Regulation of Vascular Smooth Muscle Cell Stiffness and Adhesion by [Ca2+]i: An Atomic Force Microscopy-Based Study.

The intracellular concentration of calcium ion ([Ca2+]i) is a critical regulator of cell signaling and contractility of vascular smooth muscle cells (VSMCs). In this study, we employed an atomic force microscopy (AFM) nanoindentation-based approach to investigate the role of [Ca2+]i in regulating the cortical elasticity of rat cremaster VSMCs and the ability of rat VSMCs to adhere to fibronectin (Fn) matrix. Elevation of [Ca2+]i by ionomycin treatment increased rat VSMC stiffness and cell adhesion to Fn-biofunctionalized AFM probes, whereas attenuation of [Ca2+]i by 1,2-Bis (2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester) (BAPTA-AM) treatment decreased the mechanical and matrix adhesive properties of VSMCs. Furthermore, we found that ionomycin/BAPTA-AM treatments altered expression of α 5 integrin subunits and α smooth muscle actin in rat VSMCs. These data suggest that [Ca2+]i regulates VSMC elasticity and adhesion to the extracellular matrix by a potential mechanism involving changing dynamics of the integrin-actin cytoskeleton axis.

An Org-1-Tup transcriptional cascade reveals different types of alary muscles connecting internal organs in Drosophila.

The T-box transcription factor Tbx1 and the LIM-homeodomain transcription factor Islet1 are key components in regulatory circuits that generate myogenic and cardiogenic lineage diversity in chordates. We show here that Org-1 and Tup, the Drosophila orthologs of Tbx1 and Islet1, are co-expressed and required for formation of the heart-associated alary muscles (AMs) in the abdomen. The same holds true for lineage-related muscles in the thorax that have not been described previously, which we name thoracic alary-related muscles (TARMs). Lineage analyses identified the progenitor cell for each AM and TARM. Three-dimensional high-resolution analyses indicate that AMs and TARMs connect the exoskeleton to the aorta/heart and to different regions of the midgut, respectively, and surround-specific tracheal branches, pointing to an architectural role in the internal anatomy of the larva. Org-1 controls tup expression in the AM/TARM lineage by direct binding to two regulatory sites within an AM/TARM-specific cis-regulatory module, tupAME. The contributions of Org-1 and Tup to the specification of Drosophila AMs and TARMs provide new insights into the transcriptional control of Drosophila larval muscle diversification and highlight new parallels with gene regulatory networks involved in the specification of cardiopharyngeal mesodermal derivatives in chordates.

Fetal Rat Gubernaculum Mesenchymal Cells Adopt Myogenic and Myofibroblast-Like Phenotypes.

PURPOSE: Gubernaculum-cremaster complex development is hormonally regulated and abnormal in a cryptorchid rat model. Using cell tracking techniques and imaging we studied myogenic phenotypes and fates in the fetal rat gubernaculum-cremaster complex. MATERIALS AND METHODS: Embryonic day 17 gubernaculum-cremaster complexes were labeled with CellTracker™ or the DNA synthesis marker EdU (5-ethynyl-2'-deoxyuridine), or immobilized in Matrigel® and grown in culture. Embryonic day 17 to 21 gubernaculum-cremaster complex sections and cells were imaged using wide field and deconvolution immunofluorescence microscopy, and muscle and/or myofibroblast specific antibodies. Deconvolved image stacks were used to create a 3-dimensional model of embryonic day 21 gubernaculum-cremaster complex muscle. RESULTS: PAX7 (paired box 7) positive and myogenin positive muscle precursors were visible in a desmin-rich myogenic zone between muscle layers that elongated and became thicker during development. Gubernaculum-cremaster complex inner mesenchymal cells expressed desmin and αSMA (α smooth muscle actin) at lower levels than in the myogenic zone. After pulse labeling with CellTracker or EdU mesenchymal cells became incorporated into differentiated muscle. Conversely, mesenchymal cells migrated beyond Matrigel immobilized gubernaculum-cremaster complexes, expressed PAX7 and fused to form striated myotubes. Mesenchymal gubernaculum-cremaster complex cell lines proliferated more than 40 passages and showed contractile behavior but did not form striated muscle. Our 3-dimensional gubernaculum-cremaster complex model had 2 orthogonal ventral layers and an arcing inner layer of muscle. CONCLUSIONS: Our data suggest that mesenchymal cells in the peripheral myogenic zone of the fetal gubernaculum-cremaster complex contribute to formation of a distinctively patterned cremaster muscle. Nonmyogenic, desmin and αSMA positive gubernaculum-cremaster complex mesenchymal cells proliferate and have a myofibroblast-like phenotype in culture. Intrinsic mechanical properties of these divergent cell types may facilitate perinatal inversion of the gubernaculum-cremaster complex.

Depletion of epidermal langerhans cells and Ia immunogenicity from tape-stripped mouse skin.

To explore the relationships among Ia antigen expression, epidermal Langerhans cells, and the immunogenicity of skin allografts, cellophane tape-stripping was used in H-2 congenic and recombinant mice of defined immunogenetic disparity. Tape-stripping of murine abdominal wall skin achieved almost complete depletion of epidermal Langerhans cells within a few hours of application, as measured by cell surface ATPase and expression of Ia antigens. Tape-stripping also reduced, to a considerable degree (but not absolutely), the Ia immunogenicity of skin allografts prepared from stripped surfaces. No comparable reduction in immunogenicity of class I major histocompatibility determinants was observed, suggesting that Langerhans cells are relatively unimportant in the presentation of H-2K antigens in skin grafts. Langerhans cells reappear within 24 h of tape-stripping to anatomically intact skin, but are detectable in orthotopically grafted only after the graft has been in residence for 4 d, i.e., shortly after it has acquired a blood supply. Repopulating Langerhans cells at that time and thereafter are exclusively of host origin. These results indicate that the traffic of Langerhans cells to the skin can be extremely dynamic, especially when the epidermal surface has been markedly disturbed, and the data imply that, under normal circumstances, large numbers of Langerhans cells can be mobilized readily from an available pool of precursors.

The structure of intersegmental muscle fibers in an insect, Periplaneta americana L.

The organization of intersegmental muscle fibers associated with the dorsal abdominal sclerites of the cockroach is described. These fibers correspond closely, in the disposition and derivation of the membranes of the transverse tubular system and sarcoplasmic reticulum cisternae, with insect synchronous flight muscle fibers, but differ markedly from these in their fibrillar architecture and mitochondrial content. The mitochondria are small and generally aligned alongside the prominent I bands of the sarcomere, and, in the best-oriented profiles of the A bands, thick filaments are associated with orbitals of twelve thin filaments, a configuration that has also been observed in striated fibers of insect visceral muscle. These structural features of insect muscles are compared and discussed in terms of possible variations in the control of contraction and relaxation, and in the nature of their mechanical role.

Isolation and characterization of myogenic precursor cells from human cremaster muscle.

Human myogenic precursor cells have been isolated and expanded from a number of skeletal muscles, but alternative donor biopsy sites must be sought after in diseases where muscle damage is widespread. Biopsy sites must be relatively accessible, and the biopsied muscle dispensable. Here, we aimed to histologically characterize the cremaster muscle with regard number of satellite cells and regenerative fibres, and to isolate and characterize human cremaster muscle-derived stem/precursor cells in adult male donors with the objective of characterizing this muscle as a novel source of myogenic precursor cells. Cremaster muscle biopsies (or adjacent non-muscle tissue for negative controls; N = 19) were taken from male patients undergoing routine surgery for urogenital pathology. Myosphere cultures were derived and tested for their in vitro and in vivo myogenic differentiation and muscle regeneration capacities. Cremaster-derived myogenic precursor cells were maintained by myosphere culture and efficiently differentiated to myotubes in adhesion culture. Upon transplantation to an immunocompromised mouse model of cardiotoxin-induced acute muscle damage, human cremaster-derived myogenic precursor cells survived to the transplants and contributed to muscle regeneration. These precursors are a good candidate for cell therapy approaches of skeletal muscle. Due to their location and developmental origin, we propose that they might be best suited for regeneration of the rhabdosphincter in patients undergoing stress urinary incontinence after radical prostatectomy.

Normal values of abdominal muscles thickness in healthy children using ultrasonography.

Abdominal muscles are one of the important elements to support the lumbar spine. Evaluation of muscle thickness using ultrasonography (US) is considered to be a source of information from muscles characteristics. The purpose of this study was to demonstrate normal reference data of abdominal muscles thickness and subcutaneous fat in adolescents using US. A random sample of 160 healthy adolescents (80 boys and 80 girls) at the age range of 15-18 years was recruited. Three abdominal muscles including Transversus Abdominis (TA), Internal Oblique (IO), External Oblique (EO) and subcutaneous fat (SF) were bilaterally measured using US. The range of normal values for TA muscle thickness was between 2.31 and 2.57 mm, for IO muscle thickness was between 4.02 and 5.15 mm and for EO muscle thickness was between 2.81 and 3.17 mm. The normal patterns of abdominal muscles were found as IO > EO > TA at both sides. Boys were taller, heavier with greater body mass index (BMI) and had larger abdominal muscles thickness than girls. A weak negative correlation was found between age and muscles size [r = (-0.06) - (-0.23), p < .05], but a significant positive correlation was found between BMI and muscle size (r = 0.21-0.68, p < .05). It seems that abdominal muscles thickness in adolescents followed the same pattern of muscle size in adults. BMI appeared to be the best predictor of muscle thickness. However, further studies are recommended to support the findings of the present study.

Neuromuscular junction in abdominal muscles of Drosophila melanogaster during adulthood and aging.

The neuromuscular junction (NMJ) of Drosophila melanogaster has been established as a productive model for the study of synaptogenesis, synaptic plasticity, vesicle recycling, and other synaptic functions in embryos and larvae. It also has potential for the study of long-term plasticity during adult life and degenerative processes associated with aging. Here we provide a detailed description of the morphology and ultrastructure of the NMJ on abdominal dorsal longitudinal muscles throughout adult life from eclosion to senescence. In contrast to the case in the larva, the predominant type of terminals in these muscles in the adult fly consists of only two or three branches with tightly packed synaptic boutons. We observed qualitative and quantitative changes as mean bouton size increased gradually during adulthood, and the largest boutons were present in the old fly. The length of nerve branches first increased and thereafter decreased gradually during most of adult life. Branch diameter also decreased progressively, but branch number did not change. The subsynaptic reticulum became progressively thinner, and "naked" boutons were found in old flies. Ultrastructural traits gave indications of an age-associated increment in autophagy, larger synaptic vesicles, and impaired endocytosis. We propose that NMJ aging in the fly correlates with impaired endocytosis and membrane dynamics. This view finds a functional correlate in flies carrying a temperature-sensitive mutation in shibire that reversible blocks endocytosis; age significantly reduces the time required for complete paralysis and increases the time of recovery, thus confirming the age-dependent alteration in vesicle dynamics.

HS1 deficiency impairs neutrophil recruitment in vivo and activation of the small GTPases Rac1 and Rap1.

Neutrophil extravasation is a critical step of the innate immune system's response to inflammation. This multistep process is tightly regulated by adhesion and signaling molecules in the endothelium and neutrophils. Activation of the ß2 integrin LFA-1 is critical for adhesion of leukocytes to postcapillary venules. This step requires coordinated activation of signaling pathways in chemokine-stimulated neutrophils, including GTPase activation and cytoskeletal remodeling, leading to conformational changes in LFA-1. Hematopoietic cell-specific lyn substrate 1 (HS1) is a cortactin-related and leukocyte-specific actin-binding protein (ABP) that regulates several processes in various immune cells. It has been shown in vitro that HS1 is important for neutrophil chemotaxis and transendothelial migration of NK cells, but its role in neutrophil extravasation in vivo has not been investigated yet. Intravital microscopy of CXCL1-stimulated cremaster venules revealed an increased rolling velocity and reduced neutrophil adhesion and transmigration in HS1 knockout (KO) mice. CXCL1-induced rapid neutrophil arrest in vivo and adhesion under flow conditions in vitro were also reduced significantly. Whereas random motility of neutrophils was unaffected, chemotaxis toward a CXCL1 gradient was reduced in the absence of HS1. Further analysis of the underlying mechanisms demonstrated that HS1 controls CXCL1-induced activation of the small GTPases Ras-related C3 botulinum toxin substrate 1 (Rac1) and Ras-related protein 1 (Rap1), thus supporting LFA-1-mediated neutrophil adhesion. Importantly, with the use of Rac1 KO neutrophils, we could show that Rac1 acts upstream of Rap1. Our results establish HS1 as an important regulator of proper Rac1 and Rap1 activation and neutrophil extravasation.

Myoblast-acellular skeletal muscle matrix constructs guarantee a long-term repair of experimental full-thickness abdominal wall defects.

To obtain a valuable treatment of congenital muscle defect, cell-matrix constructs composed of satellite cell-derived myoblasts (XY karyotype) seeded on muscle acellular matrices were used to repair a previously created full-thickness defect of abdominal wall of 18 1-month-old female Lewis rats. Acellular abdominal matrices, obtained by a detergent-enzymatic method, were positive for both basic fibroblast growth factor and transforming growth factor-beta, and were able to support in vitro cell adhesion. All animals survived the surgery, without signs of infection or implant rejection, and were humanely killed at 1, 3, or 9 months after surgery. The implants appeared well preserved, were integrated in the host tissue, and maintained their original dimension and thickness until 9 months. Vesicular acetylcholine transporter was expressed on the surface of muscle fibers from 1 month postsurgery. Finally, implanted male myoblasts were present inside the patches until 9 months, as demonstrated by the expression of SrY mRNA and by the presence of Y chromosome probe signal. These results allow us to conclude that cell-matrix constructs could represent a promising approach to the repair of muscle defects, because they are repopulated in vivo by skeletal muscle cells and nervous elements and maintain their structural integrity over the long term.

The method of isolation of the crayfish abdominal stretch receptor maintaining a connection of the sensory neuron to the ventral nerve cord ganglion.

The crayfish stretch receptor consisting of the single mechanoreceptor neurons enveloped by satellite glial cells is the simplest functioning neuroglial preparation. However, during isolation, its axons are usually transected that eliminates afferent regulation and induces complex axotomy-related signaling responses in neurons and satellite glia. We developed new microsurgical method of crayfish stretch receptor isolation, which preserves connections of sensory neurons to the ventral nerve cord ganglion. The stretch receptor may either remain on the abdominal carapace, or be completely isolated. In both cases, it may be either intact, or axotomized. The integrity of axons was confirmed by firing recording from proximal and distal axon points. Normal, necrotic and apoptotic cells were visualized using double fluorochroming with Hoechst 33342 and propidium iodide. The isolated mechanoreceptor neurons maintain regular firing during 8-10 or more hours. Glial cells surrounding non-axotomized neurons demonstrate lower necrosis and apoptosis levels than the axotomized ones. Unlike the existing method, in which the sensory neurons were axotomized, the present method preserves links between the sensory neurons and the ganglion and makes possible to avoid consequences of axotomy in neurons and satellite glia. The present neuroglial preparation may be used as a simple but informative model object in studies of axotomy-induced degeneration and survival of peripheral neurons, the role of glia in neuron injury, the signaling mechanisms of neuroglial interactions, and the effects of diverse physical and chemical factors on neuronal and glial cells.

Regional Cell-Specific Transcriptome Mapping Reveals Regulatory Complexity in the Adult Drosophila Midgut.

Deciphering contributions of specific cell types to organ function is experimentally challenging. The Drosophila midgut is a dynamic organ with five morphologically and functionally distinct regions (R1-R5), each composed of multipotent intestinal stem cells (ISCs), progenitor enteroblasts (EBs), enteroendocrine cells (EEs), enterocytes (ECs), and visceral muscle (VM). To characterize cellular specialization and regional function in this organ, we generated RNA-sequencing transcriptomes of all five cell types isolated by FACS from each of the five regions, R1-R5. In doing so, we identify transcriptional diversities among cell types and document regional differences within each cell type that define further specialization. We validate cell-specific and regional Gal4 drivers; demonstrate roles for transporter Smvt and transcription factors GATAe, Sna, and Ptx1 in global and regional ISC regulation, and study the transcriptional response of midgut cells upon infection. The resulting transcriptome database (http://flygutseq.buchonlab.com) will foster studies of regionalization, homeostasis, immunity, and cell-cell interactions.

Tissue response to polypropylene meshes used in the repair of abdominal wall defects.

The degree of integration of biomaterials used in the repair of abdominal wall defects seems to depend upon the structure of the prosthesis. Several polypropylene (PP) prostheses are currently available which differ in the number of PP filaments, the type of weave and the porosity. The aim of this study was to evaluate the integration, adhesion formation and resistance to traction of three types of PP prostheses (Marlex, Trelex and Prolene) used in the partial or total repair of abdominal wall defects. Abdominal wall defects (7 x 5 cm) were created in 54 New Zealand rabbits involving all the tissue layers (total substitutions (TS); n = 27) or all layers excluding the parietal peritoneum (partial substitutions (PS); n = 27). The defects were repaired with PP monofilament prostheses of different weave (1 mm porosity) (Marlex, n = 18; Trelex, n = 18) or bifilament (2 mm porosity) (Prolene; n = 18). They were placed in contact on one side with subcutaneous tissue and on the other with abdominal viscera or parietal peritoneum. Animals were killed at 30, 60 and 90 days and samples of prosthesis and scar tissue processed for light and scanning microscopy. The adhesion formation with viscera was evaluated. Resistance to traction was measured with a tensiometer using strips including the prosthesis and anchorage tissue. Adhesions were detected in all the TS and in four PS. Microscopic analysis revealed total integration of the TS samples by fibrous and disorganized tissue. Prostheses used for PS were integrated by white adipose tissue with the exception of the areas around the mesh nodes and anchorage zones. The foreign body reaction could be seen as a moderate accumulation of white blood cells. Tensiometric analysis showed an increase in resistance to traction with time (P < 0.001) in each type of prosthesis, but no differences were detected (P > 0.001) between them. We concluded that: (a) the formation of adhesions was almost inhibited when the parietal peritoneum was left intact; (b) in both TS and PS, polypropylene prostheses integrated completely although the composition of the scar tissue was seem to differ; and (c) resistance to traction was similar in both TS and PS.

Functional and morphological evaluation of different polypropylene-mesh modifications for abdominal wall repair.

Modern surgical hernia repair depends increasingly on synthetic meshes for the reconstruction of the abdominal wall. Despite the undisputed advantages of the polypropylene (PP) meshes currently available (Marlex, Prolene), reports of complications after implantation are increasing. Although, serious complications such as perforation and fistula formation are rare, minor and local complaints such as seromas, misfeelings and a decreased abdominal wall mobility are observed in about one-half of the patients. In regard to the exaggerated strength of the currently available mesh modifications a reduction of the material should improve the integration of the meshes into the artificial abdominal wall. In the present study, the commercially available basic mesh Prolene has been compared to two newly constructed PP-mesh modifications with reduced amounts of PP. The modifications have gradually been adopted to the physiological requirements of abdominal wall stability and mobility by reducing the amount of PP to 64% (E-BLUE) and 24% (variant A) of the Prolene mesh (developed by ETHICON, Norderstedt, Germany). All PP-mesh variants have been implanted in a rat model and studied by 3D-photogrammetry, tensiometry, light- and electron microscopy, as well as morphometry over implantation intervals of 3, 7, 14, 21 and 90 days. The data show that current constructions of PP-meshes are oversized and definitely restrict abdominal wall mobility in the present model. Sufficient stability of the artificial abdominal wall is even guaranteed by PP-mesh modifications with a reduction of PP-quantity to about 25% of the Prolene mesh. The degree of fibrosis directly correlated with abdominal wall restriction, whereas the formation of connective tissue in the interface PP-fibre/host-issue depends on the amount and activity of the inflammatory reaction. The quantity and quality of inflammation, again, directly relies to the amount of PP and to the surface area in contact with the recipient tissues. Altogether, the present study suggests that a modification of the PP-meshes could be helpful to prevent major and minor complications of surgical PP-meshes.

Influence of polyglactin-coating on functional and morphological parameters of polypropylene-mesh modifications for abdominal wall repair.

Regarding oversized mechanical properties of most of the currently available materials a new mesh was developed (ETHICON, Norderstedt, Germany) and exactly adopted to the physiology of the human abdominal wall by reducing the amount of polypropylene (weight of <30 g/m2; mesh A). The consecutive increase of pores size as well as the use of multifilaments led to a pronounced increase of flexibility. To improve the handling during operation the initial stiffness of this low-weight large pores mesh was increased by strengthening with different amounts of absorbable polyglactin (combination of glycolide and lactide) in various forms: by coating (mesh B), adding multifilament polyglactin filaments (mesh C, Vypro) or both (mesh D), respectively. To test the consequences of the different supplementary techniques all mesh variants are implanted in a rat model. Over implantation intervals of 3, 7, 14, 21 and 90 days we measured the tensile strength, the resulting stiffness and surveyed the tissue response, particularly in regard to the extent of inflammation and to the induced fibrosis. The results proved a sufficient mechanical stability of the material reduced and pure polypropylene mesh A without restriction of the mobility of the abdominal wall compared with a group that had simple laparotomy and closure. The histological analysis of the interface showed a minor inflammatory reaction and a dense vascularisation. The addition of polyglactin multifilaments (mesh C) reduces the number of macrophages and granulocytes as indicators for acute inflammation, showing generally a scar formation limited merely to the perifilamentary region. The abdominal wall compliance remained unchanged compared with mesh A. The coating of the polypropylene with polyglactin (mesh B and D) appeared to change the tissue reaction remarkably, favouring the formation of a connective tissue capsule around the whole mesh. The mechanical testing revealed an apparent protrusion with an increase of curvature of the artificial abdominal wall at rising intraabdominal pressures. The entire coating of the polypropylene surface with polyglactin induces an all embedding scar plate, filling out the pores and forming a tissue capsule. The complex interaction of tissue and implanted biomaterials with their distinct alterations of the tissue response confirms the necessity of in vivo experiments even after 'minor' modifications. Whereas the addition of polyglactin filaments appears to be favourable, the coating of polypropylene with polyglactin seems to hinder the incorporation of the mesh.

Critical evaluation of prosthetic materials in repair of abdominal wall hernias: new criteria of tolerance and resistance.

Six different materials (3 mesh and 3 cloth) commonly used for the repair of abdominal wall hernias were evaluated in 180 rats after implantation through musculature and peritoneum. Serial macroscopic and bacteriologic investigations were done, and bursting strength of the wound was determined in all groups at intervals with an original device. Histologic criteria were used to characterize the resistance of the wound and the tolerance of the host to the foreign material. Statistical analysis of the results demonstrated: (1) after postoperative day 15, the resistance of the wound was similar for each material tested; (2) during this early period mesh materials exhibited more resistance to bursting pressures than cloth materials; (3) the incorporation of mesh material was constant, whereas encystment or extrusion was always observed after implantation of cloth material; (4) no infection occurred with mesh material, but significant bacteria were found in 18 per cent of cloth material implantations; (5) the extent of the cellular reaction, the enumeration of giant, inflammatory, and fibroblastic cells showed the superiority of mesh materials; and (6) the ratio of fibroblasts to inflammatory cells reflected closely the mechanical resistance and tolerance of the foreign material.

Growth and metamorphosis of the rectus abdominis muscle in Rana pipiens.

Cross sections through the middle segment of the anuran rectus abdominis muscle were analyzed morphometrically at nine stages of development, from early larval life through full maturity. The numbers, sizes, and relative distributions of twitch and slow muscle fibers, newly differentiated fibers, degenerating fibers, and satellite cells were determined at each stage. The data indicate that the muscle increases slowly in size and fiber content during early larval life. New fibers appear to form primarily along the medial margin of the muscle. During mid-larval stages, when thyroid hormone levels are rising, new fibers form throughout the medial portion of the muscle. At a slightly later stage, fibers in the lateral region of the muscle begin to degenerate. Structurally normal presynaptic elements are present on both degenerating fibers and the empty basal laminae of fibers that had been removed by phagocytes. Both fiber formation and fiber loss slow during midmetamorphic climax, at the time when thyroid hormone levels reach a peak in anurans and begin to decline. Degenerating fibers appear within the body of the muscle at the end of metamorphosis. By the end of the second postmetamorphic month, neither degenerating nor newly differentiated fibers are present. The muscle continues to grow through adult life primarily by fiber hypertrophy.

Repair of abdominal wall defects: Gore-Tex vs. Marlex graft.

The purpose of this study was to provide experimental evidence for the role of Gore-Tex polytetraflourethylene as an abdominal wall prosthesis. This was achieved by evaluating tissue reaction in animals to the plastic and comparing it to that of Marlex mesh. Ten Wistar rats received especially prepared Gore-Tex implants, and another ten received Marlex. The materials were inserted in a fashion that yielded results both intraperitoneally and extraperitoneally. Gross and microscopic data were recorded at the time of sacrifice, which ranged from two to ten weeks postoperatively. Grossly, both plastics were found to be similar in intraperitoneal tissue reaction. Microscopically, all of the Gore-Tex grafts retained their original shape and demonstrated focal adherence to the muscle. In contrast, strands of Marlex showed disorganization in the host in 90 per cent of the specimens and no focal adherence to muscle. Instead, it was seen walled off in fibrous tissues. It was concluded that specifically formulated Gore-Tex may provide the more suitable abdominal wall prosthesis and that further research is necessary.

Surface features and behavior of the connective tissue cell.

The surface features and behavior of the connective tissue cell in normal areas and in healing incisions of the aponeuroses of the abdominal wall of the albino rat were studied with the scanning electron microscope (SEM). The normal cell is smooth, oval or triangular in outline and has a generally rounded surface and very short or no cell processes. The behavior of the fibroblasts and the condition of the fibers in and around the wounds were visualized. Shortly after the incision, the fibroblasts accumulated around the wound, possibly by increased cell division. At the site of the incision the cells responded by a self-protective mechanism in which they withdrew their processes and acquired a spheroid form. During the maximum period of a fibrillogenesis the fibroblasts enlarged, acquired a fusiform outline and long processes, exhibited a roughened surface and were surrounded with small rounded cell particles polymerizing into fine fibrils. The process of collagen secretion and three stages of collagen maturation were visualized. Some abnormal cells were seen; they could be evidence of merocrine or holocrine mode of secretion or possibly signs of cell senility.

Scanning and transmission electron microscopic study of visceral and parietal peritoneal regions in the rat.

The visceral peritoneum of intraabdominal organs (spleen, stomach, liver, small intestine), omentum majus and the parietal peritoneum of the anterior abdominal wall and the diaphragm were studied in adult Wistar rats by combined scanning and transmission electron microscopy (SEM, TEM). In general, the peritoneal surface consisted of a mesothelium composed of cubic, flat or intermediate cell types delimited by a basal lamina. Cubic mesothelial cells predominated in parenchymal organs (spleen, liver) and were characterized by prominent and indentated nuclei, a cytoplasm richly supplied with organelles, a dense microvillous coat, basal invaginations and elaborate intercellular contacts. Flat mesothelial cells were observed in the intestinal, omental and parietal peritoneum (tendinous diaphragm, abdominal wall) and showed elongated nuclei, scant cytoplasm, a poorly developed organelle apparatus and sparsely distributed microvilli. An intermediate mesothelial cell type was described within the gastric peritoneum characterized by a central cytoplasmic protrusion at the nuclear region containing most of the cytoplasmic organelles and by thin finger-like cytoplasmic processes. The submesothelial connective tissue layer was composed of collagen fiber bundles, fibroblasts and free cells (macrophages, granulocytes, mast cells) and contained blood and lymphatic vessels. In the spleen, elastic fibers formed a membranous structure with intercalated smooth muscle cells. Mesothelial openings were observed as tunnel-like invaginations within the hepatic peritoneum and as clusters of peritoneal stomata within the parietal peritoneum of the anterior abdominal wall and the muscular diaphragm. The round or oval openings of the peritoneal stomata were frequently occluded by overlapping adjacent mesothelial cells and their microvillous coat or obstructed by cellular material. At the side of the peritoneal stomata the mesothelial cell layer was interrupted to allow a direct access to the underlying submesothelial lymphatic system. The mesothelium and lymphatic endothelium shared a common basal lamina. The endothelial cells were discontinuous and displayed valve-like plasmalemmatic interdigitations facilitating an intercellular transport of fluids and corpuscular elements from the peritoneal cavity to the submesothelial lymphatic lacunae. The findings underline the morphological heterogeneity of the peritoneum in visceral and parietal regions, suggesting different functional implications, and further support the presence of extra-diaphragmatic peritoneal stomata.