Search

Purpose: In laryngeal squamous cell carcinoma (LSCC) treated with transoral laser microsurgery (TOLMS), the status of margins significantly affected local control. When a positive or close margin is present, there is no ubiquitous consensus regarding further treatments. The rationale of the present systematic review and meta-analysis is to investigate the survival impact of the status of the margins in patients affected by LSCC treated with TOLMS., Data Sources: PubMed, EMBASE, and Cochrane Library., Methods: We performed a systematic search, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Inclusion criteria were: patients affected by LSCC, staged according to the American Joint Committee on Cancer Staging System and treated by TOLMS without any previous treatment; margins status (close, positive, negative) and the adjusted hazard ratios (aHRs) with 95% confidence intervals (CIs) of overall survival, disease-specific survival, and disease-free survival has to be reported., Results: Nine studies were deemed eligible for the qualitative analysis, and 3 for the quantitative analysis to investigate the association between margin status and OS. The cumulative number of patients was 3130. The sample size ranged from 96 to 747 patients. The follow-up period ranged from 0 to 201 months. The meta-analysis results show that positive margins have an aHR of 1.30 yet with CI range (0.56 to 2.97)., Conclusions: Our current meta-analysis results are unable to definitively assess the real impact of resection margins on OS. Few authors provide accurate data regarding position and types of margins. Further prospective or high-quality studies are required., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objective: We report the management of recurrent respiratory papillomatosis (RRP) employing a protocol that includes both office-based (OB) and general anaesthesia (GA) procedures. Quality of life (QoL) outcomes in the OB cohort were compared to those obtained from an historical cohort treated only under GA., Methods: Patients affected by RRP from 2019 until 2023 ("new protocol") and from 2012 to 2019 ("historical protocol") were enrolled. In both groups the Derkay site score (DSS) was calculated. In patients adhering to the new protocol, questionnaires measuring QoL were prospectively administered (voice handicap hindex-10 [VHI-10] along with a specific questionnaire to measure the tolerance to the OB procedures). A cost analysis was also performed., Results: In all, 35 patients composed the new protocol cohort and 13 the historical. In the first group, patients underwent a median of 4 treatments. At 2 years, 68% of patients were treated exclusively in the office. Overall, for the new protocol, median DSS and VHI-10 after one year were both significantly lower than those at baseline [2 vs 4 and 3 vs 14, respectively; p < 0.001]. No differences were found between the new and the historical protocol cohorts considering DSS over time., Conclusions: Treatment of RRP may be conducted successfully in an office-based setting reducing healthcare costs., (Copyright © 2024 Società Italiana di Otorinolaringoiatria e Chirurgia Cervico-Facciale, Rome, Italy.)

Objective: The prognostic value of depth of invasion (DOI) in oral squamous cell cancer carcinoma and cutaneous melanoma is well established, while there is a lack of reports investigating the role of DOI in laryngeal cancer. This study aims to explore the association of glottic cancer DOI with other established pathological risk factors and nodal metastasis and evaluate the feasibility of measuring DOI preoperatively using tomographic imaging., Methods: The medical records of glottic cancer patients treated between 2015 and 2020 in a single tertiary referral center were screened retrospectively. Pathologically measured DOI (pDOI) value was also reviewed and registered. Preoperative computer tomography (CT) was used to obtain the radiological DOI (rDOI) measured by two dedicated radiologists. Their inter-rated agreement was assessed and the correlation between pDOI and rDOI was calculated. pDOI association with the main pathology report features was assessed with univariable analysis. Cox univariable and multivariable models were used to explore the role of pDOI on survival., Results: Ninety-one patients had pDOI data available, of which 59 also had rDOI data. A strong concordance between the two radiologists was found (concordance correlation coefficient = 0.96); rDOI and pDOI were highly and significantly correlated (R = 0.85; p < 0.001). pDOI was significantly higher in patients with perineural invasion (PNI; p < 0.001), lymphovascular invasion (LVI; p < 0.001), and nodal metastasis (p < 0.001). pDOI was associated with disease-free survival at univariable analysis (p = 0.04) while it did not show a significant impact (p = 0.10) at multivariable analysis., Conclusion: Glottic carcinoma DOI correlates with PNI, LVI, and nodal metastasis and it can be reliably assessed in a preoperative setting using CT imaging., Level of Evidence: 3 Laryngoscope, 134:3230-3237, 2024., (© 2024 The Authors. The Laryngoscope published by Wiley Periodicals LLC on behalf of The American Laryngological, Rhinological and Otological Society, Inc.)

Objective: Benign laryngeal lesions have traditionally been treated through suspension laryngoscopy under general anesthesia (GA). Recently, the development of operative videoendoscopes coupled with photoangiolytic lasers has allowed clinicians to treat these conditions in the outpatient clinic. We report our experience in the office-based (OB) setting for the treatment of patients affected by vocal fold polyps (VFPs) and Reinke's edema (RE), comparing it to patients treated under GA., Methods: A retrospective analysis was conducted on patients affected by VFP or RE. A 445 nm diode blue laser was used through the operative channel of a flexible video-endoscope for OB procedures, while GA surgeries were carried out with cold steel instrumentation. The Voice Handicap Index-10 (VHI-10) represented the primary outcome. Endoscopic outcomes, duration, and morbidity of the procedures were investigated as secondary outcomes., Results: A total of 153 patients were retrospectively enrolled. 52 were treated in an OB setting, while 91 underwent GA. Regarding patients with RE, both the OB and GA cohorts showed a significant improvement in VHI-10 (from 12.7 to 2.6 and 19.5 to 5.1, respectively; p < 0.001), as did those with VFPs (from 11.8 to 2.3 and 15.9 to 2.9 respectively; p < 0.001). No differences were found when comparing VHI-10 in the OB and GA cohorts. The mean procedural time of OB treatment (4.9 min) was significantly shorter than GA (37.1 min). No adverse events were reported., Conclusion: Our data demonstrate the efficacy and safety of the OB setting. For selected patients, OB treatments offer comparable vocal outcomes, favorable morbidity, and reduced operation times, making them an appealing alternative to the traditional approach., Level of Evidence: 3 Laryngoscope, 133:2665-2672, 2023., (© 2023 The Authors. The Laryngoscope published by Wiley Periodicals LLC on behalf of The American Laryngological, Rhinological and Otological Society, Inc.)

Studied by electron microscopy and morphometry, Auerbach's ganglia comprise nerve cell bodies that occupy ~ 40% of volume; of the neuropil, little over 30% is neural processes (axons, dendrites) and little less than 30% is glia (cell bodies, processes). The amount of surface membrane of neural elements only marginally exceeds that of glia. Glial cells extend laminar processes radially between axons, reaching the ganglion's surface with specialized membrane domains. Nerve cells and glia are tightly associated, eliminating any free space in ganglia. Glia expands maximally its cell membrane with a minimum of cytoplasm, contacting a maximal number of axons, which, with their near-circular profile, have minimal surface for a given volume. Shape of glia is moulded by the neural elements (predominantly concave the first, predominantly convex the second); the glia extends its processes to maximize contact with neural elements. Yet, a majority of axons is not reached by glia and only few are wrapped by it. Despite the large number of cells, the glia is not sufficiently developed to wrap around or just contact many of the neural elements. Mitochondria are markedly fewer in glia than in neurons, indicating a lower metabolic rate. Compactness of ganglia, their near-circular profile, absence of spaces between elements and ability to withstand extensive deformation suggest strong adhesion between the cellular elements, holding them together and keeping them at a fixed distance. Many axonal varicosities, with vesicles and membrane densities, abut on non-specialized areas of glia, suggesting the possibility of neurotransmitters being released outside synaptic sites., (© 2022. The Author(s).)

Peripheral neurons are never found alone and are invariably accompanied by glial cells, with which they are intimately associated in compact, highly deformable structures.Myenteric ganglia of the guinea-pig, examined in situ by electron microscopy, show that in their neuropil (axons and dendrites, and glial cells and processes) the glia constitutes almost half of the volume and almost half of membrane extent.In the glia, the expanse of the cell membrane predominates over that of their cytoplasm, the opposite being the case with the neural elements.The profile of the glial elements is passive and is dictated by the surrounding elements, mainly the axons, and hence it is predominantly concave.The enteric glia is widely developed; however, it is not sufficient to form a full wrapping around all neurons and around all axons (unlike what is found in other autonomic ganglia).Glial processes are radially expanding laminae, irregularly tapering, branching, and penetrating between axons.Some processes have a specialized termination attached to the basal lamina of the ganglion.Mitochondria are markedly more abundant in neural element that in the glia (up to a factor of 2).Many expanded axons, laden with vesicles clustered beneath membrane sites, abut on glial cells and processes, while these show no matching structural specializations., (© 2022. The Author(s), under exclusive license to Springer Nature Switzerland AG.)

BACKGROUND Indications for cochlear implantation (CI) are constantly being updated, and with them, the audiometric results achieved by patients. Patient satisfaction should always be considered, even in patients with lower audiological results. The aim of the present study was to compare quality of life (QoL), self-perceived hearing benefit, and audiometric results between prelingually and postlingually deafened patients, with and without sound deprivation, after CI. MATERIAL AND METHODS The sample included 46 patients with bilateral sensorineural hearing loss: 22 postlingually deafened and 24 prelingually deafened, further subdivided into sound-deprived (n=10) and non-sound-deprived (n=14). Auditory performance was evaluated with pure tone audiometry, speech recognition scores (SRS), and self-perceived hearing benefit, whereas QoL was evaluated with 2 self-reported questionnaires (Comprehensive Cochlear Implant Questionnaire and World Health Organization Quality of Life-BREF). RESULTS Audiometric results were worse in the prelingually deafened than in the postlingually deafened group, and worse in the prelingually deafened patients with sound deprivation. There was no marked difference in perceived CI benefit or QoL between the 2 groups or within the 2 prelingually deafened subgroups. No correlation was found between SRS and duration of CI use or between QoL and SRS in the prelingually and postlingually deafened groups. CONCLUSIONS Our findings demonstrate better auditory performance for the postlingually deafened group and no differences in perceived QoL or benefit of CI between the groups. The sound-deprived patients had equal scores on the perceived QoL questionnaire. These analyses suggest that sound-deprived, prelingually deafened patients may benefit from CI.

The characteristic mechanical activities of the smooth muscles found in all organs of the body are highly variable and depend mainly on the spatial arrangement of the muscle cells and the stroma: mass, orientation, relationships, links, constraints, which are deployed in various configurations. These structural features are examined here for their mechanical relevance, in light and electron microscopic views of several muscles of viscera and blood vessels, in a selection of mammalian species. Smooth muscles are incompressible and therefore maintain constant volume. They do not have available space and any movement of a part requires displacement of another part. Most of them have no terminations or points of attachment, and in hollow organs such as intestines, blood vessels and uro-genital tract they usually form structures closed onto themselves, such as rings or bag-like containers In these situations, changes in the size of the lumen is achieved very efficiently by a concentric inward enlargement that accompanies muscle contraction. The longitudinal arrangement of collagen blocks an elongation of small blood vessels upon contraction, further enhancing the efficiency of lumen reduction. In other muscles, links between layers and special arrangements of the stroma allow both shortening and elongation of a tubular organ to occur. The mechanics of smooth muscles has many characteristic features (some unique, some shared with those of hydrostats, some at variance with other muscles) and histological data are a contribution to our understanding of these properties.

To describe and illustrate the structure of the propria, the bladder of adult rats was fixed in controlled conditions of distension and examined by light and electron microscopy. The lamina propria, ~50 µm thick in the distended bladder, consists of a superficial part (the cellular component), adjacent to the urothelium, rich in nerves, capillaries, fibroblasts and thin bundles of collagen, and a deep, thicker part (the fibrous component), adjacent to the detrusor, rich in large collagen fibres and with few fibroblasts. In the cellular part there is an extensive plexus of afferent nerve fibers and a dense capillary network (with numerous pericytes), lying close to the urothelium, that is unique to the bladder. The main resident cells are fibroblasts, adhering to each other at the end of laminar extensions without forming specialized junctions. The deep part of the lamina propria is made of thick collagen fibers, interwoven and crisscrossing each other, with a few fibroblasts in the interstitial spaces between them. In summary, the superficial part of the lamina propria has most of the bladder afferent nerves, contains many fibroblasts and has a network of suburothelial capillaries. The deep part as a whole forms an ovoid balloon of woven fibrous material that is acted upon by the detrusor musculature attached to its outer surface. The lamina propria is a strong fibrous barrier between urothelium and musculature. The abundance of collagen points to the main role for its fibroblasts, that is, the production of collagen fibrils, assisting the mechanical role of the lamina propria., (© 2019 Wiley Periodicals, Inc.)

Structure and distribution of afferent nerve fibres in the rat bladder were studied by fluorescence microscopy after selective staining with antibodies against neuropeptide CGRP. Afferent fibres are very abundant (by comparison with other viscera) and interconnected in all bladder parts: muscle, urothelium, connective tissue, blood vessels, serosa. Their highest concentration is beneath the urothelium in equatorial and caudal regions, where they form a plexus, while individually maintaining a tree-like structure with innumerable branches running without preferential orientation. In cranial regions, mucosal afferent fibres become rare or absent. Abundant fibres are found in the detrusor, within each muscle bundle, with long strings of varicosities parallel to muscle cells. Afferent fibres, invariably varicose over hundreds of micrometres of their terminal parts and while still branching, comprise chains of hundreds of varicosities. Varicosities are irregular in size, frequency and separation, without specialised terminal structures around them, or within or around the fibre's ending. The possibility that varicosities are transduction points for sensory inputs is discussed, with the implication of a process taking place over considerable length in each branch of each fibre. Interconnectedness of afferent nerves of various bladder tissues, distribution of varicosities over hundreds of micrometres along axonal branches, absence of clear target structures for the fibres, apparent irregularity in the size and sequence of varicosities suggest an innervation that is not rigidly wired with distinct sensory pathways. In fact, the structural evidence suggests extensive afferent integration at the periphery, with wide distribution of source points and broad range of physical detectors.

All the cells of rat detrusor muscle fall into one of five ultrastructural types: muscle cells, fibroblasts, axons and glia, and vascular cells (endothelial cells and pericytes). The tissue is ~79% cellular and 21% non-cellular. Muscle cells occupy 72%, nerves ~4% (1/3 axons, 2/3 glia), and fibroblast >3% of space. Muscle cells (up to 6 µm across and ~600 µm long, packed to almost 100,000 per mm 2 ) have surface-to-volume ratio of 2.4 µm 2 /µm 3 ~93% of cell volume is contractile apparatus, 3.1% mitochondria and 2.5% nucleus. Cell profiles are irregular but sectional area decreases regularly towards either end of the cell. Muscle cells are gathered into bundles (the mechanical units of detrusor), variable in length and size, but of constant width. The musculature is highly compact (without fascia or capsule) with smooth outer surfaces and extensive association and adhesion between its cells. Among many types of intercellular contact and junction, digitations are very common, each muscle cell issuing minute finger-like processes that abut on adjacent cells. Sealed apposition are wide areas of specialized contact, possibly forming a chamber between two muscle cells, distinct from the extracellular space at large (stromal space). The innervation is very dense, virtually all intramuscular axons being varicose (including afferent ones). There are identifiable neuro-muscular junctions on each muscle cell, often several junctions on a single cell. There are also unattached terminals. Fibroblasts (involved in the production of collagen), ~1% of the total number of cells, do not make specialized contacts.

The spatial density of mitochondria was studied by thin-section electron microscopy in smooth muscles of bladder, iris and gut in mice, rats, guinea-pigs and sheep. Morphometric data included areas of muscle cell profiles (~6,000 muscle cells were measured) and areas of their mitochondria (more than three times as many). The visual method delivers accurate estimates of the extent of the chondrioma (the ensemble of mitochondria in a cell), measuring all and only the mitochondria in each muscle cell and no other cells. The digital records obtained can be used again for checks and new searches. Spatial density of mitochondria varies between about 2 and 10% in different muscles in different species. In contrast, there is consistency of mitochondrial density within a given muscle in a given species. For each muscle in each species there is a characteristic mitochondrial density with modest variation between experiments. On the basis of data from serial sections in the rat detrusor muscle, mitochondrial density varies very little between the muscle cells, each cell having a value close to that for the whole muscle. Mitochondrial density is different in a given muscle, e.g., ileal circular muscle, from the four mammalian species, with highest values in mouse and lowest in sheep; in mice the mitochondrial density is nearly three time higher that in sheep. In a given species there are characteristic variations between different muscles. For example, the bladder detrusor muscle has markedly fewer mitochondria than the ileum, and the iris has markedly more.

Rationale: Sinoaortic denervated (SAD) and chemically sympathectomized (SNX) rats are characterized by a decrease in arterial distensibility without hypertension and would, thus, be relevant for analyzing arterial wall stiffening independently of blood pressure level. The fibronectin network, which plays a pivotal role in cell-matrix interactions, is a major determinant of arterial stiffness. We hypothesized that in SAD and SNX rats, arterial stiffness is increased, due to alterations of cell-matrix anchoring leading to spatial reorganization of the extracellular matrix., Methods: The intrinsic elastic properties of the arterial wall were evaluated in vivo by the relationship between incremental elastic modulus determined by echotracking and circumferential wall stress. The changes of cell-extracellular matrix links in the abdominal aorta were evaluated by studying fibronectin, vascular integrin receptors, and ultrastructural features of the aorta by immunochemistry., Results: In both experimental conditions, wall stiffness increased, associated with different modifications of cell-extracellular matrix adhesion. In SAD rats, increased media cross-sectional area was coupled with an increase of muscle cell attachments to its extracellular matrix via fibronectin and its α5-β1 integrin. In SNX rats, reduced media cross-sectional area was associated with upregulation of αv-β3 integrin and more extensive connections between dense bands and elastic fibers despite the disruption of the elastic lamellae., Conclusion: In aorta of SNX and SAD rats, a similar arterial stiffness is associated to different structural alterations. An increase in αvβ3 or α5β1 integrins together with the already reported increase in the proportion of less distensible (collagen) to more distensible (elastin) components in both models contributes to remodeling and stiffening of the abdominal aorta., Competing Interests: None

Objective: In the treatment of reduced bladder capacity, matrix grafts have been used as a scaffold into which cell elements from the native bladder grow, eventually forming a new bladder segment. Functioning motor nerve endings in such segments in the rat have been demonstrated, although little is known about nerve distribution. We compare the pattern of nerve distribution in scaffold-augmented rat bladders with that in bladders regrown after subtotal cystectomy and that in control bladders., Material and Methods: Female Sprague-Dawley rats were either subtotally cystectomized (n=7) or had a part of the bladder dome replaced by an acellular collagen (small intestinal submucosa) matrix graft (n=10). Fourteen age-matched, unoperated animals were used as controls. Two and a half to 10 months after surgery the bladders were stained for acetylcholinesterase and studied in wholemounts., Results: No ganglion neurons were observed in any of the bladders. On their ventral side the control bladders showed longitudinal nerve trunks, running in parallel along the longitudinally oriented muscle bundles, while on the lateral and dorsal aspects the nerves were thinner, more irregularly arranged and frequently branched. In the bladders regrown after subtotal cystectomy, the ventral nerves were seen running obliquely to the still longitudinally oriented muscle bundles, resembling the pattern of the normal bladder base; the pattern of the dorsolateral nerves was the same as that in the controls. In the matrix bladders, the muscle and nerve patterns in the native part were the same as those in controls. Muscle bundles were growing into the matrix, accompanied by nerves, which showed limited branching when entering the matrix, usually running in parallel to the muscle, but then branching within the matrix., Conclusions: The nerves in the matrix grafts and the regrown parts of the subtotally cystectomized bladders derive from preexisting nerves in the bladder. In neither case does the nerve trunk or muscle bundle arrangement fully attain the pattern found in normal bladders.

Chronic intestinal obstruction is associated with morphological changes and functional disorders clinically reported and experimentally documented in laboratory animals. In contrast, little is known about the properties of the hypertrophied intestine after removal of the obstruction. In the present study, we removed the ileal obstruction previously applied to the ileum of rats and, after 1 or 2 weeks, studied in vitro the motor responses of de-obstructed segments of intestine to pharmacological or electrical field stimulation (EFS). By 2 weeks after de-obstruction, maximal contractile responses to receptor (acetylcholine) and non-receptor (K(+)) mediated stimuli were comparable in operated and control tissues; furthermore, the loss of sensitivity to nitric oxide (NO) unmasked in obstructed tissues was, after de-obstruction, replaced by supersensitivity to exogenous NO and vasoactive intestinal polypeptide, probably acting through cyclic nucleotide-independent pathways. Despite the complete recovery of smooth muscle responses, neurogenic contractions remained impaired in de-obstructed tissue; however, the equal contribution of cholinergic/peptidergic components to EFS responses could represent a sign of gradual but delayed recovery of enteric neurotransmission.

The superior (cranial) cervical ganglion was investigated by light microscopy in adult rats, capybaras (Hydrochaeris hydrochaeris) and horses. The ganglia were vascularly perfused, embedded in resin and cut into semi-thin sections. An unbiased stereological procedure (disector method) was used to estimate ganglion neuron size, total number of ganglion neurons, neuronal density. The volume of the ganglion was 0.5 mm3 in rats, 226 mm3 in capybaras and 412 mm3 in horses. The total number of neurons per ganglion was 18,800, 1,520,000 and 3,390,000 and the number of neurons per cubic millimetre was 36,700, 7,000 and 8,250 in rats, capybaras and horses, respectively. The average neuronal size (area of the largest sectional profile of a neuron) was 358, 982 and 800 microm2, and the percentage of volume occupied by neurons was 33, 21 and 17% in rats, capybaras and horses, respectively. When comparing the three species (average body weight: 200 g, 40 kg and 200 kg), most of the neuronal quantitative parameters change in line with the variation of body weight. However, the average neuronal size in the capybara deviates from this pattern in being larger than that of in the horse. The rat presented great interindividual variability in all the neuronal parameters. From the data in the literature and our new findings in the capybara and horse, we conclude that some correlations exist between average size of neurons and body size and between total number of neurons and body size. However, these correlations are only approximate and are based on averaged parameters for large populations of neurons: they are less likely to be valid if one considers a single quantitative parameter. Several quantitative features of the nervous tissue have to be taken into account together, rather than individually, when evolutionary trends related to size are considered., (Copyright 2004 Springer-Verlag)

The present work aims at investigating the changes in motor responsiveness of rat intestine hypertrophied by chronic mechanical obstruction. Motor responses to pharmacological agents and electrical field stimulation (EFS) were studied in hypertrophic ileal segments excised from rats subjected to experimental stenosis (n = 20) and compared with responses of control tissues from sham-operated animals (n = 20). Spontaneous motility and contractile responses to exogenous agents (KCl, acetylcholine and substance P) and EFS (10-s trains every minute, 120 mA, 0.5 ms, 1-10 Hz) were increased in hypertrophic longitudinal segments; however, normalization of motor responses to tissue wet weight revealed a remarkable reduction of contractile efficiency in hypertrophied tissues coupled with a loss of sensitivity to nitric oxide-mediated relaxation. Furthermore, EFS under non-adrenergic non-cholinergic (NANC) conditions unveiled a major role of the cholinergic component over the peptidergic one in the neurogenic contraction of hypertrophic intestine. On the whole, hypertrophic intestinal growth emerges as a dynamic process entailing adaptation of smooth muscle and neuronal structures to the increased functional load imposed by lumen obstruction.

According to the hitherto accepted view, neutrophils kill ingested microorganisms by subjecting them to high concentrations of highly toxic reactive oxygen species (ROS) and bringing about myeloperoxidase-catalysed halogenation. We show here that this simple scheme, which for many years has served as a satisfactory working hypothesis, is inadequate. We find that mice made deficient in neutrophil-granule proteases but normal in respect of superoxide production and iodinating capacity, are unable to resist staphylococcal and candidal infections. We also show that activation provokes the influx of an enormous concentration of ROS into the endocytic vacuole. The resulting accumulation of anionic charge is compensated for by a surge of K+ ions that cross the membrane in a pH-dependent manner. The consequent rise in ionic strength engenders the release of cationic granule proteins, including elastase and cathepsin G, from the anionic sulphated proteoglycan matrix. We show that it is the proteases, thus activated, that are primarily responsible for the destruction of the bacteria.

The development of the smooth musculature of viscera has attracted the interest of only relatively few investigators, and thus the field appears somewhat underexplored. The major emphasis on histochemical evidence--at the expense of ultrastructural and functional studies--may have limited the progress in this area. Mature tissue is formed through the differentiation of precursors into muscle cells and through the organization of these cells into a complex tissue where distribution and orientation of muscle cells, deployment of abundant extracellular materials and addition of other cellular elements (interstitial cells, fibroblasts, nerves, blood vessels) are characteristic and specific features. The precursor cells are found at sites where a muscle develops, and they derive predominantly from the mesoderm, but also from the neuroectoderm and from the endoderm. The process starts at different times in different organs. The earliest stages of differentiation are characterized by the precursor cells aggregating and becoming elongated; their longitudinal axis lies in a position similar to the one they will have in the mature muscle. Both the cytological and the histochemical differentiation follow distinct patterns in various muscles, with characteristic temporal sequences in the appearance of key features. This process must impart distinct functional properties to a muscle cell at each stage of its development. However, the chronological correspondence between ultrastructural and histochemical development is poorly understood. Histochemical studies have detected gradients of maturation of the muscle cells, for example, across the thickness of the gizzard musculature and along the length of the small intestine; ultrastructural studies have not yet confirmed the existence of these gradients. Muscle growth is accounted for by muscle cell enlargement (without nucleus duplication) and an increase in muscle cell number by mitosis of pre-existing differentiated muscle cells. De-differentiation and division of muscle cells, migration of muscle cells and late development of muscle cell precursors have all also been considered as possible mechanisms for muscle growth. Several authors have described the presence of precursor cells within developing smooth muscles, and they have described late differentiation of some muscle cells or waves of differentiation that would give rise to phenotypic heterogeneity of the mature muscle cell population. In contrast, other studies, mainly by electron microscopy, have suggested that, within large visceral muscles, the muscle cells differentiate synchronously. There are interesting data on the influence of adjacent tissues on the development of a smooth muscle, but the interplay of these and other factors has not been fully investigated. Smooth muscles contract from early in their development, hence mechanical factors are likely to influence development: on the one hand, passive stresses imposed on the muscle by other tissues, such as adjacent muscles or the contents of the viscera and, on the other hand, active forces generated by the muscle itself. The very attraction of visceral smooth muscles in the study of cellular morphogenesis--an attraction that has not yet been highlighted or exploited in scientific studies, either descriptively or experimentally--is that, onto a single type of cell, a large range of factors interact, such as the genetic expression, chemical influences (from other muscles, endocrine glands, nerves, other intramuscular cells) and mechanical factors.

After a surgically induced partial obstruction of the small intestine (ileum) in adult rats there is an accumulation of ingesta and a progressive enlargement of the lumen accompanied by wall thickening: over a period of 2-3 wk the circumference of the hypertrophic intestine increases by a factor of 2.7 and the thickness of the musculature increases more than threefold, while the length of the ileum (measured at the mesenteric attachment) remains unchanged. The villi become markedly larger and more elongated in the circumferential direction, and have a greater separation between one another. The number of villi per unit surface is markedly reduced but the number of villi per unit length of ileum, whilst appearing to show a small increase, was not significantly altered. The component epithelial cells (absorptive cells) appear unchanged in morphology and size (height). The microvilli of the epithelial cells have the same appearance, size (height) and packing density in the control and the hypertrophic ileum. Glands of Lieberkühn, Peyer's patches and single lymphatic follicles constituting the Peyer's patches are significantly increased in size in the hypertrophic intestine. The serosal surface of the hypertrophic ileum, in spite of the great expansion, remains regularly covered by mesothelial cells; these are much larger than in the controls and have an altered distribution of their microvilli.

The fine structure of taenia coli was studied by electron microscopy in guinea-pigs from birth to old age (over 2 years old). Smooth muscle cells are approximately 1,000 microm(3) in volume at birth, 2,200 microm(3) in young adults and 4,500 microm(3) in old age. Muscle growth and muscle cell enlargement continue throughout life, an increase in muscle volume of about 240 times. Differentiated muscle cells divide during development and in adults. Because mitoses are found in any part of the muscle, the tissue grows from within, rather than by addition at the ends or borders. There is progressive increase in nucleus volume, and decrease in surface-to-volume ratio and in nucleus-cell volume ratio in muscle cells. At all ages the taenia consists of a uniform population of muscle cells (apart from dividing cells); there are no undifferentiated cells, no precursor cells or myoblasts, and no degenerating cells. Interstitial cells and fibroblasts are observed at all ages with only small variations in relative number. The amount of intramuscular collagen increases in old age. There is roughly one capillary for every 170 muscle cell profiles at birth, and one for every 200 in adults and in old age. The innervation is dense and reaches all parts of the muscle. In adults there are approximately 1,300 axons per 10,000 microm(2) of sectional area, or between 8,000 and 38,000 axons in a full cross section of taenia; this amounts to approximately 2% of the muscle volume. An answer to the question of why there are so many nerves in the taenia was not found. Expanded axon profiles are part of typical varicose fibres. Varicosities are packed with small clear vesicles and lie at the surface of nerve bundles. Absence of strong, constant patterns indicating specialized contacts of the nerve terminals is a feature of these nerves at all ages. Some varicosities are closest to interstitial cells; more commonly they are close to muscle cells at sites that strongly suggest a neuro-muscular junction. The additional possibility that some varicosities are part of afferent fibres is discussed. The innervation is well developed at birth and the highest density of innervation is found around day 4 when 4% of the taenia consists of nervous tissue. The innervation of immature taenia is characterized by close juxtaposition of axons and muscle cells. Axon profiles packed with vesicles, varicosities and presumptive neuro-muscular junctions are present at birth. The extent of Schwann cells in intramuscular nerves is markedly less than in adults, and virtually all the axons have maximal membrane-to-membrane contact with other axons. In taenia of aged guinea-pigs, the density of innervation is reduced. There is no actual loss of nerve tissue; the total amount of nerve tissue is greater than in young adults, and the apparent reduction reflects a more intense growth of muscle cells. The Schwann cell component becomes more conspicuous than in young adults and there is a greater number of axons fully wrapped by a Schwann cell. Presumptive neuro-muscular junctions are common and probably commoner than in young adults. Growth of muscle cells, changes in their cytological features and in the stroma occur throughout life, including old age. Nerves too continue to grow and undergo structural changes in pattern of distribution, relation with Schwann cells and effector cells.

Objective: To assess the distribution and type of nerve fibers present in human peritoneal adhesions and to relate data on location and size of nerves with estimated age and with clinical parameters such as reports of chronic pelvic pain., Summary Background Data: Peritoneal adhesions are implicated in the cause of chronic abdominopelvic pain, and many patients are relieved of their symptoms after adhesiolysis. Adhesions are thought to cause pain indirectly by restricting organ motion, thus stretching and pulling smooth muscle of adjacent viscera or the abdominal wall. However, in mapping studies using microlaparoscopic techniques, 80% of patients with pelvic adhesions reported tenderness when these structures were probed, an observation suggesting that adhesions themselves are capable of generating pain stimuli., Methods: Human peritoneal adhesions were collected from 25 patients undergoing laparotomy, 20 of whom reported chronic pelvic pain. Tissue samples were prepared for histologic, immunohistochemical, and ultrastructural analysis. Nerve fibers were characterized using antibodies against several neuronal markers, including those expressed by sensory nerve fibers. In addition, the distribution of nerve fibers, their orientation, and their association with blood vessels were investigated by acetylcholinesterase histochemistry and dual immunolocalization., Results: Nerve fibers, identified histologically, ultrastructurally, and immunohistochemically, were present in all the peritoneal adhesions examined. The location of the adhesion, its size, and its estimated age did not influence the type of nerve fibers found. Further, fibers expressing the sensory neuronal markers calcitonin gene-related protein and substance P were present in all adhesions irrespective of reports of chronic abdominopelvic pain. The nerves comprised both myelinated and nonmyelinated axons and were often, but not invariably, associated with blood vessels., Conclusions: This study provides the first direct evidence for the presence of sensory nerve fibers in human peritoneal adhesions, suggesting that these structures may be capable of conducting pain after appropriate stimulation.

Adhesions in the peritoneal cavity have been implicated in the cause of intestinal obstruction and infertility, but their role in the aetiology of chronic pelvic pain is unclear. Nerves have been demonstrated in human pelvic adhesions, but the presence of pain-conducting fibres has not been established. The purpose of this study was to use an animal model to examine the growth of nerves during adhesion formation at various times following injury and to characterize the types of fibres present. Adhesions were generated in mice by injuring the surface of the caecum and adjacent abdominal wall, with apposition. At 1-8 weeks post-surgery, adhesions were processed and nerve fibres characterized histologically, immunohistochemically, and ultrastructurally. Peritoneal adhesions had consistently formed by 1 week after surgery and from 2 weeks onwards, all adhesions contained some nerve fibres which were synaptophysin, calcitonin gene-related peptide, and substance P-immunoreactive, and were seen to originate from the caecum. By 4 weeks post-surgery, nerve fibres were found to originate from both the caecum and the abdominal wall, and as demonstrated by acetylcholinesterase histochemistry, many traversed the entire adhesion. Ultrastructural analysis showed both myelinated and non-myelinated nerve fibres within the adhesion. This study provides the first direct evidence for the growth of sensory nerve fibres within abdominal visceral adhesions in a murine model and suggests that there may be nerve fibres involved in the conduction of pain stimuli., (Copyright 2000 John Wiley & Sons, Ltd.)

Increased large artery stiffness is believed to be a cardiovascular risk factor independent from mean arterial pressure. The mechanical properties of large arteries depend not only on the amounts of their main constituents (elastin, collagen, and smooth muscle cells) but also on the spatial organization and mechanical interactions among these components. These interactions may be mediated by extracellular matrix adhesion proteins and their membrane receptors or integrins. From a mechanical viewpoint, a key element may be the dense plaque, which is composed of cytoskeletal proteins linked to matrix proteins via membrane integrin receptors. Integrin expression in normal and diseased blood vessels is currently the focus of active research. In humans, hypertension-related arterial hypertrophy is not associated with an increase in intrinsic arterial wall stiffness. Aortic fibronectin expression is increased in spontaneously hypertensive rats (SHRs). By increasing cell-matrix anchoring, fibronectin may contribute to protect arterial wall components from the increased mechanical loads associated with hypertension. In atherosclerosis, the increase in cell-matrix anchoring plays a key role in preventing atheroma plaque rupture. To determine the exact role of adhesion molecules in arterial stiffness, there is a need for studies involving use of specific anti-integrin agents and of transgenic animal models.

The bladder of adult female rats receives approximately 16,000 axons (i.e., is the target of that many ganglion neurons) of which at least half are sensory. In nerves containing between 40 and 1200 axons cross-sectional area is proportional to number of axons; >99% of axons are unmyelinated. A capsule forms a seal around nerves and ends abruptly where nerves, after branching, contain approximately 10 axons. A single blood vessel is present in many of the large nerves but never in nerves of <600 axons. The number of glial cells was estimated through the number of their nuclei. There is a glial nucleus profile every 76 axonal profiles. Each glial cell is associated with many axons and collectively covers approximately 1, 000 microm of axonal length. In all nerves a few axonal profiles contain large clusters of vesicles independent of microtubules. The axons do not branch; they alter their relative position along the nerve; they vary in size along their length; none has a circular profile. All the axons are fully wrapped by glial cells and never contact each other. The volume of axons is larger than that of glial cells (55%-45%), while the surface of glial cell is twice as extensive as that of axons; there are approximately 2.27 m(2) of axolemma and approximately 4.60 m(2) of glial cell membrane per gram of nerve. Of the mitochondria of a nerve approximately 3/4 are in axons and approximately 1/4 in glial cells.