Dissection, Histological Processing, and Gene Expression Analysis of Murine Supraclavicular Brown Adipose Tissue.

Brown adipose tissue (BAT)-mediated thermogenesis plays an important role in the regulation of metabolism, and its morphology and function can be greatly impacted by environmental stimuli in mice and humans. Currently, murine interscapular BAT (iBAT), which is located between two scapulae in the upper dorsal flank of mice, is the main BAT depot used by research laboratories to study BAT function. Recently, a few previously unknown BAT depots were identified in mice, including one analogous to human supraclavicular brown adipose tissue. Unlike iBAT, murine supraclavicular brown adipose tissue (scBAT) is situated in the intermediate layer of the neck and thus cannot be accessed as readily. To facilitate the study of newly identified mouse scBAT, presented herein is a protocol detailing the steps to dissect intact scBAT from postnatal and adult mice. Due to scBAT's small size relative to other adipose depots, procedures have been modified and optimized specifically for processing scBAT. Among these modifications is the use of a dissecting microscope during tissue collection to increase the precision and homogenization of frozen scBAT samples to raise the efficiency of subsequent qPCR analysis. With these optimizations, the identification of, morphological appearance of, and molecular characterization of the scBAT can be determined in mice.

A realistic aneurysm clipping simulation combining 3D-printed and placenta-based models-how I do it.

INTRODUCTION: Neurovascular surgery, particularly aneurysm clipping, is a critical skill for aspiring neurosurgeons. However, hands-on training opportunities are limited, especially with the growing popularity of endovascular techniques. To address this challenge, we present a novel neurovascular surgical training station that combines synthetic 3D-printed models with placental vascular structures to create a semi-realistic surgical field. METHODS: Our model consists of three components: a 3D-printed skull replica with anatomical landmarks, a malleable silicone parenchyma with a Sylvian fissure, and vascular layers (placenta). The placental vascular layer is catheterized and perfused to replicate pulsatile flow, offering a realistic aneurysm simulation. This innovative training station provides a cost-effective solution (approximately 200 USD once) without ethical constraints. Surgeons can practice essential skills such as Sylvian fissure dissection, managing anatomical constraints like bone, and achieving proximal vascular control. The model's realism allows for training in various scenarios, including clipping with different hand orientations and handling ruptures realistically. CONCLUSION: Our neurovascular surgical station bridges the gap between existing training models, offering affordability, ecological considerations, and minimal ethical concerns. It empowers neurosurgery residents to refine their skills in handling both emergencies and elective cases under close-to-real surgical conditions, with the potential for independent practice and senior supervision.

Quantification of surgical workflow during robotic proctectomy.

BACKGROUND: Surgical workflow assessments offer insight regarding procedure variability. We utilised an objective method to evaluate workflow during robotic proctectomy (RP). METHODS: We annotated 31 RPs and used Spearman's correlation to measure the correlation of step time and step visit frequency with console time (CT) and total operative time (TOT). RESULTS: Strong correlations were seen with CT and step times for inferior mesenteric vein dissection and ligation (ρ = 0.60, ρ = 0.60), lateral-to-medial splenic flexure mobilisation (SFM) (ρ = 0.63), left rectal dissection (ρ = 0.64) and mesorectal division (ρ = 0.71). CT correlated strongly with medial-to-lateral (ρ = 0.75) and supracolic SFM visit frequency (ρ = 0.65). TOT correlated strongly with initial exposure time (ρ = 0.60), and medial-to-lateral (ρ = 0.67) and supracolic SFM visit frequency (ρ = 0.65). CONCLUSION: This study correlates surgical steps with CT and TOT through standardised annotation, providing an objective approach to quantify workflow.

Cloning and functional analysis of ZmMADS42 gene in maize.

Maize (Zea mays L.) is the most important cereal crop in the world. Flowering period and photoperiod play important roles in the reproductive development of maize. This study, investigated ZmMADS42, a gene that is highly expressed in the shoot apical meristem. Agrobacterium infection was used to successfully obtain overexpressed ZmMADS42 plants. Fluorescence quantitative PCR revealed that the expression of the ZmMADS42 gene in the shoot apical meristem of transgenic plants was 2.8 times higher than that of the wild-type(WT). In addition, the expression of the ZmMADS42 gene in the endosperm was 2.4 times higher than that in the wild-type. The seed width of the T2 generation increased by 5.35%, whereas the seed length decreased by 7.78% compared with that of the wild-type. Dissection of the shoot tips of transgenic and wild-type plants from the 7-leaf stage to the 9-leaf stage revealed that the transgenic plants entered the differentiation stage earlier and exhibited more tassel meristems during their vegetative growth period. The mature transgenic plants were approximately 20 cm shorter in height and had a lower panicle position than the wild-type plants. Comparing the flowering period, the tasseling, powdering, and silking stages of the transgenic plants occurred 10 days earlier than those of the wild-type plants. The results showed that the ZmMADS42 gene played a significant role in regulating the flowering period and plant height of maize.

A national survey to assess the state of anatomy teaching in France across various disciplines and professions.

PURPOSE: To investigate the current practices in anatomy teaching at French universities in 2023. METHODS: On January 10, 2023, a questionnaire was sent to all members of the official list of the French Medical College of Professors of Anatomy. Each Anatomy centre was asked to complete this online form only once. The questionnaire covered several key themes, including broad questions, dissections practices, "virtual" dissections, teaching methods and teaching staff. RESULTS: The questionnaire was completed by 26/28 anatomy centres. Access to body donor dissection is reported to be mandatory in 15 of the 26 centers (58%), optional in 10 centers (38%), and "tolerated" in one center (4%). Fifteen of 26 centers (58%) reported having a virtual dissection table for teaching anatomy. Concerning the teaching of anatomy via live ultrasound, 10 out of 26 centers (38%) reported providing it. Regarding the teaching methods used for medical students, chalk and board lectures are the most common method, although the intensity of use varies. Most lectures are given with chalk and board in 42% (11/26) of the centers. In about 73% (19/26) of the centers, tablet lectures are used. Regarding anatomy teachers, it was reported that in 24/26 anatomy centres (92%), more than 50% of the courses for medical students are taught by professors holding the chair of anatomy (21/26 professors (81%), 3/26 associate professors (12%)). CONCLUSION: The present study endeavors to contribute to the existing body of knowledge on anatomy education by offering insights into the current practices in French universities.

The weepy cry - short neural signal bursts in intraoperative neuromonitoring.

PURPOSE: This study aimed to establish an in-vitro alternative to existing in-vivo systems to analyze nerve dysfunction using continuous neuromonitoring (C-IONM). METHODS: Three hundred sixty-three recurrent laryngeal nerves (RLN) (N(pigs) = 304, N(cattle) = 59) from food industry cadavers were exposed by microsurgical dissection following euthanasia. After rinsing with Ringer's lactate, they were tempered at 22 °C. Signal evaluation using C-IONM was performed for 10 min at 2 min intervals, and traction forces of up to 2N were applied for a median time of 60 s. Based on their post-traumatic electrophysiological response, RLNs were classified into four groups: Group A: Amplitude ≥ 100%, Group B: loss of function (LOS) 0-25%, Group C: ≥ 25-50%, and Group D: > 50%. RESULTS: A viable in-vitro neuromonitoring system was established. The median post-traumatic amplitudes were 112%, 88%, 59%, and 9% in groups A, B, C, and D, respectively. A time-dependent further dynamic LOS was observed during the 10 min after cessation of strain. Surprisingly, following initial post-traumatic hyperconductivity, complete LOS occurred in up to 20% of the nerves in group A. The critical threshold for triggering LOS was 2N in all four groups, resulting in immediate paralysis of up to 51.4% of the nerves studied. CONCLUSION: Consistent with in-vivo studies, RLN exhibit significant intrinsic electrophysiological variability in response to tensile forces. Moreover, nerve damage progresses even after the complete cessation of strain. Up to 20% of nerves with transiently increased post-traumatic amplitudes above 100% developed complete LOS, which we termed the "weepy cry." This time-delayed response must be considered during the interpretation of C-IONM signals.

[A Case of Resection of Leiomyosarcoma Suspected to Originate from the Right Ovarian Vein].

We present a 58-year-old female patient who underwent resection of a leiomyosarcoma arising from the right ovarian vein. She was referred to our hospital because of lower abdominal pain that had been present for 1 month prior to the visit. Ultrasound examination revealed a well-defined, smooth, lobulated, highly vascular mass(57 mm)adjacent to the distal portion of the duodenum. Contrast-enhanced computed tomography revealed the contrast enhancement mass (60 mm)located surround the right ovarian vein. In abdominal magnetic resonance image examination, the mass exhibited isointense signal on T1-weighted images, high signal on T2-weighted images, and restricted diffusion on diffusion- weighted images. We suspected primary leiomyosarcoma of the ovarian vein and proceeded with surgical intervention. On intraoperative findings, the mass was in contact with the duodenum and the inferior vena cava but dissection was easily achieved. We excised the mass together with the right ovarian vein. Pathological findings showed the mass was composed of proliferating spindle-shaped cells arranged in bundles. Some areas showed polygonal nuclear atypia and abnormal mitotic figures. Additional immunostaining showed positive for α-SMA, caldesmon, calponin, and negative for desmin, CD34, CKA1/AE3, S100. Based on the intraoperative findings, we diagnosed it as leiomyosarcoma arising of the right ovarian vein.

Techniques for Rapidly Sampling Six Crucial Organs in Adult Xenopus.

Xenopus has been a powerful model organism for understanding vertebrate development and disease for over a hundred years. While experimental analysis and dissection techniques of the embryo have been well documented, descriptions of adult Xenopus structures and organs, together with techniques for working with adults, have not been updated to take into consideration the requirements of such modern approaches as quantitative proteomics and single-cell transcriptomics. The cell-type and gene-centric perspectives require contrasting observations in embryonic stages to those in adult tissues. The organs of the larva undergo significant changes in their overall structure, morphology, and anatomical location all along the larval to adult transition, most notably during massive metamorphosis remodeling. Establishing robust standards for organ identification and dissection is crucial to ensure datasets resulting from studies performed at different laboratories can be consistent. The present protocol identifies six of the organs in the adult Xenopus, demonstrating methods for dissection and sampling of the heart ventricle, liver, fat body, pancreas, paired kidney, and skin of the adult Xenopus. Depending on the preservation methods, the dissected organs can be used for quantitative proteomics, single cell/nuclei transcriptomics, in situ hybridization, immunohistochemistry, histology, etc. This protocol aims to standardize tissue sampling and facilitate multi-lab investigations of the adult organ systems.

Unilateral identification of a rare superficial brachioulnoradial artery contributing to the superficial palmar arch and brachial artery continuing as the interosseous artery: a case report.

The occurrence of variations in human arterial branching of the upper limb has been commonly reported in peer-reviewed literature. However, the variability of upper limb arterial patterns may be underappreciated in medical practice, which can result in clinical and surgical errors. Here we report a case of a rare, unilateral arterial variation of the left upper limb of a 76-year-old Caucasian male cadaver, discovered during a routine educational dissection. Observed characteristics of the variation include a high brachial artery bifurcation into a superficial brachioulnoradial artery and brachial artery continuing as the interosseous artery and then a bifurcation of the superficial brachioulnoradial artery into a superficial radial and a superficial ulnar artery, which eventually contribute to the formation of the superficial palmar arch. The anatomical characteristics, prevalence, embryological origin, and clinical significance of the variation are discussed.

The application of flipped learning to a gross anatomy dissection course.

We implemented flipped learning for a gross anatomy dissection course and compared its effects on students' motivation and academic achievement with those of traditional dissection methods. We invited 142 first-year medical students at Chonnam National University Medical School to participate in this study. All participants engaged in traditional dissection methods in the first part of the study and flipped learning in the latter part. Medical students' motivation to learn anatomy by cadaveric dissection was measured using the ARCS (Attention, Relevance, Confidence, and Satisfaction) model. Thereafter, all students completed a written examination consisting of 96 multiple-choice questions. The students' mean motivational score regarding attention was significantly higher in association with flipped learning than with traditional learning. However, the students' mean motivational scores regarding relevance, confidence, and satisfaction were not significantly different between the methods. Additionally, the mean anatomy practice test score was significantly higher in association with flipped learning than with traditional learning. The students' motivational scores and anatomy practice test scores associated with flipped learning positively correlated with the extent of learning material completion. The students' responses indicated that flipped learning helped enhance the learning process, improve time management, reduce confusion during practice, and promote independent practice. The application of flipped learning to a cadaveric dissection course increased individual learning motivation, which improved learning activities both in and out of class, as well as academic achievement.

A comparison of student perspectives on body donation across healthcare professional programs: From prosection- to dissection-based curricula.

Donor-centered education offers students the opportunity to not only acquire visual and tactile experiences for applying anatomical knowledge but also a chance for students to reflect upon fundamental humanistic principles. The exploration of differences in students' viewpoints on body donation and the utilization of body donors in their education remains unexplored across various healthcare professional programs, which has an impact on student learning and experience. This study aimed to qualitatively examine the similarities and differences in student perspectives regarding body donation across three healthcare professional programs. One-page reflections from nursing (n = 37), physical and occupational therapy (n = 49), and medical students (n = 66) regarding their experiences in the anatomy laboratory at McGill University were collected and analyzed using a deductive approach based on themes and sub-themes outlined by Stephens et al. in 2019. Despite differences in their curricula, there were few discrepancies across the programs' reflections suggesting that donor-based learning had similar effects on each user group. Most students across the healthcare professional programs mentioned that their positive laboratory experiences motivated them to donate their bodies, extending the privilege they had to future generations. Nursing students did not reflect upon the notion that working with body donors provided unique learning experiences in the anatomy laboratory. Likewise, physical and occupational therapy (PT/OT) students did not address the importance of maintaining the privacy of body donors by covering certain body parts during lab. These findings show that prosection- and dissection-based exercises encourage reflective practices associated with these of medical ethics, highlighting an important advantage to including donor-based learning in anatomical education.

Investigating the status of whole-body donation across the United States of America.

Dissection of human body donors is a valuable part of anatomical education, research, and clinical training. In the United States, deceased human bodies are predominantly sourced through whole-body donation programs (BDPs) housed by academic institutions. Due to the lack of information regarding BDPs, the aim of this study was to gather information from US BDPs through a survey to better understand the donation process and standard operating procedures of these programs. In 2021, a Qualtrics survey was distributed to 125 BDPs and yielded responses from 72 program leaders. Collectively, these programs received more than 26,000 whole-body donations annually. Findings show that 70% typically receive enough donations to fit the needs of their institutions, 17% receive a surplus of donations, and 13% receive too few donations. Sixty-eight percent of programs permit next of kin body donation regularly or in times of need, and 44% allow next of kin to make changes to a donor's donation form after death. On average, over 85% of the registered donor population is composed of white individuals, and only 6 institutions have methods in place to promote diversity among their donor population. Overall, there is considerable variability in the operation of BDPs across the United States. These findings can be used to make recommendations about donor enrollment and program operations to ultimately improve the donation process. Future research needs to investigate the opinions and preferences of body donors along with their next of kin on the body donation process and associated policies.

Of mice (dogs, horses, sheep) and men: A novel comparative anatomy dissection course in a United Kingdom university.

At the University of Bristol, we established a novel dissection course to complement our anatomy degree. Students enrolled in this undergraduate course are trained as comparative anatomists, with equal time given to both human and veterinary anatomy. Historically, students opted to dissect either human or veterinary donors as part of the course. To fully reflect the comparative nature of the degree, the dissection course was redesigned so students could dissect both human and veterinary specimens as part of the same course. This facilitated a wide-ranging experience of anatomy, encouraging detailed knowledge of a multitude of species and allowing for multifaceted anatomy graduates to be ready for employment in a wide and competitive job market. Across three iterations of the amended version of the course, median marks ranged from 58.7% to 62.0%, with between 22 and 39 students enrolled. In comparison to the course prior to the introduction of the change, median marks ranged from 59.8% to 62.8%, with between 16 and 24 students enrolled. There was no significant difference between marks before or after the introduction of the concurrently comparative aspect. This paper describes the course, with learning materials and assessments considered, along with some reflection on its value. The course offers benefits to students by widening their perspective on anatomical knowledge and making them more equipped for the job market. It also broadens their understanding of form-function relationships. However, student feedback implied that having the choice between human or veterinary dissection was preferable, and this may outweigh the perceived benefits of the course.

Endoscopy-assisted excision of nasoglabellar dermoid cyst.

OBJECTIVE: To assess the endoscopic assisted excision of the nasoorbital dermaoid cyst. DESIGH: Case series. SETTING: Zagazig univesity hospitals. PARTICPANT: The study included patients with nasal dermoid who were operated using a local vertical incision with endoscopic assisted dissection and excision. MAIN OUTCOME MEASURES: Complete removal, complication, recurrence. RESULTS: In all patients, complete excision of the cyst was achieved with negligable blood loss. No recurrence was detected throughout the follow up. CONCLUSION: Endoscopic-assisted resection of the nasal dermoid cyst appears a safe and effective approach with small incision, precise dissection and satisfactory apparent scar with with low incidence of recurrence.

Direct Inside-Out Observation of Superficial White Matter Fasciculi in the Human Brain.

Background: Recent methodological advances in the study of the cerebral white matter have left short association fibers relatively underexplored due to their compact and juxtacortical nature, which represent significant challenges for both post-mortem post-cortex removal dissection and magnetic resonance-based diffusion imaging. Objective: To introduce a novel inside-out post-mortem fiber dissection technique to assess short association fiber anatomy. Methods: Six cerebral specimens were obtained from a body donation program and underwent fixation in formalin. Following two freezing and thawing cycles, a standardized protocol involving peeling fibers from deep structures towards the cortex was developed. Results: The inside-out technique effectively exposed the superficial white matter. The procedure revealed distinguishable intergyral fibers, demonstrating their dissectability and enabling the identification of their orientation. The assessment of layer thickness was possible through direct observation and ex vivo morphological magnetic resonance imaging. Conclusion: The inside-out fiber technique effectively demonstrates intergyral association fibers in the post-mortem human brain. It adds to the neuroscience armamentarium, overcoming methodological obstacles and offering an anatomical substrate essential for neural circuit modeling and the evaluation of neuroimaging congruence. Impact statement The inside-out fiber dissection technique enables a totally new perception of cerebral connectivity as the observer navigates inside the parenchyma and looks toward the cerebral surface with the subcortical white matter and the cortical mantle in place. This approach has proven very effective for exposing intergyral association fibers, which have shown to be much more distinguishable from an inner perspective. It gave rise to unprecedented images of the human superficial white matter and allowed, for the first time, direct observation of this vast mantle of fascicles on entire cerebral hemisphere aspects.

A protocol for isolation and culturing of mouse primary postmitotic photoreceptors and isolation of extracellular vesicles.

Here, we present a protocol for isolating and culturing mouse photoreceptors in a minimal, chemically defined medium free from serum. We describe steps for retina dissection, enzymatic dissociation, photoreceptor enrichment, cell culture, extracellular vesicles (EVs) enrichment, and EV ultrastructural analysis. This protocol, which has been verified for cultured cells derived from multiple murine strains, allows for the study of several aspects of photoreceptor biology, including EV isolation and nanotube formation. For complete details on the use and execution of this protocol, please refer to Kalargyrou et al. (2021).1.

Protocol to examine immune subpopulations in murine conjunctiva and lacrimal gland using flow cytometry.

Here, we present a protocol for the examination of immune cells in the murine conjunctiva and lacrimal gland using flow cytometry. We describe steps for dissection, preparation of high-quality single-cell suspensions, utilization of comprehensive staining panels, and optimization of flow cytometry voltage. We then detail procedures for compensation adjustments and the implementation of effective gating strategies. For complete details on the use and execution of this protocol, please refer to Ma et al.1.

Robotic harvest of the free gracilis muscle flap.

The conventional approach to harvest of the gracilis muscle flap necessitates a medial thigh incision that is often related to several donor site complications. In this report we describe the robotic harvest of the free gracilis muscle flap in order to reduce the morbidity associated with the open incision. Through three ports, the robotic system (Da Vinci Xi, Intuitive Surgical) enables precise dissection of the gracilis muscle, the vascular pedicle, and the obturator nerve; thus, enhancing surgical control, optimizing visualization through magnification, aiding in detailed vascular pedicle dissection and minimizing human error. We believe that the technique of totally robotic harvest of the free gracilis muscle flap, herein introduced, is a feasible and effective approach, and confers specific advantages over traditional harvest technique.

Educational stereoscopic representation of a step-by-step brain white fiber dissection according to Klingler's method.

BACKGROUND: Understanding and teaching the three-dimensional architecture of the brain remains difficult because of the intricate arrangement of grey nuclei within white matter tracts. Although cortical area functions have been well studied, educational and three-dimensional descriptions of the organization of deep nuclei and white matter tracts are still missing. OBJECTIVE: We propose herein a detailed step-by-step dissection of the lateral aspect of a left hemisphere using the Klingler method and provide high-quality stereoscopic views with the aim to help teach medical students or surgeons the three-dimensional anatomy of the brain. METHODS: Three left hemispheres were extracted and prepared. Then, according to the Klingler method, dissections were carried out from the lateral aspect. Photographs were taken at each step and were modified to provide stereoscopic three-dimensional views. RESULTS: Gray and white structures were described: cortex, claustrum, putamen, pallidum, caudate nucleus, amygdala; U-fibers, external and internal capsules, superior longitudinal fasciculus, frontal aslant fasciculus, uncinate fasciculus, inferior fronto-occipital fasciculus, inferior longitudinal fasciculus, corticospinal fasciculus, corona radiata, anterior commissure, and optic radiations. CONCLUSION: This educational stereoscopic presentation of an expert dissection of brain white fibers and basal ganglia would be of value for theoretical or hands-on teaching of brain anatomy; labeling and stereoscopy could, moreover, improve the teaching, understanding, and memorizing of brain anatomy. In addition, this could be also used for the creation of a mental map by neurosurgeons for the preoperative planning of brain tumor surgery.