Tezosentan normalizes hepatomesenteric perfusion in a porcine model of cardiac tamponade.

BACKGROUND: To investigate endothelin-1 (ET-1)-dependent hepatic and mesenteric vasoconstriction, and oxygen and lactate fluxes in an acute, fixed low cardiac output (CO) state. METHODS: Sixteen anesthetized, mechanically ventilated pigs were studied. Cardiac tamponade was established to reduce portal venous blood flow (Q(PV)) to 2/3 of the baseline value. CO, hepatic artery blood flow (Q(HA)), Q(PV), hepatic laser-Doppler flow (LDF), hepatic venous and portal pressure, and hepatic and mesenteric oxygen and lactate fluxes were measured. Hepatic arterial (R(HA)), portal (R(HP)) and mesenteric (R(mes)) vascular resistances were calculated. The combined ET(A)-ET(B) receptor antagonist tezosentan (RO 61-0612) or normal saline vehicle was infused in the low CO state. Measurements were made at baseline, after 30, 60, 90 min of tamponade, and 30, 60, 90 min following the infusion of tesozentan at 1 mg/kg/h. RESULTS: Tamponade decreased CO, Q(PV), Q(HA), LDF, hepatic and mesenteric oxygen delivery, while hepatic and mesenteric oxygen extraction and lactate release increased. R(HA), R(HP) and R(mes) all increased. Ninety minutes after tesozentan, Q(PV), LDF and hepatic and mesenteric oxygen delivery and extraction increased approaching baseline values, but no effect was seen on CO or Q(HA). Hepatic and mesenteric handling of lactate converted to extraction. R(HA), R(HP) and R(mes) returned to baseline values. No changes were observed in these variables among control animals not receiving tesozentan. CONCLUSION: In a porcine model of acute splanchnic hypoperfusion, unselective ET-1 blockade restored hepatomesenteric perfusion and reversed lactate metabolism. These observations might be relevant when considering liver protection in low CO states.

Inappropriate high-rate ventricular pacing in a patient with a defibrillator.

We describe a patient who presented with high rate ventricular pacing secondary to dysfunction of his implantable cardioverter defibrillator (ICD). The device was also unable to communicate with the programmer and unable to treat ventricular fibrillation. Immediate disconnection of the ICD from the leads was the only effective recourse. Subsequent detailed technical analysis of the device revealed a different electrical circuit problem from that found in typical cases of runaway pacing. To our knowledge this is the first description of a malfunction of precisely this nature.

Endothelin-1 blockade corrects mesenteric hypoperfusion in a porcine low cardiac output model.

OBJECTIVE: To study the importance of endothelin-1-induced vasoconstriction in a model of acute and maintained low cardiac output, by investigating regional changes within the mesenteric and particularly the intestinal mucosal circulation. DESIGN: Prospective, controlled animal study. SETTING: University-affiliated research laboratory. SUBJECTS: Thirteen fasted, anesthetized, mechanically ventilated landrace pigs. MEASUREMENTS AND MAIN RESULTS: Cardiac output, portal venous blood flow, renal arterial flow, jejunal mucosal microcirculation by laser Doppler flowmetry, jejunal capnotonometry (Pco2 gap), and jejunal mucosal oxygenation (tPo2) were monitored. Cardiac tamponade was established to reduce portal venous blood flow to a preset end point at two thirds of baseline. Measurements were made at baseline, after 90 mins of cardiac tamponade, and 90 mins after the administration of the combined endothelinA/endothelinB antagonist tezosentan at 1 mg.kg-1.hr-1 during tamponade in seven animals. Six animals served as time controls and received only the vehicle. Cardiac tamponade decreased portal venous blood flow, renal arterial flow, and laser Doppler flowmetry, whereas the Pco2 gap increased. The change in tPo2 failed to gain statistical significance (p =.08). Administration of tezosentan during tamponade restored portal venous blood flow and laser Doppler flowmetry to baseline values, increased tPo2 above baseline, and decreased Pco2 gap. No effect on renal arterial flow was observed. Investigated variables remained unchanged in control animals after induction of cardiac tamponade. CONCLUSIONS: Endothelin-1 blockade in acute cardiac failure improves mesenteric, but not renal, perfusion, illustrating the regional importance of endothelin-1-induced vasoconstriction. Importantly, endothelin-1 blockade restored mucosal blood flow and oxygenation, which might be particularly interesting considering the implications for maintenance of mucosal barrier integrity in low output states.

Aluminum potentiates P(i) transport stimulation induced by fluoride in osteoblast-like cells.

The effect of aluminum (AI) on inorganic phosphate (P(i)) transport stimulation induced by fluoride (F) was investigated in MC3T3-E1 osteoblast-like cells. Al potentiated the increase in P(i) transport activity induced by F in a dose- and time-dependent manner. Results obtained with deferoxamine mesylate, an Al chelator, suggest that a fluoroalumino complex is probably the active F molecule responsible for the change in P(i) transport observed in this study. The signaling pathway responsible for the stimulation of P(i) transport by F+Al likely involves a tyrosine phosphorylation process but neither a protein kinase C nor a mitogen-activated protein kinase pathway. As previously found in UMR-106 cells for F alone, F+Al potentiated the change in P(i) transport induced by fetal calf serum. A similar interaction was found between F+Al and thrombin acting through a G protein-coupled receptor. These observations are compatible with the hypothesis that F+Al could interact with G protein-coupled receptors associated with a signaling tyrosine phosphorylation process involved in the regulation of P(i), transport in osteoblast-like cells.

Aluminum potentiates the effect of fluoride on tyrosine phosphorylation and osteoblast replication in vitro and bone mass in vivo.

Osteosclerosis in workers exposed to fluoride (F) and aluminum (Al) (industrial fluorosis) led to the use of F as a treatment to increase bone mass in osteoporosis patients. Because the influence of traces of Al on the effects of F on bone formation is heretofore unknown, we have investigated this issue both in vitro and in vivo. We have found that minute amounts of Al (< or = 10(-5) M) potentiate the effects of F in vitro such that osteoblast proliferation increased by 15 +/- 2.7% at 50 microM (p < 0.001) and by 117.6 +/- 5.1% at 750 microM (p < 0.001), concentrations of F with no mitogenic effect alone. F + Al time-dependently modulated a growth factor signaling pathway(s) associated with enhanced tyrosine phosphorylation (TyrP) of several proteins (p90 [2.9x], p77 [4.9x], p68 [9.6x], and mitogen activated protein kinases [3x]). TyrP was only slightly or not at all changed by F and Al alone, respectively. The effects of F + Al on TyrP and cell proliferation were markedly reduced by 100 microM tyrphostin-51, a tyrosine kinase inhibitor. Protein kinase A (PKA) and protein kinase C (PKC) pathways were not involved in this response. In vivo, F + Al administered for 8 months, at doses that had no effect when the minerals were administered individually, significantly enhanced proximal tibia bone mineral density (BMD) by 6.3 +/- 1% compared with initial values and by 2-fold compared with control ovariectomized rats (p < 0.0001). These effects are consistent with a crucial role of Al in osteosclerosis observed in industrial fluorosis. The results suggest that the combination of F + Al modulates a growth factor-dependent TyrP pathway enhancing mitogen-activated protein kinase and osteoblastic proliferation and bone mass.

Fluoride increases tyrosine kinase activity in osteoblast-like cells: regulatory role for the stimulation of cell proliferation and Pi transport across the plasma membrane.

Fluoride is one of the most effective agents for the treatment of vertebral osteoporosis because of its ability to increase osteoblast proliferation. The present study further investigates the role of protein tyrosine phosphorylation previously suggested to mediate the mitogenic effect of fluoride on bone-forming cells. The activity of the plasma membrane Na-coupled Pi transport system was monitored to assess the relationship between alterations in tyrosine phosphorylation and osteoblast activity induced by fluoride. The results indicate that vanadate, a selective inhibitor of tyrosine phosphatase, mimicked the stimulatory effect of fluoride on Pi transport. The change in Pi transport induced by fluoride was dose dependently inhibited by genistein, a potent inhibitor of tyrosine kinase. Genistein also inhibited the change in cell proliferation induced by fluoride. Associated with these observations, tyrosine phosphorylation activity was significantly increased in subcellular fractions isolated from UMR-106 cells treated with fluoride as compared with those isolated from vehicle-treated cells. This change in tyrosine phosphorylation activity was markedly blunted when genistein was added to the kinase assay buffer. It was not associated with any alteration in specific tyrosine phosphatase activity. There was also no evidence of a direct effect of fluoride on tyrosine phosphatase activity in isolated plasma membrane of UMR-106 cells. In conclusion, the results of the present study suggest that fluoride enhances protein tyrosine phosphorylation in osteoblast-like cells by enhancing tyrosine kinase activity. The results further support the hypothesis that this signal transduction mechanism is involved in the osteogenic effects of fluoride.

Glycoprotein-specific immune responses in cats after exposure to feline herpesvirus-1.

To obtain synchronous infection, 10 cats were inoculated with feline herpesvirus-1 (FHV-1) on the oral, nasal, and conjunctival mucosa. Swab specimens of the nasal, conjunctival, and pharyngeal mucosa were obtained for virus isolation from each cat before inoculation and at 3-day intervals thereafter until postinoculation day 21. Recovery of virus and evidence of clinical signs were used to document FHV-1 infection. Serum was obtained from blood samples collected sequentially from each cat between day 0 and postinoculation day 90. Virus-neutralizing antibody titer was determined in all serum specimens. Immunoprecipitation with [35S]methionine- and [14C]glucosamine-labeled viral antigens, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was performed on each specimen. Three precipitation bands with approximate molecular weights of 105,000, 68,000, and 60,000 were separated from [14C]glucosamine- and [35S]methionine-labeled immunoprecipitates. The concurrent detection of virus-neutralizing antibody and glycoprotein-specific immunoprecipitins implied that in cats, the FHV-1 glycoproteins were important in the induction of virus-neutralizing antibodies to FHV-1.

Bile duct obstruction secondary to chronic pancreatitis in seven dogs.

Seven icteric dogs were determined to have bile duct obstruction secondary to chronic pancreatitis. All dogs had histories of intermittent vomiting and diarrhea. Alkaline phosphatase and alanine aminotransferase activities and total bilirubin concentrations were markedly elevated. Diagnosis was based on exploratory laparotomy and histological examination. Each dog had a 3 to 10 cm mass in the body of the pancreas and obstruction of the common bile duct. Three dogs treated with pancreatectomy, gastrojejunostomy, and cholecystojejunostomy died within five weeks. Three dogs treated with conservative surgical procedures were alive at 8, 16, and 26 months postoperatively. One dog was euthanized because of suspected neoplasia. Hepatic enzyme activity and bilirubin levels decreased markedly in the surviving dogs. Histological examination of the pancreatic masses indicated chronic pancreatitis. Hepatic biopsies revealed evidence of cholestasis. Chronic pancreatitis should be included in the differential diagnoses of icterus, bile duct obstruction, and masses in the pancreas.

Enumeration of total coliforms, fecal coliforms, and Escherichia coli in foods by hydrophobic grid membrane filter: collaborative study.

A collaborative study was conducted in 18 laboratories to assess the performance of the hydrophobic grid membrane filter method against that of the AOAC official first action method 46.013-46.016 for enumerating total and fecal coliforms and Escherichia coli. The study was carried out on frozen breaded fish, raw comminuted poultry, unroasted walnut pieces, ground black pepper, and cheddar cheese. The hydrophobic grid membrane filter method recovered significantly larger numbers of target bacteria in 7 of the food/analysis combinations: fecal coliforms in fish; E. coli in poultry; fecal coliforms and E. coli in walnuts; and total coliforms, fecal coliforms and E. coli in black pepper. Random error (Sr2) associated with the hydrophobic grid membrane filter method was significantly lower than that of the reference method in over 30% of the paired sample series. The hydrophobic grid membrane filter method for total coliform, fecal coliform, and E. coli enumeration in foods has been adopted official first action.

Collaborative study of the MPN, Anderson-Baird-Parker direct plating, and hydrophobic grid-membrane filter methods for the enumeration of Escherichia coli biotype I in foods.

Five Health Protection Branch laboratories compared two membrane filter methods (the Anderson-Baird-Parker direct plating, and a hydrophobic grid-membrane filter method) against the most probable number procedure (MPN) for enumerating Escherichia coli biotype I in foods. Results were available in 24 h by both membrane filter methods, compared with 10-14 days by the MPN procedure. For ground beef, Parmesan cheese, and cut green beans, the hydrophobic grid method generally gave the highest recovery, although the two membrane filter methods were not significantly different. Both these methods gave significantly higher recoveries than the MPN procedure, and for most foods, either method would be preferable. Further work is required before either membrane filter method can be recommended for bean and alfalfa sprouts, which may contain very high levels of Klebsiella spp.

Detection of Salmonella in Refrigerated Preenrichment and Enrichment Broth Cultures.

Refrigeration (4 C) of non-selective and selective enrichment broth cultures for 72 h did not markedly affect detection of Salmonella in 160 contaminated high and low moisture foods. Detection in refrigerated preenrichment (non-selective) broth cultures of poultry and high and low moisture foods was 90, 95 and 100%, respectively; homologous results for refrigerated selective enrichment broth cultures were 90, 100 and 100%. All but one of the 22 negative results were obtained with poultry and two of the six laboratories participating in poultry analysis contributed 19 of the 21 negative results. Refrigeration of broth cultures provides greater operational flexibility by increasing the number of days on which analyses can be initiated without engendering work outside a normal work week.

Microbiological Standards for Cheese: Survey and Viewpoint of the Canadian Health Protection Branch.

The Health Protection Branch, Health and Welfare Canada is considering proposals for microbiological standards for cheese. These proposals are based on a 2-year study (1974-1976) carried out by the Branch. The proposed standards per gram are: total coliforms m = 500, M = 1500, fecal coliforms m = 100, M = 500, and Staphylococcus aureus m = 100, M = 1000, for cheeses made from pasteurized milk; total coliforms m = 5000, M = 50,000, fecal coliforms m = 500, M = 1000, and S. aureus m = 1,000, M = 10,000 for cheeses made from heat treated or unpasteurized milk. The type of standard proposed will be based on a three-class acceptance plan as developed by the International Commission on Microbiological Specifications for Foods. Use of this plan in interpretation of the analytical results allows for the normal variation between analytical samples.