. The relative frequency of unmarked forms of the verb alone is predictive only in the younger age group. This is consistent with an account in which children recover from making errors by becoming progressively more sensitive to context, at first the immediately preceding lexical contexts (e.g., the subject that precedes the verb) and eventually more distant grammatical markers (e.g., the fronted auxiliary that precedes the subject in questions). RESEARCH HIGHLIGHTS: We provide a corpus analysis investigating input effects on young children's verb-marking errors (e.g., Mummy go) across development (between 2 and 4 years of age). We find evidence that these apparent errors of omission are in fact input-driven errors of commission that persist into the third year of life. We compare the relative effect on error rates of context-independent (e.g., verb) and context-dependent (e.g., subject+verb sequence) cues across developmental time. Our findings support the proposal that children recover from making verb-marking errors by becoming progressively more sensitive to preceding context.

Mesocarnivores vary in their spatiotemporal avoidance strategies at communications hubs of an apex carnivore.

Mesocarnivores face interspecific competition and risk intraguild predation when sharing resources with apex carnivores. Within a landscape, carnivores across trophic levels may use the same communication hubs, which provide a mix of risks (injury/death) and rewards (gaining information) for subordinate species. We predicted that mesocarnivores would employ different strategies to avoid apex carnivores at shared communication hubs, depending on their trophic position. To test our prediction, we examined how different subordinate carnivore species in the Santa Cruz Mountains of California, USA, manage spatial overlap with pumas (Puma concolor), both at communication hubs and across a landscape-level camera trap array. We estimated species-specific occurrence, visitation rates, temporal overlap, and Avoidance-Attraction Ratios from camera traps and tested for differences between the two types of sites. We found that mesocarnivores generally avoided pumas at communication hubs, and this became more pronounced when pumas scent-marked during their most recent visit. Coyotes (Canis latrans), the pumas' closest subordinate competitor in our system, exhibited the strongest avoidance at communication hubs. Gray foxes (Urocyon cinereoargenteus) avoided pumas the least, which may suggest possible benefits from pumas suppressing coyotes. Overall, mesocarnivores exhibited various spatiotemporal avoidance strategies at communication hubs rather than outright avoidance, likely because they benefit from information gained while 'eavesdropping' on puma activity. Variability in avoidance strategies may be due to differential predation risks, as apex carnivores often interact more aggressively with their closest competitors. Combined, our results show how apex carnivores trigger complex species interactions across the entire carnivore guild and how trophic position determines behavioral responses and subsequent space use of subordinate mesocarnivores across the landscape.

Another view of sequential sampling in the birth process with immigration.

We explore properties of the family sizes arising in a linear birth process with immigration (BI). In particular, we study the correlation of the number of families observed during consecutive disjoint intervals of time. Letting S(a, b) be the number of families observed in (a, b), we study the expected sample variance and its asymptotics for p consecutive sequential samples [Formula: see text], for [Formula: see text]. By conditioning on the sizes of the samples, we provide a connection between [Formula: see text] and p sequential samples of sizes [Formula: see text], drawn from a single run of a Chinese Restaurant Process. Properties of the latter were studied in da Silva et al. (Bernoulli 29:1166-1194, 2023. https://doi.org/10.3150/22-BEJ1494 ). We show how the continuous-time framework helps to make asymptotic calculations easier than its discrete-time counterpart. As an application, for a specific choice of [Formula: see text], where the lengths of intervals are logarithmically equal, we revisit Fisher's 1943 multi-sampling problem and give another explanation of what Fisher's model could have meant in the world of sequential samples drawn from a BI process.

The effect of the attitude towards risk/ambiguity on examination grades: cross-sectional study in a Portuguese medical school.

Given the high prevalence of multiple-choice examinations with formula scoring in medical training, several studies have tried to identify other factors in addition to the degree of knowledge of students which influence their response patterns. This study aims to measure the effect of students' attitude towards risk and ambiguity on their number of correct, wrong, and blank answers. In October 2018, 233 3rd year medical students from the Faculty of Medicine of the University of Porto, in Porto, Portugal, completed a questionnaire which assessed the student's attitudes towards risk and ambiguity, and aversion to ambiguity in medicine. Simple and multiple regression models and the respective regression coefficients were used to measure the association between the students' attitudes, and their answers in two examinations that they had taken in June 2018. Having an intermediate level of ambiguity aversion in medicine (as opposed to a very high or low level) was associated with a significant increase in the number of correct answers and decrease in the number of blank answers in the first examination. In the second examination, high levels of ambiguity aversion in medicine were associated with a decrease in the number of wrong answers. Attitude towards risk, tolerance for ambiguity, and gender did not show significant association with the number of correct, wrong, and blank answers for either examination. Students' ambiguity aversion in medicine is correlated with their performance in multiple-choice examinations with negative marking. Therefore, it is suggested the planning and implementation of counselling sessions with medical students regarding the possible impact of ambiguity aversion on their performance in multiple-choice questions with negative marking.

Conspicuity of staircase configuration: Effects of markings and contrast.

PURPOSE: To be able to walk safely up or down a staircase, we must be able to judge the configuration and slope of the staircase and our viewing position. Adding markings to the stairs might help form correct perceptions of the staircase geometry. In this study, we examined how visual judgements about staircase configuration are affected by different marking patterns. METHODS: Fifteen normally sighted young participants viewed computer-generated images of staircases as seen from the top landing of the stairs. Marking patterns included contrasting baseboard, transverse edge-stripes, longitudinal side-stripes, longitudinal stripes, diamond patterns, longitudinal stripes extended to landing and diamond patterns extended to landing. For comparison, we included the no-marking condition as a control. We tested several contrast levels of marking patterns (3.2%-50%), pitch lines of the staircases (shallow/medium/steep) and viewing positions (left/centre/right). The effect of the overall shape cue of the staircase on participants' performance was also evaluated. We measured participants' accuracies in judging whether the staircase was shallow, medium or steep, and whether the viewing position was located to the left, centre or right. RESULTS: Transverse edge-stripes markings yielded fewer underestimations of slope (9% [transverse] vs. 18% [others]) when compared with other markers. The presence of an overall shape cue helped both slope (67% [presence] vs. 51% [absence]) and viewing position judgements (79% [presence] vs. 62% [absence]). When the overall shape cue was present, only the transverse edge-stripes markings yielded a significant improvement in performance (compared with no-marking condition). When the cue was absent, performance was significantly better with markings with high and moderate contrasts. CONCLUSIONS: Adding marking patterns such as high-contrast transverse stripes to stairs may help enhance the visibility of the stairs and judgements of staircase geometry. This might be particularly useful for people with visual impairment or normally sighted individuals under compromised environmental conditions.

The social and defensive function of olfactory behaviors in a pair-living sexually monogamous primate.

Olfactory behaviors serve a wide variety of social functions in mammals. Odor may signal information about attributes of individuals important for mating and reproduction. Olfactory behaviors, such as scent-marking, may also function as part of home range or resource defense strategies. We assessed the potential social and home range defense functions of olfactory behavior in a pair-living and sexually monogamous primate, Azara's owl monkey (Aotus azarae), in the Argentinian Chaco. This is the most extensive investigation of owl monkey olfactory behaviors in the wild. Individuals regularly performed olfactory behaviors (group mean + SD = 1.3 + 0.5 per hour). The patterns were generally comparable to those observed in studies of captive owl monkeys, except that urine washing was the most common behavior in the wild, as opposed to scent-marking and genital inspections. Most olfactory behaviors were performed by adults, and there were striking sex differences in genital inspections: almost all consisted of an adult male inspecting the paired adult female. These findings suggest that olfactory behaviors play an important role in signaling and coordinating reproduction among owl monkeys, particularly during periods of female conception and pregnancy. Additionally, our research indicates that these behaviors may also serve as a defense strategy for maintaining the core area of their home ranges. This study offers the first assessment of the role of olfactory behaviors in reproductive contexts and home range defense in pair-living, monogamous platyrrhine primates.

Standard methods for marking caudate amphibians do not impair animal welfare over the short term: An experimental approach.

Major advancements in ecology and biodiversity conservation have been made thanks to methods for marking and individually tracking animals. Marking animals is both widely used and controversial due to the potential consequences for animal welfare, which are often incompletely evaluated prior to implementation. Two outstanding knowledge gaps concerning the welfare consequences of individual marking are their short-term behavioural impacts and the relative impacts from marking versus the handling of animals while carrying out procedures. We addressed these knowledge gaps through an experimental study of alpine newts (Ichthyosaura alpestris) in which we varied handling and marking procedures. Examining individual responses to handling, toe clipping and visible implant elastomer (VIE) injection over 21 days showed that handling and marking elicited increased newt activity and hesitancy to feed compared to animals that did not get handled or marked. These effects were apparent even when animals were handled only (not marked), and marking did not further increase the magnitude of responses. Increases in newt activity and feeding hesitancy were transient; they were not observed in the weeks following handling and marking. While previous studies emphasise the welfare impacts of marking procedures themselves, these findings highlight that handling alone can elicit behavioural changes with possible costs to welfare. Yet, the transient nature of behavioural responses suggests that immediate costs of handling may be subsequently compensated for in the short term.

Twelve tips for developing effective marking schemes for constructed-response examination questions.

Constructed-response questions (CRQs) require effective marking schemes to ensure that the intended learning objectives and/or professional competencies are appropriately addressed, and valid inferences regarding examinee competence are drawn from such assessments. While the educational literature on writing rubrics has proliferated in recent years, this is largely targeted at classroom use and formative purposes. There is comparatively little guidance on how to develop appropriate marking schemes for summative assessment contexts. The different purposes mean that different principles and practices apply to mark schemes for examinations. In this article, we draw on the educational literature as well as our own practical experience of working with medical and health professional educators on their questions and marking schemes to offer 12 key principles or tips for designing and implementing effective marking schemes.

Superior Medial-Based Breast Reduction with Inverted T Incision: A Precise Description of Our Refined Preoperative Marking and Operative Technique.

The purpose of this paper was to compile a thoroughly elaborated step-by-step guide for the preoperative marking and operative technique for superior medial pedicle inverted T breast reduction based on our long experience and technical refinements. 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 .

Optimizing the mass marking of fish with alizarin red S: an example with glass eels.

Fish marking is an essential tool for fisheries management, especially for evaluating the stocking of endangered fish species to support conservation and sustainable use of fish stocks. Batch marking of young European eels Anguilla anguilla (L.) prior to stocking is recommended as the benefits of stocking for the spawning stock can be evaluated by recapturing marked fish over time, therefore mass marking of young eels with substances such as alizarin red S (ARS) is becoming increasingly important. To improve the marking method and reduce marking costs when immersing glass eels in an ARS solution, eight laboratory experiments under varying conditions (e.g., temperature, ARS concentration, immersion time, osmotic induction, fish density) and with ARS from different suppliers were carried out. The results show that optimal marking of glass eels can be carried out in the field or during transport by putting approximately 50 g of glass eels per liter in 150 mg L-1 ARS solution for 3 h at 10-15°C. Lower concentrations did not result in reliable marking. Water temperatures of 5°C and below can have a stunning effect on the eels and increase mortality significantly, regardless of the concentration of ARS. Glass eel densities below 50 g L-1 in the marking bath increase marking costs unnecessarily, while a higher density of 100 g L-1 resulted in significantly higher mortality and lower marking success. A somewhat more difficult but less expensive alternative is to bathe the fish in a saline solution of 1% (10 PSU) of 80 mg L-1 ARS for 3 h at 10°C. Costs can also be significantly reduced by choice of supplier for ARS, but care should be taken as the quality of the powder appears to vary (mean percentage of sufficiently marked eels ranged from 59% to 91% among suppliers in the present study) and can lead to marking failure. The optimal marking conditions can help ensure that stocked glass eels can be reliably identified in future studies to assess stocking benefits while reducing costs.

The GoldX Fiducial Eraser.

Gold nanoparticles with sizes in the range of 5-15 nm are a standard method of providing fiducial markers to assist with alignment during reconstruction in cryogenic electron tomography. However, due to their high electron density and resulting contrast when compared to standard cellular or biological samples, they introduce artifacts such as streaking in the reconstructed tomograms. Here, we demonstrate a tool that automatically detects these nanoparticles and suppresses them by replacing them with a local background as a post-processing step, providing a cleaner tomogram without removing any sample relevant information or introducing new artifacts or edge effects from uniform density replacements.

I spy: Factors influencing the observation of oxytetracycline in calcified structures of fishes viewed using standard light and fluorescence microscopy.

The antibiotic oxytetracycline (OTC) is a fluorochrome marker, and fluorescence microscopy is used to view OTC marks in fishes' calcified structures. However, OTC marks have been observed in calcified structures using standard light microscopy for multiple species. Therefore, we conducted an experiment to investigate potential factors (i.e., season, total length of fish, growth rate, and sex) influencing the observation of OTC in calcified structures (otoliths and fin rays or spines) from channel catfish Ictalurus punctatus, gray redhorse Moxostoma congestum, Guadalupe bass Mircopterus treculii, and redbreast sunfish Lepomis auritus viewed using standard light and fluorescence microscopy. OTC stains were not observed in any otoliths under standard light; however, OTC marks were commonly observed in I. punctatus spines using standard light microscopy (56.2%). Ninety-nine percent of otoliths and 88.9% of spines and fin rays had a visible fluorescent OTC mark when viewed using fluorescence microscopy. There was a negative relationship between the observed OTC mark and total length of fish for each season, but fish injected in the summer had the most structures with an observed OTC mark under either light condition. Understanding how OTC marking is affected by biological processes and environmental conditions will assist in future studies that rely on chemical marking of calcified structures by increasing efficacy of OTC marking and interpretation of marks.

Donning and doffing of personal protective equipment for health care workers in a tertiary hospital in China: A simulation study.

The application and removal of personal protective equipment (PPE) by health care workers (HCWs) is pivotal to their health and safety and the comprehensive efficacy of hospital infection control measures. This investigation was orchestrated to elucidate the challenges that HCWs may encounter during the donning and doffing of PPE. A total of 110 participants from a tertiary hospital in China were engaged. The study employed fluorescent markers to mimic the exposure of HCWs to tainted body fluids, quantified the contamination outcomes, and evaluated adherence to procedures for donning and doffing. Factors including gender, educational background, and the timing of the most recent instruction on PPE donning and doffing were found to influence the occurrence of contamination points (p < 0.05). No significant differences were identified in contamination frequency when assessing age, body mass index (BMI), occupation, educational background, positional title, working tenure, and experience in managing respiratory infectious diseases (p > 0.05). Predominant contamination sites for fluorescent marker residue included the shoulder (32.73%), neck (21.82%), forearm (16.36%), chest (12.73%), and abdomen (11.82%), with the shoulder being the most contaminated body part. A majority of HCWs exhibited susceptibility to errors during the removal of protective clothing, boot covers, and gloves. The contamination frequency was observed to be correlated with the timing of the last PPE training, educational background, and gender. In acknowledging the intricacy of PPE removal and the deficiencies in HCWs' removal techniques, there emerges a perpetual necessity to refine training methodologies and perpetuate regular PPE instruction.

Higher intraoperative detection rate of suspicious non-palpable lesions using wire marking in skin cancer patients.

BACKGROUND: Follow-up protocols in patients after complete resection of high-risk cutaneous tumors lead to a discovery of metastases in very early stages, but surgery on non-palpable lesions proves to be challenging. PATIENTS AND METHODS: In this monocenter retrospective study 39 patients suffering from malignant skin tumors with suspicious non-palpable lesions located in the lymph nodes (90%) or deep subcutaneously/intramuscularly (10%) were included. In 21 patients the lesions were excised under ultrasound guidance, and 18 patients received a wire marking before surgery. Both patient groups were compared regarding successful intraoperative finding of the lesion, duration of the procedure, and complications. RESULTS: Wire marking led to a significantly higher intraoperative detection rate of 100% versus 76% (p < 0.05). The average time needed for the complete procedure (p = 0.91) or the rate of complications (p = 0.70) did not differ significantly between both groups. The size of the malignant lesions successfully removed by wire marking was significantly smaller (p < 0.05). Of all 34 detected lesions only 20 (58.8%) were confirmed to be malignant. CONCLUSIONS: Wire marking increases the detection rate of non-palpable suspicious subcutaneous or lymphatic lesions. It leads to earlier diagnosis of metastasis but also allows to avoid unnecessary complete lymph node dissection.