Transport of cortisol,progesterone and cholesterol across isolated mesentery

Summary Isolated rat mesentery, mounted in a diffusion cell, is used as a model for the study of vascular endothelium permeability characteristics. The passage of tracer molecules is measured in the absence of osmotic or hydrostatic pressure gradients across the mesentery. The permeability coefficient of the membrane for cortisol and progesterone is similar. When bound to transcortin, cortisol crosses mesentery at a significantly slower rate. Metyrapone diatartrate increases by 30% the passage of free and of transcortinbound cortisol, but is without effect on the passage of progesterone or glucose in the same conditions. When the transfer of cholesterol across mesentery is studied, a high percentage of the tracer is trapped by the membrane.     

The fate and role of openings formed in the mesentery of bullfrog tadpoles,with reference to the contour of mesothelial cells

Using morphological techniques, histological changes of the mesentery were observed during the development of the bullfrog, Rana catesbeiana. The tadpoles of this species had many openings all over the mesentery from the duodenum through the large intestine. Most of the openings were elliptical and less than 3 × 2 mm in size. The openings became remarkably decreased in size and number with rapid narrowing of the mesentery occurring during the period of metamorphic climax, and had almost completely disappeared by the end of metamorphosis. Appearance and disappearance of the openings were closely correlated with the changes in the dimensions of the mesentery. Furthermore, in parallel with these changes in the openings, a noticeable alteration occurred in the shape of the mesothelial cells of the mesentery. In tadpoles having no mesenteric openings, the mesothelial cells had a polygonal contour, which became transformed once the openings were formed in the mesentery. The shapes of the transformed cells were classified into two types, one having many radiating cell processes and the other a very slender and spindle-shaped contour. Both types of cells eventually became transformed into a definitive type of cell exhibiting a roundish polygonal contour by the end of metamorphosis. From these findings it was concluded that the growing mesentery might, of necessity, give rise to the openings and transformation of the mesothelial cells to enable rapid lengthening and shortining of the intestinal tract to occur during the postembryonic development of anuran amphibians.     

Adrenergic terminal structures in the mesentery of mammals

This work is devoted to the study of adrenergic terminal structures in the mesentery of mammals (cat, dog). The investigation was performed with the Falck-Hillarp method of catecholamine fluorescence microscopy on total stretch mesentery preparations. The investigation showed that richly developed perivascular plexus constitute the basis of the adrenergic innervation system of the mesentery. In numerous points of these plexuses, single adrenergic fibers or polyaxonal structures are observed to issue into nonvascular areas of the mesentery where after repeated dichotomic division they pass into the preterminal and terminal parts. Being constructed on the principle of extended or restrained arborizations, these innervating structures have a morphological similarity with free sensory nerve endings. In this connection, the question of the possible existence of the sensory (afferent) links in the catecholamine-containing vegetative nerve plexuses is discussed.     

家兔盲肠和阑尾淋巴管的研究

用色素穿刺注入法、动脉内墨汁硝酸银水溶液注入法研究了家兔盲肠和阑尾淋巴管的起始、走向及所属淋巴结。盲肠和阑尾的毛细淋巴营均起始于固有膜,后依次穿过肠壁各层移行为系膜内淋巴管。后者经阑尾、盲肠淋巴结与肠系膜上淋巴结群中的相应淋巴结相联系。盲肠粘膜下层淋巴管内开始出现瓣膜;阑尾淋巴管瓣膜则在阑尾系膜缘内才出现。     

Age-dependent mitogenesis in normal connective tissue cells

We have previously described ways to use the mesentery for studies of proliferation in intact tissue. Here we have studied a weak and a strong mitogenic response in the mesentery of rats aged 6--42 weeks, induced by a single intraperitoneal injection of saline or the mast-cell-degranulating drug 48/80. Proliferation was measured by the specific DNA activity, the fraction of fibroblast- and mesothelial-like cells in the S+G2 cell-cycle-phases, and the mitotic index. We also counted the relative number of mast cells in the tissue (normalized to 5,000 fibroblast-like and mesothelial-like cells), since this might influence 48/80-induced mast-cell-mediated proliferation. In old animals there was a decline in the proliferative response and the time required to initiate DNA synthesis was prolonged. This appears to be the first report of such an age-dependent proliferation characteristic in the mesentery, and probably in any connective tissue. The normalized number of mast cells in the mesentery also declined with increasing age.     

Staining properties of duodenal mast cells in 48/80-treated rats

Summary Mast cells in the tongue, mesentery and lamina propria of the duodenal mucosa in normal and 48/80-treated rats were observed at different time intervals. The tissues were studied comparatively after staining with toluidine blue, acridine orange or alcian bluesafranin. Under the experimental conditions used, the mast cells in the tongue and mesentery showed constant positive reactions to toluidine blue and acridine orange, both of which failed to demonstrate the presence of mast cells in the lamina propria of the duodenal mucosa. The combined alcian blue-safranin stain elicited a safranin-positive reaction in the mast cells of the tongue and mesentery and an alcian blue reaction in those of the lamina propria of the duodenal mucosa. This alcianophilia of the duodenal mast cells was not affected by compound 48/80. On the other hand, the safranin stain of the tongue and mesentery mast cells was altered to alcian blue by the drug. The results are discussed in the light of recent developments in mast cell research.This work was supported by grant MA-2236 of the Medical Research Council of Canada.     

LOCAL MITOGENIC EFFECT OF TISSUE MAST CELL SECRETION

The effect of drug-induced mast cell secretion on proliferation was studied in fibroblast-like and mesothelial-like cells in organ-cultured rat mesentery. Mast cell degranulation achieved by Compound 48/80 was followed by a marked mitogenic reaction in the surrounding tissue cells. The drug itself lacked mitogenic effect on cultured guinea-pig mesentery, the mast cells of which are unresponsive to the drug, and on a human normal fibroblast-like cell line. In contrast, histamine at about 10^?10 M, a major mast cell component, induced marked mitogenesis in guinea-pig mesentery without causing degranulation of mast cells. It is concluded that secreting rat-tissue mast cells release a mitogenic factor or factors acting locally on nearby tissue cells.     

The role of various prostaglandins on the correlation between permeability to albumin and cAMP levels in the isolated mesentery.

Prostaglandins (PG) have been shown to raise the level of cyclic AMP (cAMP) in various tissues, and to increase permeability. Whether both events are linked, is at present a matter of speculation. We have investigated the effects of PGE1, E2, A1, A2, F1alpha and F2alpha on an isolated rat mesentery placed in a diffusion cell (surface area : 2 sq.cm). The PGs (5 microgram/ml) increased the passage of (I 125) - Albumin across the mesentery. In other experiments, diks of rat mesentery (surface area : 2 sq.cm) have been incubated in assay tubes, and cAMP levels measured by a binding protein assay. We have observed an excellent correlation between increases in permeability and cAMP levels (r=0.961). In order of increasing potency on both parameters, the PGs may be classified as follows : PGF, PGA and PGE. In the rat mesentery, under the influence of prostaglandins, increases in permeability and in cAMP levels are apparently connected.     

Dexamethasone blocks the systemic inflammation of alveolar hypoxia at several sites in the inflammatory cascade

Alveolar hypoxia produces a rapid and widespread systemic inflammation in rats. The inflammation is initiated by the release into the circulation of monocyte chemoattractant protein-1 (MCP-1) from alveolar macrophages (AMO) activated by the low alveolar Po(2). Circulating MCP-1 induces mast cell (MC) degranulation with renin release and activation of the local renin-angiotensin system, leading to microvascular leukocyte recruitment and increased vascular permeability. We investigated the effect of dexamethasone, a synthetic anti-inflammatory glucocorticoid, on the development of the systemic inflammation of alveolar hypoxia and its site(s) of action in the inflammatory cascade. The inflammatory steps investigated were the activation of primary cultures of AMO by hypoxia, the degranulation of MCs by MCP-1 in the mesentery microcirculation of rats, and the effect of angiotensin II (ANG II) on the leukocyte/endothelial interface of the mesentery microcirculation. Dexamethasone prevented the mesentery inflammation in conscious rats breathing 10% O(2) for 4 h by acting in all key steps of the inflammatory cascade. Dexamethasone: 1) blocked the hypoxia-induced AMO activation and the release of MCP-1 and abolished the increase in plasma MCP-1 of conscious, hypoxic rats; 2) prevented the MCP-1-induced degranulation of mesentery perivascular MCs and reduced the number of peritoneal MCs, and 3) blocked the leukocyte-endothelial adherence and the extravasation of albumin induced by topical ANG II in the mesentery. The effect at each site was sufficient to prevent the AMO-initiated inflammation of hypoxia. These results may explain the effectiveness of dexamethasone in the treatment of the systemic effects of alveolar hypoxia.     

Effect of Crohn's disease mesenteric mesenchymal stem cells and their extracellular vesicles on T‐cell immunosuppressive capacity

Crohn''s disease (CD) is a chronic inflammatory disease of the gastrointestinal intestinal tract and has characteristic hypertrophic adipose changes observed in the mesentery. To better understand the role of the mesentery in the pathophysiology of Crohn''s disease (CD), we evaluated the immunomodulatory potential of mesenchymal stem cells (MSCs) and their secreted extracellular vesicles (EVs) derived from Crohn''s patients. MSCs and EVs were isolated from the mesentery and subcutaneous tissues of CD patients and healthy individuals subcutaneous tissues, and were analysed for differentiation, cytokine expression, self‐renewal and proliferation. The varying capacity of these tissue‐derived MSCs and EVs to attenuate T‐cell activation was measured in in vitro and an in vivo murine model. RNA sequencing of inflamed Crohn''s disease mesentery tissue revealed an enrichment of T‐cell activation compared to non‐inflamed subcutaneous tissue. MSCs and MSC‐derived EVs isolated from Crohn''s mesentery lose their ability to attenuate DSS‐induced colitis compared to subcutaneous tissue‐derived cell or EV therapy. We found that treatment with subcutaneous isolated MSCs and their EV product compared to Crohn''s mesentery MSCs or EVs, the inhibition of T‐cell proliferation and IFN‐γ, IL‐17a production increased, suggesting a non‐inflamed microenvironment allows for T‐cell inhibition by MSCs/EVs. Our results demonstrate that Crohn''s patient‐derived diseased mesentery tissue MSCs lose their immunosuppressive capacity in the treatment of colitis by distinct regulation of pathogenic T‐cell responses and/or T‐cell infiltration into the colon.     

Cathepsin B containing cells in the rabbit mesentery during invasion of V2 carcinoma cells

Summary Localization of cathepsin B was studied in the rabbit mesentery during invasion of V2 carcinoma cells. Cathepsin B was visualized immunohistochemically by using monospecific sheep antibodies and the avidin-biotinperoxidase complex (ABC) method. Horizontal and vertical semithin Epon embedded sections of stained mesenteries showed that histiocytes always displayed the strongest staining reaction independently of the presence of V2 carcinoma cells. Fibroblasts, mesothelial cells and the invaded V2 cells were less stained. Strongly stained peritoneal monocytes were frequently found on the surface of the mesentery in association with tumor foxi. The role of these various cathepsin B containing cells with respect to extracellular matrix degradation during tumor invasion in the mesentery is not clear; some aspects of this problem are presented in the discussion.     

A morphologic and histochemical study of the mesentery in the guinea pig

The guinea pig mesentery is a uniform, continuous, thin (18 micron) sheet of connective tissue covered by a single layer of flattened mesothelial cells on both surfaces. Tight and gap junctions provide for cell-to-cell adhesion among mesothelial cells. These cells possess numerous micropinocytotic vesicles; a conspicuous basal lamina separates the mesothelium from the underlying connective tissue. Most of the material found between the two serous coverings consisted of a three-dimensional meshwork of abundant collagenous fibers intermingled with a sparse net of very thin (0.4 micron) elastic fibers. Two distinct populations of collagen fibrils are segregated into different compartments of the mesentery. One population is formed of thick (56 nm) fibrils which associate to form closely packed fibers. The second population, composed of loosely arranged thin (38 nm) fibrils which do not become assembled into fibers, is found underlying the basal lamina that separates the mesothelium from the connective tissue. These observations strongly suggest that the mesentery contains both collagens type I and type III. The guinea pig mesentery contains 6.8 mg of sulfated glycosaminoglycans/g dry weight. Most of these glycosaminoglycans (78%) were identified as dermatan sulfate, whilst the rest (22%) corresponded to heparan sulfate.     

Different modes of mesenteric infiltration displayed by two rat leukemias. A study by scanning and transmission electron microscopy and by microcinematography

Infiltration of the mesentery after intraperitoneal implantation of two transplantable rat leukemias, the undifferentiated L5222 and the myeloid BNML, was studied by means of scanning and transmission electron microscopy, and microcinematography. In animals implanted with L5222 cells, contraction of the mesenteric mesothelium is a conspicuous feature. It occurs within the first 24h after implantation and influences decisively the course of infiltration. In contrast, The presence of BNML cells leads to mesothelial contraction only in the terminal stage and, therefore, exerts no direct effect on infiltration. In addition, the two leukemias differ with regard to their cellular motility. Whereas L5222 cells locomote within the mesentery, only stationary movements are recorded with BNML cells. Based on the different interactions with the mesothelium and cell motilities, two distinct modes of infiltrating the mesentery could be ascertained for the two rat leukemias.     

Distribution of extracutaneous melanin pigment in Sparus auratus, Mugil cephalus, and Dicertranchus labrax (Pisces, Teleostei)

The morphological and biochemical characteristics of pigment accumulations found in the kidney, liver, spleen, and mesentery of three different species of teleost fishes have been studied. There are significant differences in number, distribution, and morphology of pigment accumulations in different organs of the three species. Biochemical studies have shown the existence of tyrosinase activity in the mesentery of Mugil cephalus and in the kidney and mesentery of Sparus auratus. No tyrosinase activity was found in any internal organs of Dicertranchus labrax. That activity was assayed using three methods: tyrosine hidroxylation, dopa oxidation, and melanin formation. The morphological and biochemical observations are in agreement. In those organs in which we have demonstrated melanin synthetic activity, the pigment cells are morphologically and like melanophores, while in the organs that show no melanin synthetic activity, the pigment cells resemble macrophages.     

Advanced applications of revascularized free jejunal flaps for difficult wounds of the head and neck

The rationale, technique, and an example of full utilization of the jejunomesenteric free flap have been described as a means of reconstructing complex wounds of the head and neck. Not only jejunum, but also well-vascularized mesentery will protect vascular structures and intestinal anastomoses and, in addition, accept a skin graft. No additional muscular or other flaps are necessary to achieve these ends. Finally, no additional donor-site morbidity is incurred by harvesting the supplementary mesentery.     

The role of various prostaglandins on the correlation between permeability to albumin and cAMP levels in the isolated mesentery

Prostaglandins (PG) have been shown to raise the level of cyclic AMP (cAMP) in various tissues, and to increase permeability. Whether both events are linked, is at present a matter of speculation. We have investigated the effects of PGE1, E2, A1, A2, F1α and F2α on an isolated rat mesentery placed in a diffusion cell (surface area : 2 sq.cm). The PGs (5 μg/ml) increased the passage of (I 125)-Albumin across the mesentery.In other experiments, diks of rat mesentery (surface area : 2 sq.cm) have been incubated in assay tubes, and cAMP levels measured by a binding protein assay. We have observed an excellent correlation between increases in permeability and cAMP levels (r=0.961). In order of increasing potency on both parameters, the PGs may be classified as follows : PGF, PGA and PGE.In the rat mesentery, under the influence of prostaglandins, increases in permeability and in cAMP levels are apparently connected.     

Mast-cell-mediated angiogenesis: a novel experimental model using the rat mesentery

The angiogenic effect of autogenous secreting mast cells (MCs) was studied using a novel experimental approach. The virtually avascular membranous rat mesentery was used as test tissue. The activation of MCs was elicited by repeated intraperitoneal injections of the MC-secretagogue compound 48/80, which per se appears inert from the proliferogenic and angiogenic point of view. Angiogenesis was quantitated histologically and expressed the number of vessels/unit length of mesentery. The smallest vessels recognized had a luminal area of approximately 7-8 microns 2 (corresponding to a circular diameter of 3.0-3.2 microns). Seven to ten days after MC-activation ended, the number of blood vessels had increased 7- to 6-fold. A retrogressive reaction occurred between days 21 and 38 after treatment, when the number of vessels had essentially normalized, as compared to vehicle-treated controls. The present study, introducing the membranous mesentery as a model for quantitative angiogenetic studies, provides evidence that MCs can induce angiogenesis, which is new. The possible therapeutic implication of this finding is noteworthy.     

Development of the innervation of fetal mesenteric microvasculature

Summary The innervation of the mesenteric microvasculature was studied in fetal and neonatal rabbits with the aid of methods demonstrating fluorescence of catecholamines and cholinesterase activity as well as a silver impregnation procedure. The results showed that: (1) adrenergic nerve fibers were present, coursing independently in the mesentery by day twenty-one of gestation, and were found routinely in the adventitia of arterioles and venules by day 25 of gestation; (2) cholinesterase positive cells and fibers of the myenteric plexus were present by day 18 of gestation but cholinergic fibers were not present in the mesentery until day 26; the latter not being associated with blood vessels; and (3) nerve fibers in the mesentery thought to be sensory stained positively with the Holmes silver method on day 18 of gestation.Supported by grants from the Akron Heart Association and the Heart Association of Southwestern Ohio.Recipient of N.I.H. Research Career Development Award AM-42, 370.