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At least 91 records · Page 5Linked to original sources

Stomach-selective gene transfer following the administration of naked plasmid DNA onto the gastric serosal surface in mice.

The purpose of the present study was to achieve a stomach-selective gene transfer following the administration of naked plasmid DNA (pDNA) onto the gastric serosal surface in mice. Gene expression in the stomach and other tissues was evaluated by firefly luciferase activity. Six hours after gastric serosal surface instillation of naked pDNA, high gene expression in the stomach was observed. On the contrary, intravenous and intraperitoneal injection of naked pDNA exhibited no detectable gene expression. Following instillation of naked pDNA onto the gastric serosal surface, gene expression in the stomach was significantly higher than in other tissues. Gene expression in the stomach was highest 12 h after the instillation and thereafter decreased gradually. Utilizing a glass-made diffusion cell that is able to limit the contact dimension between the gastric serosal surface and the naked pDNA solution administered, site-specific gene expression in the stomach was achieved. This novel gene transfer method is expected to be a safe and effective treatment against serious stomach diseases.

Animals↗

De-serosalized muscle layer covering method for antireflux ureteroileostomy: a new operative technique and pressure study with ureterometry at the ureteroileal anastomotic site in dogs.

PURPOSE: In pursuit of a more effective antireflux ureteroileostomy with a lower postoperative complication rate we performed a new operative technique and evaluated intraureteral pressure with ureterometry to examine the mechanism of antireflux function. MATERIALS AND METHODS: A total of 11 beagle dogs were used in this study. A 3 x 2 cm. section of the ileal serosa was removed, the severed ureter was directly anastomosed to the de-serosalized area and 1 cm. of terminal ureter and the direct anastomotic site were covered with the de-serosalized ileal wall. The bladder was augmented with the ileum containing the ureter. Postoperative evaluations were performed monthly and ureterometry of the reimplanted ureter was done 6 months postoperatively. RESULTS: Complete reflux prevention and a low stricture rate were achieved with this procedure. Direct ureteroileal anastomosis caused stricture in 1 of the 11 ureters but the covering procedure to prevent ureteral reflux caused no ureteral strictures. When the bladder was empty, ureteral closure pressure at the intramural portion of the ureter was low. At the phase of high intravesical pressure ureteral closure pressure at the intramural ureter was as high as intravesical pressure. CONCLUSIONS: The de-serosalized muscle layer covering method prevented ureteral reflux completely with a low stricture rate. The antireflux function of this method seems to depend on the flexibility of the terminal ureter covered with the de-serosalized ileal wall. Reflux prevention in the low intravesical pressure phase seems to be due to extension of the ileal wall.

Anastomosis, Surgical↗

Carcinoembryonic antigen mRNA in abdominal cavity as a useful predictor of peritoneal recurrence of gastric cancer with serosal exposure.

Peritoneal dissemination is the most frequent type of recurrence in patients with gastric cancer with serosal exposure, irrespective of whether they have undergone curative gastrectomy. The purpose of this study was to establish a method to detect micrometastatic cells in the abdominal cavity and predict peritoneal recurrence in patients with such gastric carcinomas. A total of 86 patients with gastric carcinoma, undergoing gastrectomy, were examined. Reverse transcriptase-polymerase chain reaction (RT-PCR) assay was used to detect carcinoembryonic antigen (CEA) mRNA in abdominal lavage fluid. Twenty-four cases without serosal exposure were negative, while all 13 cases with macroscopic peritoneal dissemination were positive for CEA mRNA. Among the 49 cases with macroscopic serosal invasion and without peritoneal metastasis, cancer cells were detected in 27 cases with RT-PCR while in only 6 cases with conventional cytology. All cytologically-positive cases were also positive for CEA mRNA. Among the 27 CEA-positive cases, 15 patients (56%) relapsed with peritoneal metastasis within 12 months after gastrectomy. In contrast, none of the 22 CEA-negative cases had peritoneal recurrence within 16-60 months of observation, whereas in 43 cytologically-negative cases, 10 patients relapsed with peritoneal recurrence. As compared with conventional cytological examination, this method would be clinically more beneficial for detecting free cancer cells in the peritoneal cavity and for predicting peritoneal recurrence in gastric carcinoma with serosal invasion.

Abdominal Cavity↗

Intracellular pH (pHi) in gastric surface epithelium is more susceptible to serosal than mucosal acidification.

Intracellular microelectrode techniques were used to examine the effects of mucosal or serosal acidification on intracellular pH (pHi) in gastric surface epithelial cells. Necturus antrum was mounted in a modified Ussing chamber, and pHi was determined from the difference between the potentials recorded by intracellular conventional and pH-sensitive microelectrodes. In tissues bathed with bicarbonate-buffered Ringer's solution (pH 7), acidification of the mucosal solution to pH 4.5 by isotonic replacement of the NaHCO3 with NaCl had no significant effects on pHi. In contrast, acidification of the serosal solution to pH 4.5 by replacing the bicarbonate reduced pHi from 7.32 +/- 0.04 to 6.95 +/- 0.06 (p less than 0.001, n = 8). Similarly, in tissues bathed with HEPES-buffered Ringer's solution (pH 7.0), pHi was unaffected by reducing the mucosal solution pH to 4.5 with HCl but fell 0.21 +/- 0.05 pH units (p less than 0.01, n = 7) during acidification of the serosal solution to pH 6. These results suggest that gastric epithelium is more sensitive to acidification from the serosal than the mucosal side. Such a finding is consistent with the concept of a gastric mucosal barrier to luminal acid. It may also explain the gastric epithelium's greater sensitivity to acute ulceration during systemic acidosis.

Animals↗

[Relationship between free cancer cells in the abdominal cavity, serosal type and pathologic characteristics of gastric cancer].

Of 100 cases of gastric cancer, free cancer cells in the abdominal cavity were detected in 32 (32%). In serosal typing, the free cancer cells were often found in the tendonoid (62.2%) and diffused colour types (60.0%). When the area of serosa invasion was over 20 cm2, the positive rate was 56.6% and only 2.5% if below 20 cm2. Incidence of free cancer cells was related to the depth of cancer infiltration, being often found in S2 and S3. It was also related to the pathologic characteristics of gastric cancer. Free cancer cells were often seen in infiltrating type (Borrmann 3,4); histologically differentiated or undifferentiated adenocarcinomas; nest or diffuse growing types. In patients without metastasis in the peritoneum (P0), the positive rate was 26.1%. This study proved that Chen's serosal classification is correct and useful in assessing whether the cancer cells have penetrated through the serosa during operation. Different treatment should be used in cases with different serosal types. In addition to rational surgical operation, killing of the free cancer cells should be considered in tendonoid and diffused colour serosal types so as to prevent peritoneal metastasis.

Adenocarcinoma↗

Secretory granules of heparin-containing rat serosal mast cells also possess highly sulfated chondroitin sulfate proteoglycans.

Rat serosal mast cells, which synthesize only heparin proteoglycans as detected by intrinsic labeling with [35S]sulfate, were analyzed for the presence of intracellular chondroitin sulfate proteoglycans by chemical and immunochemical means. Rat serosal mast cells of greater than 99% purity were treated with Zwittergent 3-12 and 4 M guanidine HCl, and the extracted nonradiolabeled proteoglycans were purified by density gradient centrifugation. As assessed by quantification of the unsaturated disaccharides released from the proteoglycans by chondroitinase ABC treatment, 10(6) rat serosal mast cells contained 2.4-4.5 micrograms of chondroitin sulfate proteoglycans. Analysis of the chondroitinase ABC digests by high performance liquid chromatography revealed the unsaturated disaccharides delta Di-4S, delta Di-diSB, and delta Di-diSE which were derived from GlcA----GalNAc-4-SO4, iduronic acid-2-SO4----GalNAc-4-SO4, and GlcA----GalNAc-4,6-diSO4, respectively. The molar ratio of the monosulfated to disulfated disaccharides was approximately 2:1 with delta Di-diSE greater than delta Di-diSB. When analyzed with a mouse anti-chondroitin sulfate monoclonal antibody and fluorescein-labeled F(ab')2 goat anti-mouse IgG, approximately 91% of permeabilized and chondroitinase ABC-treated cells in the mast cell preparations exhibited intracellular fluorescence, and the pattern of staining indicated that the chondroitin sulfate molecules were located in the secretory granules. The specificity of the monoclonal antibody for the unsaturated double bond created by chondroitinase ABC treatment of the proteoglycan in situ was established by the absence of fluorescence when the chondroitinase ABC step was omitted or when heparinase digestion was substituted for chondroitinase ABC. Furthermore, the ability of the anti-chondroitin sulfate monoclonal antibody to mediate fluorescence in situ was markedly reduced by absorption with solid-phase chondroitin sulfate proteoglycan that had been chondroitinase ABC-treated, but not by absorption with undigested proteoglycan or with solid-phase heparin. The highly sulfated chondroitin sulfate proteoglycans of rat serosal mast cells are the same type synthesized by the rat mucosal mast cell subclass.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Glucuronidation in isolated perfused rat intestinal segments after mucosal and serosal administration of 1-naphthol.

Glucuronidation of 1-naphthol in the rat intestinal wall was studied in isolated perfused intestinal segments. 1-Naphthol glucuronide appeared on the mucosal as well as on the serosal side of the intestinal segments irrespective of the side (mucosal and/or serosal) on which naphthol was administered. Appearance of naphthol glucuronide was linear with time. A lag phase of 3.9 +/- 0.9 and 4.3 +/- 1.0 min was observed for the appearance on the mucosal and serosal side, respectively. Total glucuronide appearance rate after serosal administration of 10 to 100 microM naphthol was 3- to 4-fold higher than after mucosal administration. It is concluded that glucuronidation after mucosal administration is limited by the entry of naphthol into the metabolizing compartment.

Animals↗

Relationship between area of serosal invasion and intraperitoneal free cancer cells in patients with gastric cancer.

This study, we evaluated the relationship between the macroscopic area of serosal invasion by gastric carcinoma and the incidence of intraperitoneal free cancer cells which were detected by lavage of the Douglas cavity. Between 1976 and 1989, 362 patients with advanced gastric cancer (t3, t4) were gastrectomized and intraperitoneal free cancer cells were recognized in 119 patients (32.9%). Only 17.3% of patients with an area of serosal invasion of less than 10 cm2 had free cancer cells in their peritoneal cavities, however, 68.5% of patients with an area of serosal invasion greater than 20 cm2 had free cancer cells. Even, if curative surgery was performed for patients with free cancer cells, their five-year survival rate was very poor. The five-year survival rate of patients with curatively operated gastric cancer without free cancer cells was 49.3%, however that of patients with free cancer cells was significantly lower (15.4%, P < 0.01). Even when we are not able to receive the cytologic diagnosis for intraperitoneal free cancer cells during the operation, patients with macroscopic areas of serosal invasion greater than 20 cm2 may have free cancer cells in a very high percentage. They should be treated with effective adjuvant chemotherapy the soon after the operation to prevent peritoneal metastatic recurrence and to prolong survival.

Aged↗

Preliminary observations on short-course chemotherapy in tuberculous serositis.

Short-course chemotherapy has not been previously assessed in tuberculous serositis. In this study, 18 patients with tuberculous serositis were studied. 600 mg of rifampicin, 300 mg of isoniazid and 25 mg of pyridoxine were given daily for 6 months. 14 patients completed the therapeutic regimen and were evaluated. 11 of these showed excellent results on the long-term follow-up (18-24 months) and 3 of shorter-term follow-up (9-12 months). Short-course chemotherapy appears to be efficacious in the treatment of tuberculous serositis. However, further and larger trials are recommended.

Adolescent↗

Proposed modifications to 1982 ACR classification criteria for systemic lupus erythematosus: serositis criterion.

The SLICC group believed that the definition of pleuritis should be expanded to include new pleural thickening and pericarditis to include characteristic history. Furthermore, SLICC suggested the addition of abdominal serositis to the current serositis criterion, manifested as either diffuse abdominal pain, with rebound or guarding, and/or ascites or bowel wall edema in the absence of other causes. Abdominal serositis can be secondary to either acute or chronic lupus peritonitis, with the former usually presenting as acute, generalized pain, and the latter as painless ascites.

Humans↗

Enterocytozoon bieneusi as a cause of proliferative serositis in simian immunodeficiency virus-infected immunodeficient macaques (Macaca mulatta).

CONTEXT: Enterocytozoon bieneusi is the most frequent microsporidian parasite of human patients with acquired immunodeficiency syndrome and is a significant cause of diarrhea and wasting. Recently, this organism has also been recognized as a spontaneous infection of several species of captive macaques. As in humans, E bieneusi frequently causes enteropathy and cholangiohepatitis in immunodeficient simian immunodeficiency virus (SIV)-infected macaques. OBJECTIVE: To examine E bieneusi as an etiologic agent of nonsuppurative proliferative serositis in immunodeficient rhesus macaques (Macaca mulatta). DESIGN: Retrospective analysis of necropsy material obtained from immunodeficient SIV-infected rhesus macaques. RESULTS: Examination of SIV-infected rhesus macaques (n = 225) revealed E bieneusi proliferative serositis in 7 of 16 cases of peritonitis of unknown origin. The organism could be identified by in situ hybridization and polymerase chain reaction in sections of pleura and peritoneum obtained at necropsy. Serositis was always accompanied by moderate-to-severe infection of the alimentary tract, and morphologic evidence suggested dissemination through efferent lymphatics. Colabeling experiments revealed most infected cells to be cytokeratin positive and less frequently positive for the macrophage marker CD68. Sequencing of a 607-base pair segment of the small subunit ribosomal gene revealed 100% identity to sequences obtained from rhesus macaques (Genbank accession AF023245) and human patients (Genbank accession AF024657 and L16868). CONCLUSIONS: These findings indicate that E bieneusi disseminates in immunodeficient macaques and may be a cause of peritonitis in the immunocompromised host.

Animals↗

Egg yolk serositis in an American alligator (Alligator mississippiensis).

An adult female American alligator (Alligator mississippiensis) had diffuse, yellow, granular serosal thickening at necropsy. Light microscopic examination of affected stomach, small intestine and spleen revealed a chronic proliferative serositis associated with 3 to 15 micron eosinophilic extracellular globules identified histochemically and morphologically as egg yolk. The intracoelomic egg yolk was considered to be the cause of the serosal reaction.

Alligators and Crocodiles↗

Tumour spread in serosal cavities: what have we learned? Montebello Conference--PSC, Lillehammer, Norway, June 18-22, 2004.

During the Montebello Conference on malignant serosal tumours at Lillehammer, Norway, in June 2004, a group of 30 international experts addressed the biologic and genetic aspects of malignant tumours affecting serosal cavities in the human body. Three neoplasms were mainly dealt with: mesotheliomas arising locally, ovarian carcinomas developing in close proximity to the serosa, and breast tumours in which the spread came from some distance. New, important data on the tumour microenvironment and the process of carcinogenesis with progression and acquisition of invasive properties shed new lights on the mechanisms, including proliferative properties, alterations of signal transduction pathways, and tissue remodelling by proteolytic enzymes in the metastasizing cells. Several of these markers have considerable diagnostic and clinical interest. In addition, new aspects of morphologic and immunocytochemical characteristics of the cells as well as genetic markers may soon become powerful tools for practical use. The molecular fingerprint of the individual tumours may also give guidelines for chemotherapy as well as biologic therapies, including induction of apoptosis. The easy accessibility of tumours from serosal fluids and possibilities for specific discrimination of the neoplastic cells from admixed leukocytes and other cells are promising avenues for cytodiagnostics.

Breast Neoplasms↗

Identification and classification of serosal invasion, as it relates to cancer cell shedding and surgical treatment in gastric cancer.

Surgery to the extent of R+1 [gastric resection plus removal of group 1 lymph nodes (N1) and part of group 2 lymph nodes (N2)] is adequate for cancer confined to the mucosa when the serosa is normal, while operation at the R2 level is adequate for cancer confined to the submucosa (SM) or the muscularis propria (PM) when the serosa is of the reactive type. As the status of serosal involvement advances from normal to reactive, nodular, tendonoid and colour diffused, the lymph node metastatic rate soars and the 5-year survival rate declines in that order. There were no peritoneally shed cancer cells in cases with normal or reactive serosa over the lesion. Shed cancer cells increased significantly when the serosal involvement exceeded 30 cm2. The 2-year survival rate of the patients who had serosal involvement increased by 23.1% when peritoneal lavage with 43 degrees C distilled water was done.

Follow-Up Studies↗

Intestinal thiamin transport in rats. Thiamin and thiamin phosphoester content in the tissue and serosal fluid of everted jejunal sacs.

Rat everted jejunal sacs were incubated at 37 degrees C for 15-60 min in Krebs-Henseleit buffer, pH 7.4, with or without (control experiments) 0.2 microM [thiazole-2-14C]-thiamin. The determination of thiamin and its phosphoesters in the sac wall and in serosal fluid was carried out by an electrophoretic micromethod. Irrespective of the presence of 14C-thiamin, the tissue content of endogenous thiamin pyro- and triphosphate decreased during the incubation, whilst that of thiamin-monophosphate remained relatively constant. The tissue content of free thiamin increased substantially only in control experiments. Endogenous free thiamin, together with a small amount of monophosphate, was found to enter the serosal fluid. The transfer of both compounds was greatly enhanced by the incubation with 14C-thiamin, when an efficient thiamin phosphorylation could be demonstrated. During incubation with 14C-thiamin, the concentration of 14C-thiamin-pyrophosphate and, to a lesser extent, that of free 14C-thiamin increased progressively in the tissue, while 14C- thiamin-monophosphate content remained almost unchanged. No 14C-thiamin-triphosphate was detected. There was a rapid increase in the tissue specific radioactivity of free thiamin and thiamin-monophosphate, which preceded the rise in the specific radioactivity of thiamin-pyrophosphate. The specific radioactivities of the former compounds in the serosal fluid reflected those observed in the intestinal tissue. These results are interpreted as evidence suggesting that the active transport of thiamin is efficient only when intracellular thiamin phosphorylation is operating.

Animals↗

Ca2+- and H+-dependent effects of crude bacterial phospholipase C on the hydroosmotic response of toad urinary bladder to serosal hypertonicity.

Phospholipase C (EC 3.1.4.3.) from Clostridium perfringens (crude extracts) was used to study the role of phospholipids in the osmotic permeability of the urinary bladder of the toad. When added to the serosal bath (430 mU/ml) it inhibited the effects of antidiurectic hormone (ADH) and exogenous cyclic AMP. Under the same conditions the increase in osmotic flow produced by serosal hypertonicity (SH) was slightly enhanced by the lipase. The hydroosmotic effect of SH was greatly potentiated by the lipase by decreasing 10-fold the Ca2+ concentration. The SH-induced flow was inhibited by the lipase if the Ca2+ or the H+ concentration was increased 10-fold, but not if the increase in positive charges was produced by a concentration of Mg2+. Phospholipase C had no effect on the action of either ADH or SH if added to the mucosal bath. Serosal neuraminidase or phospholipase A2 could not mimic the effect of phospholipase C on SH. The effect of phospholipase C on the response to SH was not modified if fatty acid-free bovine serum albumin was added to the bath. Therefore, the release of products of lipolysis into the bath do not seem to be responsible for the effects of phospholipase C on SH-induced water flow. The results suggest that the effects of the enzyme on the composition and rearrangement of lipids at the basolateral membrane produce modifications of the water flow. Ca2+ and H+ may modify the enzyme-substrate interaction, suggesting that different phospholipids may be differentially involved in the control of water permeability of the basolateral membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Methyl-3-isobutylxanthine↗

Fecal peritonitis: microbial adherence to serosal mesothelium and resistance to peritoneal lavage.

Fecal contamination of the peritoneal cavity is a serious and potentially life-threatening event. While numerous models have been developed to study the pathogenesis of intraabdominal infection, to date, most investigations have failed to focus on the adherence of the contaminants to the serosal mesothelium. In the present investigation, the cecal ligation and puncture technique (CLP) was performed in Sprague-Dawley rats to study the following: (a) the kinetics of microbial adherence to the serosal mesothelium, (b) the stability of the aerobic and anaerobic intraperitoneal/mesothelial populations, following extended saline lavage, and (c) the impact of antimicrobial lavage on the stability of the mesothelial microbial populations. The Enterobacteriaceae rapidly colonized the serosal mesothelium and were the predominant flora up to 4 hours post-CLP. After 8 hours, the Bacteroides fragilis group represented the predominant peritoneal wash and mesothelial-associated microorganisms. Extended saline lavage failed to significantly reduce the mesothelial microbial populations. While antimicrobial lavage produced an immediate decrease in mesothelial microbial recovery, the results were transitory and the microbial populations achieved or exceeded prelavage levels at 24 hours postlavage. Microbial colonization of the peritoneal mesothelial surface is a rapid and stable phenomena following penetrating injury to the distal bowel. The results further suggest that the mesothelial populations are resistant to intraperitoneal lavage.

Animals↗

Basolateral membrane potential and conductance in frog skin exposed to high serosal potassium.

In studies of apical membrane current-voltage relationships, in order to avoid laborious intracellular microelectrode techniques, tight epithelia are commonly exposed to high serosal K concentrations. This approach depends on the assumptions that high serosal K reduces the basolateral membrane resistance and potential to insignificantly low levels, so that transepithelial values can be attributed to the apical membrane. We have here examined the validity of these assumptions in frog skins (Rana pipiens pipiens). The skins were equilibrated in NaCl Ringer's solutions, with transepithelial voltage Vt clamped (except for brief perturbations delta Vt) at zero. The skins were impaled from the outer surface with 1.5 M KCl-filled microelectrodes (Rel greater than 30 M omega). The transepithelial (short-circuit) current It and conductance gt = -delta It/delta Vt, the outer membrane voltage Vo (apical reference) and voltage-divider ratio (Fo = delta Vo/delta Vt), and the microelectrode resistance Rel were recorded continuously. Intermittent brief apical exposure to 20 microM amiloride permitted estimation of cellular (c) and paracellular (p) currents and conductances. The basolateral (inner) membrane conductance was estimated by two independent means: either from values of gt and Fo before and after amiloride or as the ratio of changes (-delta Ic/delta Vi) induced by amiloride. On serosal substitution of Na by K, within about 10 min, Ic declined and gt increased markedly, mainly as a consequence of increase in gp. The basolateral membrane voltage Vi (= -Vo) was depolarized from 75 +/- 4 to 2 +/- 1 mV [mean +/- SEM (n = 6)], and was partially repolarized following amiloride to 5 +/- 2 mV.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗