Smoking increases the incidence of complicated diverticular disease of the sigmoid colon.

BACKGROUND AND AIMS: The aim of this study was to establish whether smoking is associated with complicated diverticular disease and adverse outcomes of operative treatment of diverticular disease. Smoking has been associated with increased rate of perforations in acute appendicitis as well as failure of colonic anastomosis in patients resected for colonic tumours. It has also been suggested that smoking is a risk factor for complicated diverticular disease of the colon. MATERIAL AND METHODS: Retrospective investigation of records of 261 patients electively operated for diverticular disease in Helsinki University Central Hospital during a period of five years. RESULTS: The smokers underwent sigmoidectomy at a younger age than the non-smokers (p = 0.001) and they had an increased rate of perforations (p = 0.040) and postoperative recurrent diverticulitis episodes (p = 0.019). CONCLUSIONS: We conclude that smoking increases the likelihood of complications in diverticulosis coli. The development of complicated disease also seems to proceed more rapidly in smokers.Key words: Sigmoid resection; laparoscopy; laparoscopic sigmoidectomy; smoking and diverticular disease; complicated diverticular disease; diverticulitis.

Is routine dilatation after repair of esophageal atresia with distal fistula better than dilatation when symptoms arise? Comparison of results of two European pediatric surgical centers.

BACKGROUND/PURPOSE: The aim of this study was to determine whether routine dilatation of the anastomosis after repair of an esophageal atresia with distal fistula (EADF) is superior to a wait-and-see policy with dilatation only when symptoms arise. METHODS: The records of 100 consecutive patients operated on for EADF in 2 European pediatric surgical centers (A [n = 63], B [n = 37]) were reviewed. In center A, dilatation of the anastomosis was carried out in symptomatic cases only, whereas in center B dilatation was begun 3 weeks postoperatively and repeated every 1-3 weeks until a stable diameter of 10 mm was reached. Particular attention was paid to the number of dilatations per patient, dilatation-related complications, and differences in results after 2 years. RESULTS: The patient materials of both centers did not differ with respect to the incidence of prematurity, tracheomalacia, gastroesophageal reflux (GER), and major postoperative complications. The incidence of associated anomalies was higher in center B (P < .05). In center A, 26 of 63 patients underwent dilatation; in center B, all 37 patients were dilated (P < .05). Median number of dilatations per patient was 4 in center A and 7 in center B (P < .05). In center A, 23 of 26 and in center B, 20 of 37 of the patients received medical treatment for GER at the time of the dilatations. Dilatation-related complications developed in 7 of 26 patients of center A and in 3 of 37 patients in the center B (P value, not significant). The median primary hospital stay was 24 days in center A and 33 days in center B (P < .05), median secondary hospital stay for dilatation was 6 days in center A and 13 days in center B (P < .05). After 2 years of follow-up, the incidence of dysphagia, respiratory problems, or bolus obstruction did not differ significantly between the 2 centers. CONCLUSIONS: A wait-and-see policy and dilatations based on clinical indications for patients with repaired EADF is superior to routine dilatations. It appears that more than half of the patients do not require dilatations at all.

EC-SOD gene therapy reduces paracetamol-induced liver damage in mice.

BACKGROUND: Paracetamol overdose causes acute liver damage which leads to severe centrilobular hepatic necrosis. The hepatotoxic effect is caused by reactive metabolites and oxidative stress. Since extracellular superoxide dismutase (EC-SOD) protects tissues against the harmful effects of superoxide anion, the hypothesis that systemic adenovirus-mediated EC-SOD gene transfer could reduce liver damage was tested. METHODS: Mice were given paracetamol (600 mg/kg) enterally 2 days after adenovirus-mediated gene transfer of EC-SOD (2 x 10(9) pfu). Five days after gene transfer, plasma and tissue samples were collected for clinical chemistry analyses and tissue pathology evaluation. RESULTS: EC-SOD was expressed in a dose-dependent manner with the highest enzyme activity occurring 3 days after the gene transfer. Clinical chemistry and tissue pathology analyses showed that adenoviral EC-SOD gene transfer significantly attenuated release of liver enzymes and inhibited necrosis and apoptosis caused by paracetamol overdose. CONCLUSION: The results indicate the involvement of superoxide anion in paracetamol-mediated liver damage and suggest a possible protective role for EC-SOD gene transfer in paracetamol-induced liver damage.

Gene transfer of extracellular superoxide dismutase to atherosclerotic mice.

Clinical and epidemiological studies have provided circumstantial evidence that oxidized low-density lipoprotein (LDL) and antioxidants are involved in the pathogenesis of atherosclerosis. Superoxide dismutases (SODs) have been shown in vitro to protect LDL from deleterious effects of superoxide anions. In the present study, we have used adenoviral gene transfer to determine effect of extracellular SOD (EC-SOD) on atherogenesis in LDL receptor -/- mice. Intravenous administration of EC-SOD adenovirus (2 x 10(9) plaque forming units) into tail vein targeted transgene mainly to liver and induced a 3.5- to sevenfold increase in plasma total SOD activity. EC-SOD was secreted into circulation for 2-3 weeks mostly in a truncated B-form, suggesting that endogenous proteolytic mechanisms control the level and distribution of the enzyme. Therapeutic potential was determined by measuring plasma resistance against copper oxidation and analyzing atherosclerotic lesion areas in aortas of LDL receptor -/- mice. Mice were kept on a cholesterol diet for 10 weeks before gene transfer and 3 or 6 weeks after the gene transfer. Results showed a tendency for a reduction in the overall lesion area after EC-SOD gene transfer as compared with LacZ transduced control mice, but the difference did not reach statistical significance. It is concluded that short-term overexpression of EC-SOD in vivo does not affect atherogenesis in LDL receptor -/- mice.

Rabbit extracellular superoxide dismutase: expression and effect on LDL oxidation.

Extracellular superoxide dismutase (EC-SOD) is a secreted antioxidative enzyme with an abundant mRNA expression in kidney and arterial wall. In order to study expression and antioxidative function of EC-SOD, we cloned the rabbit ec-sod cDNA and produced the recombinant protein in cell culture. In vitro studies did not show a direct relationship between the amounts of synthesized mRNA and secreted protein activity, suggesting post-transcriptional regulation. The antiatherogenic role of EC-SOD was studied by determining the effect of EC-SOD on the oxidation (ox) of low density lipoprotein (LDL), and subsequent degradation of oxLDL in RAW 264 macrophages in vitro. It was found that recombinant EC-SOD reduced both the degradation of LDL in RAW 264 macrophages by 28-36% and its electrophoretic mobility caused by endothelial cell-mediated oxidation. It is therefore suggested that EC-SOD can act as a protective enzyme against the development of atherosclerosis.

Hirudin stimulates prostacyclin but not endothelin-1 production in cultured human vascular endothelial cells.

To study the effect of hirudin on endothelial cell prostacyclin (PGI2) and endothelin-1 (ET-1) production, we cultured human umbilical vein endothelial cells (HUVECs), stimulated them with 0.00001-10 kU/l of hirudin for 12-24 hours, and measured by radioimmunoassays the concentrations of 6-ketoprostaglandinF1 alfa (6-keto, a metabolite of PGI2) and ET-1 in the incubation medium. In incubation medium containing 10% serum hirudin stimulated PGI2-production dose-dependently. The lowest stimulatory hirudin concentration was 0.001 kU/l, which increased the concentration of 6-keto by 10.8 +/- 4.4% (mean +/- S.E) (p < 0.01). The greatest stimulation rate (28.6 +/- 6.2%, p < 0.001) was obtained with the highest hirudin concentration (10 kU/l), when the culture medium contained 10% human serum. The PGI2-stimulating activity was exaggerated in the absence of serum, when 1 kU/l of hirudin increased PGI2-production by 59.7 +/- 6.2% (p < 0.001, n = 14). Stimulation of PGI2 appeared after 12 hour incubation. Hirudin had no effect on the conversion of exogenous arachidonic acid to 6-keto or on the production of ET-1. We thus conclude that hirudin stimulates PGI2-production through de novo protein synthesis. Stimulation of PGI2-production by hirudin may contribute to its antithrombotic activity, since PGI2 favours vasodilatation and attenuates platelet aggregation.

17 beta-estradiol stimulates prostacyclin, but not endothelin-1, production in human vascular endothelial cells.

The exact mechanisms by which estrogens protect against occlusive vascular disorders are not known. One possibility could be an effect on vascular endothelial vasoactive compounds, such as vasodilatory prostacyclin (PGI2) and vasoconstrictory endothelin (ET-1). Here we report on the effect of 17 beta-estradiol on the synthesis of PGI2 and ET-1 in cultured human umbilical vein endothelial cells. These cells were incubated in the absence (control) and presence of 17 beta-estradiol (0.001-1 mumol/L) for 3-24 h with serum (10%) or without serum. The release of PGI2, as assessed by its metabolite 6-keto-prostaglandin F1 alpha, and that of ET-1, were assessed by RIA. 17 beta-Estradiol (0.01-0.1 mumol/L) predissolved in ethanol (final concentration, 0.01%) increased PGI2 production by 26-30% in endothelial cells incubated without serum. This increase in PGI2 production was enhanced up to 66% when 17 beta-estradiol (1 mumol/L) was encapsulated within beta-cyclodextrin. The stimulation of PGI2 production was detectable after 12 h of incubation. The 17 beta-estradiol-induced stimulation of PGI2 production was blocked in dose-dependent manner by antiestrogenic tamoxifen. 17 beta-Estradiol failed to affect the production of PGI2 if the endothelial cells were incubated with serum and had no effect on ET-1 production under any conditions. 17 beta-Estradiol-induced stimulation of vasodilatory and antiaggregatory PGI2 production without a concomitant change in vasoconstrictory ET-1 production may provide one explanation for the ability of estradiol to maintain vascular health and protect against vascular disorders.

New sensitive and specific assay for human immunodeficiency virus antibodies using labeled recombinant fusion protein and time-resolved fluoroimmunoassay.

A new, rapid method for the detection of human immunodeficiency virus type 1 (HIV-1) antibody by time-resolved fluoroimmunoassay (TR-FIA) was developed. In this assay format, microtitration strips were coated with a recombinant fusion protein, and the same protein was labeled with europium and added into the wells simultaneously with the test specimens. The recombinant fusion protein contained the HIV-1 p24 gag protein sequence that carried an insertion, near the carboxyl terminus, of a 23-amino-acid sequence from a highly conserved region of the HIV-1 gp41 envelope protein. This recombinant antigen enabled the detection of antibodies to both gag and env gene products. When this assay was compared with a commercially available recombinant enzyme-linked immunoabsorbent assay (ELISA) by using four quality-control panels, the TR-FIA detected all 20 positive specimens, while the recombinant ELISA detected only 16 of them. This increased sensitivity could be demonstrated directly by the assay of dilution series of HIV-1-positive sera. The analysis of two seroconversion panels by TR-FIA and six ELISAs showed that TR-FIA allowed detection of antibody in infected individuals 16 days earlier than the other assays did. In addition to being highly sensitive, the assay was highly specific; of the 57 samples shown to be repeatedly positive by ELISA but known to be HIV-1 negative by Western immunoblot analysis, only 1 sample reacted positively in this assay. The specificity of the assay was 99.9% when 1.054 random serum specimens were tested.

Group-specific component (Gc): subtypes in the Finnish population. Description of a new allele and an apparent mother-child incompatibility.

The group-specific component (Gc) subtypes were determined in 575 adult Finns by immunoblotting after isoelectric focusing in agarose gel. The gene frequencies were Gc1S = 0.661, Gc1F = 0.139 and Gc2 = 0.200. This material included one rare allele, a more acidically focusing Gc 2 (named Gc 2A18). The phenotypes of 200 mother-child pairs studied were in accordance with the three-allelic mode of inheritance. An apparent mother-child incompatibility observed during routine paternity testing is reported.

Determination of cis,cis-methylene interrupted polyunsaturated fatty acids in aqueous solutions by lipoxygenase chemiluminescence.

The chemiluminescent reaction of luminol during lipoxygenase-catalyzed oxygenations was studied with the purpose of developing a specific luminometric assay for cis,cis-1,4-pentadiene fatty acids directly in aqueous solutions. The addition of picomole levels of either linoleic or arachidonic acids to reaction systems containing 0.04 mM luminol and 40 micrograms/ml of purified soybean lipoxygenase-1 gave light emission curves with a single sharp maximum. Under these conditions the peak heights were linearly dependent on the fatty acid concentration and the detection limit for both of the fatty acids was 2 pmol with a signal to noise ratio of 2. For maximum reproducibility of the assays a procedure for the proper quantitation of the enzyme was developed. The fact that the assay proved to be relatively interference-free was ascribed to the high molar enzyme/substrate ratio (above 1).