The innervation of human muscularis mucosae: an ultrastructural study

The muscularis mucosae, a thin band of smooth muscle located at the base of the gastrointestinal mucosa, has been the topic of very few studies .The muscularis mucosae might regulate the absorptive and secretory functions of the gut through movements of the mucosal surface (1).The autonomic innervation of this tissue is almost completely unknown.Therefore we have carried out an ultrastructural study on nerve fibers of muscolaris mucosae by using archived mucosal rectal biopsies of children of different age , examined in the past for the diagnosis of neurometabolic disease and resulted negative. Nerve fibers of muscularis mucosae were unmyelinated. They contain several axons with the characteristics of intervaricose tract completely or almost surrounded by Schwann cells . Other axons in the nerve fibers appear as varicosities partly covered with Schwann cell cytoplasm or naked, and filled with vesicles and mitochondria .The vesicles in the same varicosity appear pleomorphic: small clear-core vesicles , dense -core of small diameter or less often dense-core of larger type. The membrane of muscle cells often protruded toward the varicosity . No synaptic specialization was observed.With very low frequency we found varicosities in intimate contact with the plasmalemma . Pleomorphic vesicles inside the same varicosity suggest a complex neurotransmission based on the release of classical transmitter and cotransmitters.The physiological relevance of these nerves remains unclear. Strips of longitudinal muscularis mucosae isolated from the human, guinea pig and rat colon responded with concentration-dependent contractions to the application of several spasmogens (1). In the human muscularis mucosae, neurokinin A was most potent, followed by carbachol, prostaglandin F2 and acetylcholine. These findings suggest the possibility that the muscularis mucosae is innervated by excitatory cholinergic nerves (1).On the other hand in oesophagus exogenously applied adrenaline inhibited spontaneous activities of the muscularis mucosae motor activity. Adrenergic nerves might inhibit spontaneous motility via the inhibition of cholinergic neurotransmission. VIP- , NPY-, CGRP- and galanin-immunoreactive nerve fibers were observed in the human esophageal muscularis mucosae but their function remains unknown (2)

The innervation of human muscularis mucosae: an ultrastructural study

The muscularis mucosae, a thin band of smooth muscle located at the base of the gastrointestinal mucosa, has been the topic of very few studies .The muscularis mucosae might regulate the absorptive and secretory functions of the gut through movements of the mucosal surface (1).The autonomic innervation of this tissue is almost completely unknown.Therefore we have carried out an ultrastructural study on nerve fibers of muscolaris mucosae by using archived mucosal rectal biopsies of children of different age , examined in the past for the diagnosis of neurometabolic disease and resulted negative. Nerve fibers of muscularis mucosae were unmyelinated. They contain several axons with the characteristics of intervaricose tract completely or almost surrounded by Schwann cells . Other axons in the nerve fibers appear as varicosities partly covered with Schwann cell cytoplasm or naked, and filled with vesicles and mitochondria .The vesicles in the same varicosity appear pleomorphic: small clear-core vesicles , dense -core of small diameter or less often dense-core of larger type. The membrane of muscle cells often protruded toward the varicosity . No synaptic specialization was observed.With very low frequency we found varicosities in intimate contact with the plasmalemma . Pleomorphic vesicles inside the same varicosity suggest a complex neurotransmission based on the release of classical transmitter and cotransmitters.The physiological relevance of these nerves remains unclear. Strips of longitudinal muscularis mucosae isolated from the human, guinea pig and rat colon responded with concentration-dependent contractions to the application of several spasmogens (1). In the human muscularis mucosae, neurokinin A was most potent, followed by carbachol, prostaglandin F2 and acetylcholine. These findings suggest the possibility that the muscularis mucosae is innervated by excitatory cholinergic nerves (1).On the other hand in oesophagus exogenously applied adrenaline inhibited spontaneous activities of the muscularis mucosae motor activity. Adrenergic nerves might inhibit spontaneous motility via the inhibition of cholinergic neurotransmission. VIP- , NPY-, CGRP- and galanin-immunoreactive nerve fibers were observed in the human esophageal muscularis mucosae but their function remains unknown (2)

Sternal foramina : anatomy and clinical significance

Vengono presentati casi di forami sternali multipli e viene discussa l'importanza della conoscenza da parte dei clinici di questa anomalia

Osteologic topography of the trochlear spine and fovea as landmarks to locate the superior oblique trochlea

The position of the superior oblique tendon, attached to the orbital roof by a cartilaginous trochlea, is marked by osteologic landmarks like the trochlear spine and/or fovea, approximately located at the superomedial angle of the orbit. Aim of the study is to place the trochlea within the orbit with a series of measurements to give the surgeon detailed references of the trochlea location. For this purpose, we undertook the study of a collection of dry skulls of known sex and age to investigate bony landmarks. Measurements were taken to assess the position of the trochlear spine/fovea on a frontal plane employing a system of vertical and horizontal lines passing through known bony reference points. Measurements were also recorded between the trochlear spine/fovea and the orbital opening on one side and the anterior rim of the optic canal on the other side. The distances of the trochlear spine/fovea from the lines passing along the supraorbital notch and the frontozygomatic suture were respectively 8.5 ± 2.3 mm and 5.7 ± 1.5 mm. The distances of the trochlear fovea/spine from the anterior orbital opening and from the anterior rim of the optic foramen were respectively 4.2 ± 0.11 and 37.5 ± 3.1 mm. Only the distance from the optic canal showed sex-related differences. In conclusion, to avoid unwanted injuries of the trochlea of the superior oblique in surgery of the superomedial angle of the orbit, the surgeon should be aware of its precise position

HISTOPATHOLOGICAL FINDINGS IN SYSTEMIC SCLEROSIS-RELATED MYOPATHY: FIBROSIS AND MICROANGIOPATHY

Objectives: The objective of this study was to identify specific histopathological features of skeletal muscle involvement in systemic sclerosis (SSc) patients. Methods: A total of 35 out of 112 SSc-patients (32%, including 81% female and 68% diffuse scleroderma) presenting clinical, biological and electromyographic (EMG) features of muscle weakness, were included. Patients underwent vastus lateralis biopsy, assessed for individual pathologic features including fibrosis [type I collagen (Coll-I), transforming growth factor β (TGF-β)], microangiopathy [cluster of differentiation 31 (CD31), pro-angiogenic vascular endothelial growth factor A (VEGF-A), anti-angiogenic VEGF-A165b], immune/ inflammatory response [CD4, CD8, CD20, human leucocyte antigens ABC (HLA-ABC)], and membranolytic attack complex (MAC). SSc biopsies were compared with biopsies of (n = 35) idiopathic inflammatory myopathies (IIMs) and to (n = 35) noninflammatory myopathies (NIMs). Ultrastructural abnormalities of SSc myopathy were also analyzed by transmission electron microscopy (TEM). Results: Fibrosis in SSc myopathy (81%) is higher compared with IIM (32%, p < 0.05) and with NIM (18%, p < 0.05). Vascular involvement is dominant in SSc muscle (92%), and in IIM (78%) compared with NIM (21%, p < 0.05). In particular, CD31 shows loss of endomysial vessels in SSc myopathy compared with IIM (p < 0.05) and with NIM (p < 0.01). VEGF-A is downregulated in SSc myopathy compared with IIM (p < 0.05) and NIM (p < 0.05). Conversely, VEGF-A165b is upregulated in SSc myopathy. The SSc immune/inflammatory response suggested humoral process with majority (85%) HLA-ABC fibral neoexpression and complement deposits on endomysial capillaries MAC, compared with IIM (p < 0.05), characterized by CD4+/CD8+/B-cell infiltrate, and NIM (p < 0.05). TEM analysis showed SSc vascular alterations consisting of thickening and lamination of basement membrane and endothelial cell ‘swelling’ coupled to endomysial/perimysial fibrosis. Conclusions: Fibrosis, microangiopathy and humoral immunity are predominant in SSc myopathy, even if it is difficult to identify specific histopathological hallmarks of muscle involvement in SSc, since they could be present also in other (IIM/NIM) myopathies. © 2016, © The Author(s), 2016

Teaching in schools of specialization: problems and the possible solutions

Teaching of anatomy in post-graduate schools that request it is particularly difficult for the number of hours available, the need not to repeat arguments already addressed in the degree course in medicine, to stimulate the interest of doctors in training and provide anatomical knowledge which are not detached from the clinical practice. To overcome these difficulties we have used in the teaching of anatomy of the post-graduate schools of the neurological-neurosurgical areas and of laryngology-phoniatry a didactic approach, which illustrate, verified for its effectiveness with an evaluation questionnaire submitted to the doctors in training at the end of the course. The essential points of the teaching are: monographic lectures on topics of anatomy related to the clinical field of specific specialization. Treatment of the subjects starting from neurological syndromes or complex brain functions of clinical relevance the understanding of which involves learning of a set of anatomical structures (eg language and cranial nerve, paralytic syndromes associated of the cranial nerves etc).The educational cycle is completed inviting the doctors to present to colleagues and to the professor the anatomical correlates of a published case report, provided to them at the end of the lesson. The teaching of the anatomy that we have illustrated is different from that which is evident from the texts available of clinical neuroanatomy, which treated anatomy of brain regions or of functional systems and reported medical cases that seek to exercise the clinical reasoning ,which purpose is not relevant to the teaching of anatomy .In conclusion even if our didactic approach is limited to some medical specializations and tested on a small number of doctors in training we suggest it as an alternative way to teach anatomy in postgraduate schools

Human osteoarthritic chondrocytes exposed to extremely low-frequency electromagnetic fields (ELF) and therapeutic application of musically modulated electromagnetic fields (TAMMEF) systems: a comparative study.

Osteoarthritis (OA) is the most common joint disease, characterized by matrix degradation and changes in chondrocyte morphology and metabolism. Literature reported that electromagnetic fields (EMFs) can produce benefits in OA patients, even if EMFs mechanism of action is debated. Human osteoarthritic chondrocytes isolated from femoral heads were cultured in vitro in bidimensional (2-D) flasks and in three-dimensional (3-D) alginate beads to mimic closely cartilage environment in vivo. Cells were exposed 30 min/day for 2 weeks to extremely low-frequency electromagnetic field (ELF) with fixed frequency (100 Hz) and to therapeutic application of musically modulated electromagnetic field (TAMMEF) with variable frequencies, intensities, and waveforms. Cell viability was measured at days 7 and 14, while healthy-cell density, heavily vacuolized (hv) cell density, and cluster density were measured by light microscopy only for 3-D cultures after treatments. Cell morphology was observed for 2-D and 3-D cultures by transmission electron microscopy (TEM). Chondrocyte exposure to TAMMEF enhances cell viability at days 7 and 14 compared to ELF. Light microscopy analysis showed that TAMMEF enhances healthy-cell density, reduces hv-cell density and clustering, compared to ELF. Furthermore, TEM analysis showed different morphology for 2-D (fibroblast-like) and 3-D (rounded shape) cultures, confirming light microscopy results. In conclusion, EMFs are effective and safe for OA chondrocytes. TAMMEF can positively interfere with OA chondrocytes representing an innovative non-pharmacological approach to treat OA

Alternative Pathways of Cancer Cell Death by Rottlerin: Apoptosis versus Autophagy

Since the ability of cancer cells to evade apoptosis often limits the efficacy of radiotherapy and chemotherapy, autophagy is emerging as an alternative target to promote cell death. Therefore, we wondered whether Rottlerin, a natural polyphenolic compound with antiproliferative effects in several cell types, can induce cell death in MCF-7 breast cancer cells. The MCF-7 cell line is a good model of chemo/radio resistance, being both apoptosis and autophagy resistant, due to deletion of caspase 3 gene, high expression of the antiapoptotic protein Bcl-2, and low expression of the autophagic Beclin-1 protein. The contribution of autophagy and apoptosis to the cytotoxic effects of Rottlerin was examined by light, fluorescence, and electron microscopic examination and by western blotting analysis of apoptotic and autophagic markers. By comparing caspases-3-deficient (MCF-73def) and caspases-3-transfected MCF-7 cells (MCF-73trans), we found that Rottlerin induced a noncanonical, Bcl-2-, Beclin 1-, Akt-, and ERK-independent autophagic death in the former- and the caspases-mediated apoptosis in the latter, in not starved conditions and in the absence of any other treatment. These findings suggest that Rottlerin could be cytotoxic for different cancer cell types, both apoptosis competent and apoptosis resistant

Muscle pathology patterns in possibly adjuvant related autoimmune/inflammatory syndrome (ASIA)

Growing evidence shows a link for biologically inert molecules, such as vaccine adjuvants and silicone implants, with the occurrence of autoimmunity-related disorders, defined as autoimmune/inflammatory syndrome induced by adjuvant-ASIA (1). Clinical conditions encompass siliconosis, the Gulf war syndrome, the macrophagic myofasciitis syndrome (MMF), post-vaccination phenomena and the spectrum of related syndromes is expanding (2). Involvement of skeletal muscle in ASIA is acknowledged in MMF, defined by long-term persistence of vaccine alum adjuvants within macrophages at sites of previous immunization. A few reports describe vaccine and silicone implants related autoimmune inflammatory myopathies (3). We carried out an immunopathological analysis of skeletal muscle biopsy in a case of MMF and two cases of possible ASIA myositis, chronologically subsequent to breast silicone implant. MMF showed the typical fascial/ perimysial macrophagic invasion, with no endomysial mononuclear infiltrates and fibral neolocalization of MHC-I complex restricted to the adjacency of macrophage deposits. The first myositis case presented with a subacute onset twenty years after an uneventful additive breast silicone implant. Endomysial inflammation, microangiopathy and multifocal fibral localization of MHC-II complex were observed. In the second patient, the onset of proximal weakness, myalgiae and a tenfold increase of creatinkinase levels occurred seven years after an unsuccessful additive mastoplasty, with rupture of prostheses and re-implantation three years later. Muscle biopsy, besides inflammation changes, showed peculiar myofibrillar disruption, with MHC-I reactive sarcoplasmic inclusions expressing several structural muscle proteins. Molecular pathogenesis of ASIA is yet undefined: genetical susceptibility is currently investigated (1,2). Due to the role of vaccines in medicine and the wide use of silicon medical devices, identification of their cause/effect link with autoimmunity is of great interest