A new substitute for formalin: Application to embalming cadavers.

The development of formalin-free fixatives is an urgent issue in gross anatomy because of the health hazard and the tissue-hardening actions of formalin. We recently identified the fixative, antimicrobial, and preservative effects of N-vinyl-2-pyrrolidone (NVP), a precursor of the water-soluble macromolecular polymer polyvinylpyrrolidone, in animal experiments. The aim of the present study is to investigate whether NVP solution can be used as an alternative to formalin in human cadaveric dissection. Twelve donated cadavers were infused with NVP via the femoral and common carotid arteries using a peristaltic pump. Experienced teaching staff members in our department dissected the cadavers and examined their macroanatomical properties. The NVP-embalmed corpses showed no sign of decomposition or fungal growth. The bodies remained soft and flexible. Notably, the shoulder, elbow, wrist, phalangeal, hip, knee, cervical spine, and temporomandibular joints were highly mobile, almost equivalent to those of living individuals. The range of motion of most joints was greater in the NVP-fixed than formalin-fixed cadavers. Under the dermis, the subcutaneous fat was markedly reduced and the connective tissues were transparent, so the ligaments, cutaneous nerves, and veins were easily discernible. The abdominal wall and the visceral organs remained pliable and elastic, resembling those of fresh cadavers. The lungs, liver, and gastrointestinal tract were moveable in the thoracic and abdominal cavities and were readily isolated. NVP can be used successfully as a fixative and preservative solution for human cadavers; furthermore, NVP-embalmed bodies could be valuable for learning clinical skills and for training, and for developing innovative medical devices. Clin. Anat. 31:90-98, 2018. © 2017 Wiley Periodicals, Inc.

Results of Questionnaire Survey on Gross Anatomy Education (March 2014).

To understand the current situation of gross anatomy education anatomy classes. Regarding the influence of increased enrollment and to promote sharing of information on its improvement, we capacity in medical schools, many respondents were worried about conducted a questionnaire survey on gross anatomy education the impact on research activities due to the increase in teaching in September 2013. In most medical and dental schools, gross workload without expanding in teaching staff. In some schools, anatomy courses were offered to second-year students. The owing to the limitations of the facilities or the number of donated average numbers of gross anatomy practices were 34.6 in medical bodies, the number of students per cadaver had to be increased. schools and 27.4 in dental schools. The average total hours of We received various effective and practical measures for the practice in the curriculum was 125 in medical schools, and 97 improvement of gross anatomy education, such as improvement in dental schools. However, in about 80% of total schools, the of teaching materials and dissection methods, introduction of length of the actual gross anatomy practice was considerably lectures on clinical anatomy by clinicians, and implementation longer, because the students could not finish the work within of the second-round gross anatomy practice in the upper grades. the allotted class time. As to the effect of curriculum reform in Many respondents emphasized both the need for a training system respond to the introduction of the accreditation of medical and for young teaching staff, and the importance of opportunities for dental education programs, many respondents answered that sharing information on education. they had a minimal effect except earlier commencement of gross.

[Results of a questionnaire on efforts to increase research-oriented doctors].

We surveyed medical and dental schools to promote the exchange of information about university efforts to increase the number of research-oriented doctors. Periods in which students rotate through laboratories to conduct research were reported by more than two thirds of universities. Many comments asserted that these efforts are effective. However, a small number of respondents reported low student motivation and insufficient time for laboratory experience. MD-PhD courses, in which students take a leave of absence in the middle of undergraduate training and follow a PhD curriculum, have been employed by more than 10 universities. However, relatively few students have chosen such programs. Modified MD-PhD courses have recently been introduced by several universities. In these courses, by taking part of the graduate school curriculum in advance, undergraduate students can shorten the time they spend in graduate school. Students who take such courses are increasing. There were many opinions that extra positions and financial support for research-oriented doctors are effective and should be enhanced. There were also many opinions that emphasize the importance of identifying research-oriented students, improving laboratory working environments, attending academic meetings and inter-university consortia to maintain students' motivation, and promoting collaboration with departments of clinical medicine.

Formalin-free soft embalming of human cadavers using N-vinyl-2-pyrrolidone: perspectives for cadaver surgical training and medical device development.

The traditional apprenticeship approach to surgical skill education for young surgeons has drastically changed to more systematic surgical training using cadavers. Cadavers fixed with formalin are not suitable for surgical training because of their associated health hazards and overhardening. Recently, we established a formalin-free soft preservation method for human cadavers using N-vinyl-2-pyrrolidone. Since 2012, 61 cadavers have been embalmed with pyrrolidone in our institution. Tissues of pyrrolidone-embalmed cadavers are soft and pliable, and their bodies can be preserved for as long as 37 months without any signs of corruption. In this review, we introduce our recent attempts to apply pyrrolidone-embalmed cadavers in surgical and medical procedure training, including endotracheal intubation, motion physiology of the vocal folds, laparoscopic surgery, endoscopic skull base surgery, and development of novel medical devices. Future research perspectives on pyrrolidone embalming are discussed.

Piezo2 expression and its alteration by mechanical forces in mouse mesangial cells and renin-producing cells.

The kidney plays a central role in body fluid homeostasis. Cells in the glomeruli and juxtaglomerular apparatus sense mechanical forces and modulate glomerular filtration and renin release. However, details of mechanosensory systems in these cells are unclear. Piezo2 is a recently identified mechanically activated ion channel found in various tissues, especially sensory neurons. Herein, we examined Piezo2 expression and regulation in mouse kidneys. RNAscope in situ hybridization revealed that Piezo2 expression was highly localized in mesangial cells and juxtaglomerular renin-producing cells. Immunofluorescence assays detected GFP signals in mesangial cells and juxtaglomerular renin-producing cells of Piezo2GFP reporter mice. Piezo2 transcripts were observed in the Foxd1-positive stromal progenitor cells of the metanephric mesenchyme in the developing mouse kidney, which are precursors of mesangial cells and renin-producing cells. In a mouse model of dehydration, Piezo2 expression was downregulated in mesangial cells and upregulated in juxtaglomerular renin-producing cells, along with the overproduction of renin and enlargement of the area of renin-producing cells. Furthermore, the expression of the renin coding gene Ren1 was reduced by Piezo2 knockdown in cultured juxtaglomerular As4.1 cells under static and stretched conditions. These data suggest pivotal roles for Piezo2 in the regulation of glomerular filtration and body fluid balance.

Excised human larynx in N-vinyl-2-pyrrolidone-embalmed cadavers can produce voiced sound by pliable vocal fold vibration.

Tissue-hardening effect and health-hazard issue of formaldehyde (FA) have long been a great disadvantage of this conventional fixative in anatomical research. We recently developed a FA-free embalming method for cadavers which utilizes N-vinyl-2-pyrrolidone (NVP) and enables assessment of motion kinetics by maintaining the softness of embalmed tissue. By assessing the feasibility of NVP-embalmed tissue to mimic vocalization, this study aimed to prove the potential of embalmed cadavers, which have previously been used only for the understanding of anatomical morphology, for the assessment of precise motion physiology in the human body. Ten cadavers embalmed in NVP (n = 6) and FA (n = 4) were incorporated in this study. Excised larynges underwent experimental phonation to mimic vocalization with fast and pliable vibration of vocal folds. High-speed digital imaging was utilized for the assessment of vocal fold vibration. Furthermore, acoustic analysis of the voiced sound, and reproducibility examination were also performed. Regular vocal fold vibrations successfully produced voiced sounds during experimental phonation using NVP-embalmed larynges. The vibratory frequency, vibration amplitude, and stretch rate of the vocal folds were comparable to those of living humans. Six months after the first experiment, the vocal parameters were reproduced, to suggest the long-term preservation potential of our NVP-embalming technique. On the other hand, neither voiced sound nor vocal fold vibration were observed in FA-embalmed larynges. This novel embalming technique could pioneer the next era to utilize embalmed cadavers for the examination of motion physiology in the human body.

[Final report of the working group for the future planning of the Japanese Association of Anatomists].

The working group for the future planning of the Japanese Association of Anatomists (JAA) has been working to address the issues that were consulted from the president of JAA since October 2009. After making the interim report in March 2010, a public hearing for general members of the JAA was held and a final report was submitted to the President in January 2011. The report contains the analysis of the current situation, the directions in which we should proceed, and recommendations of concrete actions that JAA should take for each issue. The issues discussed were as follows: 1. Future prospects of anatomy and morphological sciences. How can we maintain the specialties of morphological and anatomical sciences in the rapidly advancing field of life sciences and develop collaborations with other fields? 2. Improvement of the JAA academic meetings. How can we increase JAA members and young participants in the academic meetings of the JAA? 3. Fostering the next generation of young researchers. How can we increase young researchers graduated from the schools of Medicine or Dentistry? 4. Future prospects of education of gross anatomy. Prospects of education in gross anatomy and the body donation registration system in relation with some new cadaver-related movements.

[Draft of guidelines for human body dissection for clinical anatomy education and research and commentary].

This article analyses the Draft of Guidelines for Human Body Dissection for Clinical Anatomy Education and Research drawn by the Study Group for Future Training Systems of Surgical Skills and Procedures established by the Fiscal Year 2010 research program of the Ministry of Health, Labor and Welfare. The purpose of the Draft of Guidelines is: First, to lay out the required basic guidelines for human cadaver usage to allow medical and dental faculty to conduct clinical education and research in accordance with existing regulations. Second, the guidelines are expected to give physicians a regulatory framework to carry out cadaver training in accordance with the current legal framework. This article explains the Draft of Guidelines in detail, outlines the future of cadaver training, and describes issues which must still be solved.

[Draft of Guidelines for Human Body Dissection for Clinical Anatomy Education and Research and commentary].

This article analyses the Draft of Guidelines for Human Body Dissection for Clinical Anatomy Education and Research drawn by the Study Group for Future Training Systems of Surgical Skills and Procedures established by the Fiscal Year 2010 research program of the Ministry of Health, Labor and Welfare. The purpose of the Draft of Guidelines is: First, to lay out the required basic guidelines for human cadaver usage to allow medical and dental faculty to conduct clinical education and research in accordance with existing regulations. Second, the guidelines are expected to give physicians a regulatory framework to carry out cadaver training in accordance with the current legal framework. This article explains the Draft of Guidelines in detail, outlines the future of cadaver training, and describes issues which must still be solved.

Morphological study of the fetal parotid duct and buccinator muscle and the relationship to salivary secretion.

The parotid glands secrete about 25% of all saliva produced. In the presence of a stimulus, the amount of saliva secreted from the parotid gland increases to 50%. A decrease in the amount of produced saliva due to aging and parotiditis results in a dry mouth. Therefore, the parotid duct is important to maintaining a healthy oral cavity. In human adults, the parotid duct, approximately 6-8-cm long, travels over the masseter muscle and penetrates the buccinator muscle to enter the oral cavity. Although there have been various studies regarding the parotid gland, only few suggest a functional role of the parotid duct, especially its area of penetration of the buccinator muscle. In this study, 34 fetal specimens ranging from 4 to 10 months of age at death were dissected for anatomical and histological examinations. The area of the parotid duct penetrating the buccinator muscle was fully formed in 5-month-old fetuses. This study found buccinator muscle fibers invading the parotid duct wall near its opening in 6-month-old fetuses and older. Our results support the claim that the buccinator muscle may act as a sphincter, playing a role in regulating and possibly preventing the reflux of salivary secretions into the parotid duct.

A new human cadaver model for laparoscopic training using N-vinyl-2-pyrrolidone: a feasibility study.

The demand for cadavers for clinical skills training is increasing. However, conventional formalin-fixed bodies are often unsuitable for surgical training because the tissues become too hard. We recently developed a new formalin-free embalming method with N-vinyl-2-pyrrolidone (pyrrolidone) that has excellent fixative, disinfectant, and preservative properties, while still keeping tissues soft and pliable. In the study reported here we investigated the feasibility of laparoscopic manipulation using pyrrolidone-fixed cadavers. Donated cadavers were embalmed either with pyrrolidone (n = 7) or with formalin-containing fixative (n = 3). A laparoscopic 12-mm trocar was inserted into the umbilical region, and CO2 gas was insufflated. Intra-abdominal structures were observed with an endoscopic camera. In the pyrrolidone-embalmed cadavers, the abdomen remained soft and depressed. In addition, CO2 injection resulted in a marked expansion of the abdominal cavity, and it was possible to move the laparoscope freely in all directions. Clear endoscopic images of the abdominal viscera were obtained. The gallbladder and rectum were identified by grasping the surrounding organs with forceps. By contrast, in the formalin-fixed bodies, the abdominal wall was rigid, and it was difficult to move the laparoscope in the peritoneal space and observe structures in detail. The amount of CO2 and changes in abdominal diameter and circumference in response to CO2 injection were significantly larger in the pyrrolidone group. In conclusion, we successfully created a sufficient pneumoperitoneum state and obtained clear endoscopic images in the pyrrolidone-embalmed cadavers. Handling and dissection of the intra-abdominal structures with forceps closely replicated real-life surgery. These findings suggest the feasibility of laparoscopic training on cadavers embalmed with this pyrrolidone fixative.

Study of morphological structure in human fetus Wharton's duct opening area.

OBJECTIVES: Not many morphological studies have been reported regarding the human salivary main duct, especially those based on observations of the fetal salivary main duct at various developmental stages. The relationships between fetal structural development and its function, functional disorders and diseases certainly have clinical significance. METHODS: In this study, we conducted a morphological observation of seven 5-9 month old human fetal sublingual caruncula, the common opening between the submandibular (Wharton's) duct and the sublingual (Bartholin's) duct. After seven specimens were removed and paraffin sections were prepared, HE and EVG staining were used for histological study of structural differences in the various developmental stages. Furthermore, anti-α-SMA staining was used to observe the distribution of the smooth muscle inside of the Wharton's duct wall. RESULTS: An abundance of elastic fibers were observed in the duct walls of both five and nine month old specimens with EVG staining. The smooth muscle fibers of five month old fetus were immature and were starting to distribute throughout the duct wall, whereas nine month old fetus' smooth muscle fibers were further developed and composed in layers. CONCLUSION: In this study, we confirmed that the Wharton's duct wall structure of nine month old fetuses was close to its adult structural formation, while the inner structure consisted of an ample distribution of elastic fibers and smooth muscle fibers. Based on the distribution pattern of the smooth muscle, we speculate that the Wharton's duct's opening area does not possess a sphincter-like function in the regulation of salivary flow.

Usefulness of N-vinyl-2-pyrrolidone Embalming for Endoscopic Transnasal Skull Base Approach in Cadaver Dissection.

Formalin or formaldehyde is commonly used for cadaver fixation, which is, however, not suitable for endoscopic transnasal skull base approach because of consequent hardening of the soft tissue. Several alternatives have been reported, but each of them also has some limitations. We applied a novel fixation method using N-vinyl-2-pyrrolidone (NVP), a precursor of the water-soluble macromolecular polymer, for endoscopic transnasal skull base approach in six donated cadaver specimens. In four cadavers, elasticity of the soft tissue in the nasal cavity was almost similar to that of living tissue, and a surgical approach similar to the real surgical field was possible. However, the soft tissue was moderately stiffer than living tissue in two specimens so that surgical manipulation was hindered to some extent while NVP concentration was 10% in all the cadavers. Since the brain tissue was too soft and pliable for surgical manipulation in NVP, more careful surgical manipulation than real surgical field was mandatory in order to prevent damage in the brain tissue. Therefore, this concentration of NVP was considered to be appropriate. In conclusion, NVP embalming was effective for endoscopic transnasal skull base approach in cadaver dissection, providing environment similar to the real surgical field.

Topographic anatomy of the subthalamic nucleus localized by high-resolution human brain atlas superimposing digital images of cross-sectioned surfaces and histological images of microscopic sections from frozen cadaveric brains.

Despite the recent advent of neuro-radiographic techniques, creating a 'perfect' human brain atlas providing precise and consistent images with minimal distortion is practically difficult. In this study, we created a new human brain atlas from cadaveric brains with serial sections of 50 µm thickness covering the entire basal ganglia. Human cerebral hemispheres were obtained from 10 donated cadavers and fixed in 10% formalin solution, cut in a block measuring 50 mm × 30 mm × 50 mm around the midpoint of the anterior and posterior commissures and frozen at -40 °C. Each block was cut into 50-µm-thick sections on the freezing microtome and the cross-sectioned surface was photographed. Simultaneously, every 10th slice from one sagittal hemisphere was sampled and stained using the Kluver-Barrera method. Prepared slides were photographed under light microscopy, and data from digital images of the cross-sectioned surface (DICSS) and digital images from microscopic sections (DIMS) were processed. Gray areas on DICSS largely represented areas of dense cellularity, and around subthalamic nucleus (STN), the zona incerta and field of Forel were clearly distinguishable on the anterosuperior side, as was the substantia nigra on the caudal side. DICSS successfully delineated the anatomical structure identical to the STN and surrounding contiguous nuclei. This new brain atlas will allow elucidation of anatomy that cannot be clearly disclosed from modern radiographic imaging or is very difficult to analyze with spatially inconsistent histological sections, and will contribute to further progress in anatomical studies of the human basal ganglia.

Identification of two accessory muscle bundles with anomalous insertions in the flexor side of the right forearm.

During dissection practice held at Kyorin University School of Medicine in 2004, two anomalous muscles were observed on the Rt-forearm-flexor-side of an 83-year-old man. The results of this investigation are reported. One accessory muscle originated from the tendinous insertion of the biceps brachii and medial epicondyle. After passing through the deep layer of the pronator teres, it became tendinous, passing towards the trapezium and second metacarpal base. Its two origins fused superficial to the ulnar artery distal to the cubital fossa, and it merged with the deep region of the pronator teres. More distally, the accessory muscle formed a belly before again becoming tendinous and bifurcated, one branch attaching to the trapezium and the other fusing with the belly of the second accessory muscle. These findings suggested that this accessory muscle was similar to Gantzer's muscle. The other accessory muscle arose distal to the origin of the flexor pollicis longus and inserted onto the second metacarpal base. In addition, from the distal side of its origin, a small muscle bundle was formed and became tendinous. It fused with the insertion tendon of the first accessory muscle to the trapezium. The second accessory muscle was thought to be deep radial carpal flexor.

Morphological classification of the anterior and middle talocalcaneal joints in terms of biomechanical stress dispersion.

Of the various foot joints, the level of interest shown in the talocalcaneal joint has been notably low. However, when considering stress dispersion during bipedal locomotion, the talocalcaneal joint plays an important role. Present study examined and morphologically classified the talocalcaneal joint. The present study investigated the morphology of the anterior talocalcaneal articular surface (A) and middle talocalcaneal articular surface (M) of the calcaneus. Using 94 feet of 47 human cadavers for anatomical dissection, the morphology of the A and M was classified into the following 6 types: Type 1, double ellipsoid, and A is markedly smaller than M; Type 2, gourd-form, and A is markedly smaller than M; Type 3, double ellipsoid, but A and M are partially in contact, Type 4, double ellipsoid, A and M are completely separated; Type 5, cylinder-form; and Type 6, others. Types 4 and 5 accounted for the majority of cases. In upright standing and bipedal locomotion, the anterior and middle talocalcaneal joints receive the anterior component of body weight dispersion in the posterior foot region. Morphological classification of articular surfaces that actually bear body weight is important, reflecting the degree of stress dispersion.

What is the true location of Jacoby's line?

We investigated 60 cadavers to examine the accurate location of conus medullaris, lower end of dura mater, and intercrestal line (Jacoby's line). Jacoby's line was established using the method described by Jacoby GW (1899). Then the intersection of this line and vertical line through the spinous processes was marked by a fine nail. After a laminectomy was performed to expose the spinal cord, spinal level of the conus medullaris, the lower end of dura mater, and Jacoby's line were examined, and marked and recorded these locations of nailing. In 2 of 48 cases (2.1%), Jacoby's line passed through the level superior to 3rd lumber vertebra (L3). The Conus medullaris was at the level beneath L3 in 2 of 57 cases (3.5%). And lower end of dura mater was located beneath the superior margin of S3 in 1 of 28 cases (3.6%). When Jacoby's line is used as a guide-point, the site for subarachnoid puncture is established in the L3-4 interspinous space. But when we consider the risk of spinal cord injury cannot be completely excluded, in case that conus medullaris is located inferior to L3 in 3.5% of cases. Conseqently, it should be important to avoid performing subarachnoid puncture at L2-3 or higher lumbar vertebral level especially in the elderly.

Establishing intercrestal line by posture:--a radiographic evaluation.

In this study, we examined the position of intercrestal line that is called Jacoby's line at heights of the vertebral body and spinous process in different positions of the lumbar spine using radiographs of 100 subjects. At the height of the spinous process, Jacoby's line was most often observed in the extension position in L4 spinous process middle 1/3 region in males and in L4-5 interspinous region in females. In the neutral position, it was in L4 spinous process middle 1/3 region in males and in L4-5 interspinous region in females. And that in the flexion position, it was in L4-5 interspinous region in both males and females. At the height of the vertebral body, Jacoby's line was most often observed in the extension position in L4-5 intervertebral region in males and in L5 vertebral body upper 1/3 region in females. In the neutral position, it was in L4 vertebral body lower 1/3 region in males and in L5 vertebral body upper 1/3 region in females. In the flexion position, it was in L5 vertebral body upper 1/3 region in males and in L5 vertebral body upper 1/3 region or L4 vertebral body lower 1/3 region in females. There was no gender difference in the most often observed position of Jacoby's line at the height of the spinous process in the flexion position. The position of Jacoby's line was found to vary in different positions of the lumbar spine. Positioning Jacoby's line in the neutral lumbar spine from the body surface, as conventionally proposed, was not always accurate, and it was considered practical to establish Jacoby's line using the spinous process in the flexion of the lumbar spine as the standard. When Jacoby's line is established by this method, it is considered to be positioned between L4-5 spinous processes middle 1/3 regions. In subarachnoid puncture in elderly patients, the puncture may be performed at a higher position in relation to the spinal cord because of spinal degeneration. Therefore, it is important to perform subarachnoid puncture in elderly patients at a site lower than the established Jacoby's line.

Morphological Analysis for Neuron-Like Cells in the Vomeronasal Organ of Human Fetuses at the Middle of Gestation.

The vomeronasal organ (VNO) of 5-month-old fetuses was examined immunohistochemically by the use of an antiserum to protein gene product 9.5 (PGP). The purpose was to identify if the human fetal VNO is lined by neuroepithelium. The PGP antiserum labeled abundant cells within the vomeronasal epithelium (VE), nerve fiber bundles in its lamina propria, and cells associated with these bundles. PGP-immunoreactive (ir) vomeronasal epithelial cells were classified into three subtypes. Type I cells, about 44% of the total cells observed, did not have any processes and tended to be located in the basal layer of the VE. Type II cells, about 37% had a single apical process that projected toward the lumen, ending at the epithelial surface. Type III cells sent a prominent process mainly toward the basement membrane, and occupied about 19% of the total cells observed. In the lamina propria, a considerable number of PGP-ir cells was observed. Some of them were present in nerve fiber bundles and contained processes parallel to the bundles. In addition, PGP-ir nerve fiber bundles and cells associated with them were even present in the portion of the nasal septal mucosa that was very close to the brain. The present results strongly suggested that the VE in human fetuses at mid-gestation is a neuroepithelium and that the VE may produce migrating cells toward the brain.

Seven skeletal muscles rich in slow muscle fibers may function to sustain neutral position in the rodent hindlimb.

Skeletal muscles consist of slow-twitch and fast-twitch muscle fibers, which have distinct physiological and biochemical properties. The muscle fiber composition determines the contractile velocity and fatigability of a particular skeletal muscle. We analyzed the systemic distribution of slow muscle fibers in all rodent skeletal muscles by myosin ATPase staining and found that only seven hindlimb skeletal muscles were extremely rich in slow muscle fibers. These included the mouse piriformis (56.5%), gluteus minimus (35.7%), vastus intermedius (24.7%), quadratus femoris (69.9%), adductor brevis (44.3%), gracilis (24.6%), and soleus muscles (35.1%). In mice, the relative proportion of slow muscle fibers did not exceed 15% in skeletal muscles in other regions. The distribution of slow muscle fibers was well conserved in rats and rabbits. The soleus muscle is an important antigravity muscle in both rodents and humans; therefore, these skeletal muscles rich in slow muscle fibers might play an important role in sustaining neutral alignment of the lower extremity.