The impact of using three-dimensional digital models of human embryos in the biomedical curriculum Publication date: January 2020 Source: Annals of Anatomy - Anatomischer Anzeiger, Volume 227 Author(s): Nora Chekrouni, Roeland P. Kleipool, Bernadette S. de Bakker Abstract Background Knowledge of embryonic development is essential to understand the positioning of organs in the human body. Unfortunately, (bio)medical students have to struggle with textbooks that use static, two-dimensional (2D) schematics to grasp the intricate three-dimensional (3D) morphogenesis of the developing human body. To facilitate embryology education on an understandable and scientific level, a 3D Atlas of Human Embryology (3D Atlas) was created (Science, 2016), encompassing 14 interactive 3D-PDFs of various stages of human embryonic development (freely available from http://www.3datlasofhumanembryology.com). This study examined whether the use of the 3D atlas has added educational value and improves the students learning experience. Methods The 3D atlas was introduced and integrated in lectures and practical classes of an existing embryology course at our university for first year biomedical students. By means of a questionnaire the use of the 3D atlas was evaluated. The outcomes in written examinations was compared between cohorts that followed the course before and after integration of the 3D atlas. Results Our results showed that the 3D Atlas significantly improves students’ understanding of human embryology, reflected in significant higher test scores for new students. Furthermore, the 3D atlas also significantly improved repeaters’ test scores. Conclusions The results indicate that the3D Atlas of Human Embryology facilitates students’ learning experience as a resource to support embryology lectures. Students appreciated the use of the 3D atlas in practical classes and liked its interactive aspect. Interestingly, the students also appreciated the physical hand-painted embryological models that were used in addition to the digital 3D atlas during practical classes. The 3D Atlas of Human Embryology has proven to be a valuable resource in addition to the existing resources to teach the intricate developmental processes of human embryology, especially in a blended learning curriculum.

Onset of calciotropic receptors during the initiation of mandibular/alveolar bone formation Publication date: January 2020 Source: Annals of Anatomy - Anatomischer Anzeiger, Volume 227 Author(s): Jan Bobek, Veronika Oralova, Herve Lesot, Adela Kratochvilova, Jaroslav Doubek, Eva Matalova Abstract Mandibular/alveolar (m/a) bone, as a component of the periodontal apparatus, allows for the proper tooth anchorage and function of dentition. Bone formation around the tooth germs starts prenatally and, in the mouse model, the mesenchymal condensation turns into a complex vascularized bone (containing osteo-blasts, -cytes, -clasts) within only two days. This very short but critical period is characterized by synchronized cellular and molecular events. The m/a bone, as others, is subjected to endocrine regulations. This not only requires vasculature to allow the circulation of active molecules (ligands), but also the expression of corresponding cell receptors to define target tissues. This contribution aimed at following the dynamics of calciotropic receptors´ expression during morphological transformation of a mesenchymal condensation into the initial m/a bone structure. Receptors for all three calciotropic systemic regulators: parathormone, calcitonin and activated vitamin D (calcitriol), were localized on serial histological sections using immunochemistry and their relative expression was quantified by q-PCR. The onset of calciotropic receptors was followed along with bone cell differentiation (as checked using osteocalcin, sclerostin, RANK and TRAP) and vascularization (CD31) during mouse prenatal/embryonic (E) days 13–15 and 18. Additionally, the timing of calciotropic receptor appearance was compared with that of estrogen receptors (ESR1, ESR2). PTH receptor (PTH1r) appeared in the bone already at E13, when the first osteocalcin-positive cells were detected within the mesenchymal condensation forming the bone anlage. At this stage, blood vessels were only lining the condensation. At E14, the osteoblasts started to express the receptor for activated vitamin D (VDR). At this stage, the vasculature just penetrated the forming bone. On the same day, the first TRAP-positive (but not yet multinucleated) osteoclastic cells were identified. However, calcitonin receptor was detected only one day later. The first Sost-positive osteocytes, present at E15, were PTH1r and VDR positive. ESR1 almost copied the expression pattern of PTH1r, and ESR2 appearance was similar with VDR with a significant increase between E15 and E18. This report focuses on the in vivo situation and links morphological transformation of the mesenchymal cell condensation into a bone structure with dynamics of cell differentiation/maturation, vascularization and onset of receptors for calciotropic endocrine signalling in developing m/a bone.

Long-term treatment with naproxen changes the chemical coding of the porcine intramural duodenum neurons Publication date: January 2020 Source: Annals of Anatomy - Anatomischer Anzeiger, Volume 227 Author(s): Marta Czajkowska, Andrzej Rychlik, Jarosław Całka Abstract Due to numerous therapeutic applications and high availability, non-steroidal anti-inflammatory drugs (NSAIDs) are the most widely used drugs worldwide. However, long-term use of these drugs can lead to damage to the gastrointestinal mucosa. The enteric nervous system (ENS), which is part of the autonomic nervous system, controls most aspects of gastrointestinal activity. Enteric neurons are characterized by considerable chemical plasticity and the appearance of a pathological factor results in a change in the synthesis of neurotransmitters. The purpose of this study was to determine the effects of naproxen on expression of biologically active substances by intramural neurons supplying the porcine duodenum. The study was performed on eight immature pigs of the Pietrain x Duroc race (approximately 20 kg of body weight). The animals were divided into two groups - a control (C group) and an experimental group (N group). Group C (n = 4) consisted of animals which received empty gelatine capsules. Group N (n = 4) was composed of pigs who received naproxen orally for 28 days, approximately one hour before feeding. After this time, animals from both groups were euthanized. Frozen sections (14 μm thickness) were then prepared from the collected duodenum and subjected to double immunofluorescence staining. Antibodies against the neuronal marker PGP 9.5 and against vasoactive intestinal polypeptide (VIP), substance P (SP), neuronal nitric oxide synthase (nNOS), galanin (GAL), pituitary adenylate cyclase-activating polypeptide (PACAP) and cocaine- and amphetamine- regulated transcript peptide (CART) were used as primary antibodies. The polyclonal donkey anti-rabbit, anti-mouse and anti-guinea pig IgG antibodies - Alexa Fluor 488 and 546 - were also used for staining. Analysis of the results obtained with a fluorescence microscope showed a significant increase in the number of nNOS-, VIP-, GAL-, PACAP- and CART-immunoreactive ganglionated neurons and a decrease in the number of SP-positive neurons in the myenteric and submucosal plexuses of the porcine duodenum. The obtained results indicate the participation of enteric neurotransmitters in the neuronal duodenal response to naproxen-induced inflammation.

In vitro postovulatory oocyte aging affects H3K9 trimethylation in two-cell embryos after IVF Publication date: January 2020 Source: Annals of Anatomy - Anatomischer Anzeiger, Volume 227 Author(s): Theresa Petri, Debora Dankert, Hannah Demond, Gunther Wennemuth, Bernhard Horsthemke, Ruth Grümmer Abstract Background The physiological time axis of oocyte maturation comprises highly sensitive processes. A prolonged time span between ovulation and fertilization may impair oocyte developmental competence and subsequent embryo development, possibly due to epigenetic modifications. Since post-translational histone modifications can modify chromatin activity, and trimethylation of H3K9 (H3K9me3) has been shown to increase in the murine oocyte during maturation, here the effect of postovulatory oocyte aging on H3K9me3 was analyzed. Methods The competence of murine oocytes which were aged for 2, 4, 6 and 8 h in vitro after oocyte retrieval to develop to the two-cell and blastocyst stage was determined. Degree of H3K9me3 was analyzed in the postovulatory aged oocytes as well as in the resulting two-cell embryos after IVF. Results The current study shows that postovulatory aging of oocytes for up to eight hours after oocyte retrieval exhibited no effect on two-cell embryo and blastocyst rate; however, changes in H3K9me3 in the resulting two-cell embryos were observed. Conclusion Prolonged postovulatory oocyte aging leads to epigenetic modifications of H3K9. Such modifications may affect the developmental capacity of embryos at post-implantation developmental stages.

Anatomical study and clinical application of medial sural artery perforator flap for oral cavity reconstruction Publication date: January 2020 Source: Annals of Anatomy - Anatomischer Anzeiger, Volume 227 Author(s): Qiu-wangyue Sun, Peng-fei Gao, Chen-xing Wang, Xiao-meng Song, Xu Ding, huai-qi Li, He-ming Wu, Cheng Jie, Yu-nong Wu, Ye Yuan, jin-hai Ye Abstract The present study aims to provide anatomical evidence for clinical application of the medial sural artery perforator (MSAP) flap. The current study investigated the vascular anatomy of the flap, evaluated the postoperative appearance and function of the donor and recipient sites, and investigate the clinical value in reconstruction of oral cavity. Six lower limbs of Chinese adult cadavers were microsurgically dissected. The locations and courses of the medial sural artery perforators were identified and recorded, which provided an anatomical basis for clinical application. Then, 16 clinical cases employing this flap were evaluated, ranging from 3 × 4 cm to 6 × 8 cm, and were employed for defects in the oral cavity region. Sixteen clinical cases with intraoral soft tissue defects, which included four clinical cases with inner cheek defects, were successfully followed up for 10–47 months (24 months on average). The donor site function, contour of recipient site and oral function recovery were evaluated as acceptable or better in cases with intraoral soft tissue defect, which were further verifying the value of clinical application of MSAP in repairing oral cavity defects. Moreover, two typical clinical cases were described in detail. To conclude, the MSAP flap is a favorable choice for small- to medium-size defects based on minor donor site morbidity, satisfactory oral function recovery, perforator stability and adaptation of the pedicle for anastomosis in the oral cavity region.

Location of the myoneural junction of the inferior oblique muscle: An anatomic study Publication date: January 2020 Source: Annals of Anatomy - Anatomischer Anzeiger, Volume 227 Author(s): Marian Grace Ana-Magadia, Ma. Regina Paula Valencia, Munekazu Naito, Takashi Nakano, Hidetaka Miyazaki, Hirohiko Kakizaki, Yasuhiro Takahashi Abstract Purpose To examine both the horizontal and vertical locations of the myoneural junction of the inferior oblique muscle (IOM) in reference to the ala nasi, IOM origin, and inferior orbital rim. Materials and Methods Fifty-six orbits from 56 Japanese cadavers (15 male and 41 female cadavers; average age at time of death, 86.5 ± 9.4 years) were used in this experimental anatomical study. The inter-alae-nasi distance and the horizontal distances from the ipsilateral ala nasi to the IOM origin (ala-nasi-origin distance) and to the myoneural junction (ala-nasi-junction distance) were measured. The horizontal distance from the IOM origin to the myoneural junction (origin-junction distance) was calculated by subtraction of the ala-nasi-origin distance from the ala-nasi-junction distance. The vertical distance from the inferior orbital rim to the myoneural junction (rim-junction distance) was also measured. Results The ala-nasi-junction, origin-junction, and rim-junction distances were 12.2 ± 3.2 mm, 10.6 ± 3.2 mm, and 3.4 ± 1.0 mm, respectively. Males had a longer inter-alae-nasi distance than females (P < 0.001), although the other distances did not show statistically significant sex-related difference (P > 0.050). The ala-nasi-junction distance tended to be negatively correlated with the inter-alae-nasi distance (r = −0.222, P = 0.050). Conclusions The ala-nasi-junction distance can be affected by the inter-alae-nasi distance, which was found to be longer in males. Therefore, the IOM origin and the inferior orbital rim can be considered as more practical and reliable reference points to predict the location of the myoneural junction during the posterior inferior orbitotomy, irrespective of sex.

Virtual Special Issue „Human lymph vessels“ Publication date: Available online 13 November 2019 Source: Annals of Anatomy - Anatomischer Anzeiger Author(s): Erich Brenner

Transient connection or origin of the inferior pharyngeal constrictor during fetal development: a study using human fetal sagittal sections Publication date: Available online 11 November 2019 Source: Annals of Anatomy - Anatomischer Anzeiger Author(s): Shogo Hayashi, Hidetomo Hirouchi, Gen Murakami, Jőrg Wilting, Jose Francisco Rodríguez-Vázquez, Ji Jyun Kim Summary The inferior pharyngeal constrictor (IPC) originates from the thyroid and cricoid cartilages and inserts to the pharyngeal raphe. In serial sagittal sections of 37 embryos and fetuses at 6-15 weeks (crown rump length 15-115 mm), we found 1) the IPC connecting to the sternothyroideus and thyrohyoideus muscles (16 fetuses at 6-11 weeks) or 2) the cricothyroideus muscle (6 fetuses at 12-15 weeks) in addition to the usual cricoid origin. These aberrant connections were most likely to be transient origins of the IPC not from a hard tissue but nearby striated muscles. In four of the latter six specimens, a tendinous band from the IPC inferior end connected to the cricothyroideus muscle to provide a digastric muscle-like appearance. These aberrant connections with nearby muscles seemed to become separated by a growing protrusion of the thyroid cartilage. Therefore, these aberrant origins were, even if developed, most likely to be “corrected” to the adult morphology during midterm or late prenatal period. The aberrant or transient origin of the IPC seemed to result from a discrepancy in growth of the cartilage and muscles. Such a discrepancy in growth seems to resemble the IPC wrapping around the superior cornu of thyroid cartilage. In addition, a final or adult-like morphology was found in two of the present 37 fetal specimens. It seemed to suggest a significant redundancy in growth rate of the laryngeal structures.

Establishment of a glioblastoma in vitro (in)complete resection dual co-culture model suitable for drug testing Publication date: Available online 11 November 2019 Source: Annals of Anatomy - Anatomischer Anzeiger Author(s): Christina Schmitt, Vivian Adamski, Florian Rasch, Rainer Adelung, Ralph Lucius, Michael Synowitz, Kirsten Hattermann, Janka Held-Feindt Summary Background The treatment of glioblastomas (GBM) is still a clinical challenge. Current GBM therapeutic plans focus on the development of new strategies for local drug administration in the tumor cavity to realize an efficient long-term treatment with small side-effects. Here, different amounts of residual GBM cells and healthy brain cells define the microenvironment of the tumor cavity after individual surgical GBM resection (complete or incomplete). Methods We evaluated available in vivo data and determined the required amounts and numerical ratios of GBM and healthy brain cells for our in vitro (in)complete resection dual co-culture model. We applied a generic two-drug treatment [temozolomide (TMZ) in combination with AT101, followed by single AT101 treatment] strategy and analyzed the results in comparison with appropriate mono-culture systems to prove the applicability of our model. Results We established a suitable GBM dual co-culture model, mimicking the complete and incomplete resection in vitro, giving stable and reliable results on drug testing. Both dual co-culture conditions protectively influenced on cell death and growth rates of primary GBMs when treated with TMZ + AT101/AT101, although the treatment strategy per se was still efficient. Cell death of astrocytes correlated with amounts of increasing GBM cell numbers in the incomplete resection model upon drug treatment, and probably GBM-released chemokine and cytokines were involved in this interplay. Conclusions Our results suggest that this dual co-culture model provides a biologically relevant platform for the discovery and compound screening of local GBM treatment strategies.

Sexual dimorphism of maxillary sinuses in children and adolescents – a retrospective CT study Publication date: Available online 11 November 2019 Source: Annals of Anatomy - Anatomischer Anzeiger Author(s): Agnieszka Przystańska, Artur Rewekant, Alicja Sroka, Tomasz Gedrange, Michał Ekkert, Katarzyna Jończyk-Potoczna, Agata Czajka-Jakubowska Summary Background In the postnatal period, some sexual dimorphism is observed from the moment of birth, but it is most pronounced from puberty. The aim of the study based on CT images was to assess possible sexual dimorphism of the maxillary sinus in children aged 0-18. Methods The retrospective analysis of CT images of the head (assessed as normal by radiologists) of 170 patients aged 0 - 18 (85 females and 85 males) was performed. The maxillary sinuses of every patient were bilaterally measured in three planes. Three diameters were obtained: maximum transverse (horizontal) diameter (the maxillary sinus width, MSW); maximum vertical diameter (the maxillary sinus height, MSH) and maximum antero-posterior diameter (maxillary sinus length, MSL). The automatic CT image segmentation algorithm was used (Syngo Via for Oncology, Siemens) to establish maxillary sinus volume (MSV). The coefficient of variation used in the study allowed the biological variation between sexes to be observed. Results and conclusions The sexual dimorphism of maxillary sinus parameters is variable and depends on the phase of ontogenesis. All investigated parameters of maxillary sinuses are larger in males at 2-3 years (except for the length, which is larger in females by the end of three years, between six and nine years and after 15 years). The sexual dimorphism is less evident during the first year of age, and the most evident between 15 and 16 years. During the first two years, females are dominant with all parameters being larger than in males. The maxillary sinus length is developmentally the most stable parameter of maxillary sinuses, because throughout the investigated ontogenesis, the sexual differences are very similar. The most evident sexual dimorphism is observed in the volume of the maxillary sinus.