Environment and T regulatory cells in allergy.

The central role of T regulatory cells in the responses against harmless environmental antigens has been confirmed by many studies. Impaired T regulatory cell function is implicated in many pathological conditions, particularly allergic diseases. The "hygiene hypothesis" suggests that infections and infestations may play a protective role for allergy, whereas environmental pollutants favor the development of allergic diseases. Developing countries suffer from a variety of infections and are also facing an increasing diffusion of environmental pollutants. In these countries allergies increase in relation to the spreading use of xenobiotics (pesticides, herbicides, pollution, etc.) with a rate similar to those of developed countries, overcoming the protective effects of infections. We review here the main mechanisms of non-self tolerance, with particular regard to relations between T regulatory cell activity, infections and infestations such as helminthiasis, and exposure to environmental xenobiotics with relevant diffusion in developing countries.

Cardiac surgery for octogenarians--a suitable procedure? Twelve-year operative and post-hospital mortality in 641 patients over 80 years of age.

OBJECTIVES: The increase in life expectancy as a result of therapeutic improvements subsequently leads to a large number of patients with advanced age. The aim of this study was to review the 30-day mortality and mid-term outcome of octogenarians undergoing coronary artery bypass grafting (CABG) or valve replacement (AVR/MVR). METHODS: The data of 641 patients with a mean age of 82.6 years (range 80.0 - 92.6), operated between 9/93 and 12/05, were reviewed. 432 patients underwent CABG, 188 had AVR and 21 had MVR. We analysed peri-/postoperative mortality and clinical outcomes. Follow-up was obtained by phone contact with patients or their physician. Mid-term survival was determined for the whole population by the Kaplan-Meier method; peri- and postoperative risk factor analysis was done using logistic regression. Follow-up ranged from 0.1 to 11.8 years (mean 3.6 +/- 2.6) and was complete for 99%. RESULTS: We observed a perioperative mortality of 8.8% for CABG, 4.8% for AVR and 9.5% for MVR. Perioperative mortality was strongly associated with urgent/emergent operations (P < 0.03), poorer clinical status (P < 0.03), renal dysfunction (P < 0.05) and male gender (P < 0.04). Actuarial survival after 3, 5 and 8 years was as follows: CABG 78%, 66% and 44%; AVR 79%, 68% and 38%; MVR 76%, 61% and 23%. The mean NYHA functional class for survivors improved in the group of patients with CABG from 2.7 to 2.0 (P < 0.03), in the AVR group from 2.8 to 2.0 (P < 0.03), and in the MVR group from 2.9 to 2.3 (P < 0.05). More than 80% of all surviving patients live at home, either alone or with their family. CONCLUSION: In our cohort of octogenarians, cardiac surgery was found to be associated with an acceptable, although increased perioperative mortality. Despite the enhanced perioperative risk, the clinical benefit, as verified by improved functional status and satisfactory mid-term survival rates, justifies surgery in these patients with advanced age.

Is extreme obesity a risk factor for increased in-hospital mortality and postoperative morbidity after cardiac surgery? Results of 2251 obese patients with BMI of 30 to 50.

OBJECTIVE: The number of patients with extreme obesity requiring cardiac surgery is increasing. The aim of the present study was to evaluate the perioperative outcome, complication rate and 30-day mortality of these patients. METHODS: Data of 13 139 patients (9584 males, 3555 females) undergoing cardiac surgery from 1/94 to 12/04 were analyzed. We compared 2251 obese patients with a BMI of 30 to 50 (mean age 64.3 years; group A) with 10 888 non-obese patients (mean age 66.1 years; group B). The incidence of diabetes mellitus was 34.5 % in group A, and 24.2 % in group B. RESULTS: Isolated CABG was performed in 1771 patients from group A (3.4 grafts/pt.) and in 8066 patients from group B (3.2 grafts/pt.). Concomitant procedures were performed in 460 patients (group A) vs. 2645 (group B). Frequency of bilateral internal thoracic artery grafting was 49.4 % (group A) vs. 44.9 % (group B). There were no significant differences between the two groups with regard to the duration of surgery (180 vs. 171 min), the number of reoperations for bleeding (4.7 % vs. 5.0 %), ventilation times, length of stay in the intensive care unit (2.6 vs. 2.4 days) or 30-day mortality (2.0 % vs. 2.8 %). The incidence of diabetes and occurrence of sternal dehiscence was higher in group A (1.6 % vs. 0.7 %; P < 0.01). Sternal complications were strongly associated with diabetes and the combined use of bilateral ITA grafts but not with isolated obesity. The incidence of sternal dehiscence was associated with the male gender (group A 1.9 % vs. 0.7 %; group B 0.9 % vs. 0.3 %). CONCLUSION: Severe obesity does not PER SE enhance perioperative mortality. A BMI of 30 to 50 combined with diabetes mellitus and bilateral ITA grafting increases the risk for sternal complications.

Patency of internal thoracic artery compared to vein grafts - postoperative angiographic findings in 1189 symptomatic patients in 12 years.

BACKGROUND: The evidence respecting the superior patency of ITA-grafts compared to SVG is conclusive. This study evaluates the angiographic findings in 1,189 symptomatic patients who received either one or both ITA-grafts with or without additional SVG. METHODS: 1,189 patients (975 males, aged 62.4 +/- 9.1 years), operated between 2/93 and 7/05, underwent angiography due to reappearance of angina. Data were compared for patency of single ITA (n = 618), bilateral ITA (n = 416) or SVG (n = 2,218). Re-catherization was performed after 3.8 +/- 2.7 years. Severe bypass stenosis or graft occlusion was related to the target vessels. RESULTS: 3,668 bypasses were performed in 1189 patients. The occlusion rate was 16.5 % for SVG and 7.0 % for all ITAs ( P < 0.001). Severe stenosis was detected in 4.8 % of SVG and in 3.7 % of ITAs ( P < 0.05). Patency was 89.6 % for LITA, 88.7 % for RITA, and 78.7 % for SVG. The occlusion rate for LITA was: to LAD 6.6 %, DIA 8.5 %, obtuse marginal branch/CX 11.5 %. The occlusion rate for RITA was: to LAD 4.6 %, RCA 9.1 %., diag. branch 7.1 %. The occlusion rate for vein grafts was: to LAD 17.3 %, DIA 14.4 %, obtuse marginal branch/CX 15.9 %, to RCA 17.0 %. Patency for all ITAs was 89.3 % vs.78.7 % for all SVG ( P < 0.05). Despite symptoms, bypass patency was found in 711 patients (59.8 %). CONCLUSION: The superior patency of ITA-grafts could be documented angiographically in a negatively selected, symptomatic population. Graft occlusion was at least twofold higher for SVG.

Aflatoxin M1 absorption and cytotoxicity on human intestinal in vitro model.

Aflatoxin M1 (AFM1) is the principal hydroxylated Aflatoxin B1 (AFB1) metabolite and is detected in milk of mammals, after consumption of feed contaminated with AFB1. As it is classified as probable human carcinogen (group 2B of the IARC), most countries have regulated its maximum allowed levels in milk in order to reduce AFM1 risk (50 ng/kg the EU and 500 ng/kg in the USA). It was demonstrated that if AFB1 must be converted into its reactive epoxide to exert its effects, and the protein binding may play an important role in its cytotoxicity. Conversely, the AFM1 epoxidation in human liver microsomes is very limited and studies with human cell line (MCL5), expressing or not expressing cytochrome P450 enzymes, demonstrated a direct toxic potential of AFM1 in absence of metabolic activation. For this reason, while AFM1 is generally considered a detoxification product of AFB1 relatively to carcinogenicity and mutagenicity property, this is not always true for cytotoxicity activity. Aim of this work is to evaluate the intestinal absorption of AFM1 using a human in vitro model, the Caco-2 cell line. Either the parental Caco-2 cell line or its derived clone TC7, with higher metabolic competence, have been used. They were treated with different concentrations of AFM1, that mirror the milk contamination level (0.3-32 nM corresponding to 10-10,000 ng/kg), either in undifferentiated or in differentiated phase of growth. After 48 h of treatment in serum free medium, a dose dependent absorption of AFM1 has been detected in both cell lines, especially in differentiated cells, while, no appreciable effects on cell viability were observed, except for a general cellular suffering, revealed by LDH release, particularly evident in the undifferentiated cells. As well, no metabolites or AFM1 conjugates have been detected. The present results may be crucial for the evaluation of human risk to AFM1 exposure, in particular for children's population, due to their large use of milk and derivatives.

The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics.

The human intestinal Caco-2 cell line has been extensively used over the last twenty years as a model of the intestinal barrier. The parental cell line, originally obtained from a human colon adenocarcinoma, undergoes in culture a process of spontaneous differentiation that leads to the formation of a monolayer of cells, expressing several morphological and functional characteristics of the mature enterocyte. Culture-related conditions were shown to influence the expression of these characteristics, in part due to the intrinsic heterogeneity of the parental cell line, leading to selection of sub-populations of cells becoming prominent in the culture. In addition, several clonal cell lines have been isolated from the parental line, exhibiting in general a more homogeneous expression of differentiation traits, while not always expressing all characteristics of the parental line. Culture-related conditions, as well as the different Caco-2 cell lines utilized in different laboratories, often make it extremely difficult to compare results in the literature. This review is aimed at summarizing recent, or previously unreviewed, data from the literature on the effects of culture-related factors and the influence of line sub-types (parental vs. different clonal lines) on the expression of differentiation traits important for the use of Caco-2 cells as a model of the absorptive and defensive properties of the intestinal mucosa. Since the use of Caco-2 cells has grown exponentially in recent years, it is particularly important to highlight these methodological aspects in order to promote the standardization and optimisation of this intestinal model.

Absorption of fumonisin B1 and aminopentol on an in vitro model of intestinal epithelium; the role of P-glycoprotein.

The aim of the present paper is to evaluate the absorption of fumonisin B1 and its principal metabolite, aminopentol on a human intestinal model, Caco-2 cells, cultured on semi-permeable inserts, that reproduces the two different intestinal compartments: luminal (apical) and serosal (basolateral) side. Following separate exposure in apical and in basolateral compartments, aminopentol passage through the cell layer (in particular from basolateral to apical direction) was shown, while it was not observed for the parent compound. The different aminopentol distribution between the two compartments of the culture system, and its variation in presence of verapamil or probenecid (P-gp and MRP inhibitors respectively), strongly suggests the involvement of P-glycoprotein in the influx/efflux mechanisms of aminopentol in the intestinal cells, reducing its oral bioavailability.

Double thoracic artery--halved mid-term mortality? A 5-year follow-up of 716 patients receiving bilateral ITA versus 662 patients with single ITA.

UNLABELLED: The superior patency of ITA grafts to saphenous veins is conclusive. The aim of the study was to collate mid-term benefit between patients receiving bilateral ITA (BITA) or single ITA (SITA). Outcome of 1378 pts with isolated CABG operated between 1/97-8/99 was analyzed retrospectively. Follow-up was 4.0 to 6.6 years (average 5.3). A total of 716 pts received BITA, 662 SITA and additional saphenous veins. We evaluated mortality rate, freedom from reoperation, intervention (PTCA/stent), and incidence of cardiac events and quality of life with respect to pts risk factors. Demographic data: Male gender was more frequent in both groups (BITA females: n=115; males: n=601; SITA females: n=150; males: n=512; p<0.01). Mean age was comparable in both groups with 69.2 years (42.7 to 88.6 years) in the BITA group and 71.0 years (47.3 to 91.6 years) (n. s.) in the SITA group. RISK FACTORS: Incidence of diabetes mellitus (26.0 vs 25.9%) as well as the mean BMI (27.4 vs 27.0%) did not differ statistically in both groups. RESULTS: Clinical characteristics like NYHA/ CCS classifications showed a significant difference towards superior results only for stadium I in the BITA group. Mortality/cardiac events after 5.3 years average: Total mortality revealed 5.2% (n=37) in the BITA vs 9.1% (n=60) in the SITA group (p

Is there a difference between diabetic and non-diabetic ITAs? Histomorphological and immunohistochemical examinations of internal thoracic arteries.

BACKGROUND: The relative resistance to arteriosclerosis of the internal thoracic artery is clinically valid. The aim of this study was to evaluate the graduation of intima and media fibrosis in ITA parts and to determine the extent of atherosclerosis using immunohistochemical methods related to patients' risk factors. METHODS: 227 distal ITA segments from 144 male and 83 female patients were examined. Mean age was 66.7 +/- 9.03 years. HE and Elastica van Gieson staining was performed to detect intima-/mediafibrosis. POPULATION: insulin-dependent diabetes mellitus (idDM; n = 35), non-insulin-dependent diabetes mellitus (nidDM; n = 32), irradiation of the chest due to cancer (n = 27), control group with isolated hyperlipidemia, hypertension, peripheral vascular disease (n = 133). 12 ITA segments of each group were examined immunohistochemically using CD34, CD68, Tenascin, Collagen III, Collagen IV. RESULTS: Histomorphological examinations using HE and Elastica van Gieson stainings showed for idDM-ITAs no fibrosis in 57 %, slight fibrosis in 34 %, medium in 6 %, no severe, for nidDM-ITAs no fibrosis in 63 %, slight 31 %, medium 3 %, severe 3 %, for ITAs after irradiation: no fibrosis in 37 %, slight in 62 %, no medium or severe fibrosis. Compared to the control group there was no statistically significant difference in diabetic ITAs. Immunohistochemical examinations, scored on a 0 - 3 basis with 0 being the lowest (no reaction), 3 the highest (severe reaction), showed mostly no or slight reaction to CD34, CD68, Tenascin, and Collagen III for intima and media, with comparable results for diabetic or "irradiated" ITAs to those of the control group. The most distinct reaction (medium= 2), was detected for Collagen IV, a marker which outlines basement membranes of endothelia and smooth muscle cells of the vessel wall, but without differences between the four groups. No severe reactions were observed. CONCLUSIONS: The relative resistance of ITA to arteriosclerosis could be demonstrated in 227 ITA segments. Immunohistochemistry supports histomorphological findings. The influence of diabetes mellitus and irradiation remains irrelevant.

Are men treated better than women? Outcome of male versus female patients after CABG using bilateral internal thoracic arteries.

BACKGROUND: Apart from smaller native coronary arteries a lower number of anastomoses and a lower ITA incidence have been suggested as being responsible for the increased mortality in female patients compared to males. The aim of this study was to clarify whether the outcome of females might be a consequence of a different regimen in anastomosis and ITA frequency. METHODS: We evaluated operative data and early outcome of 6906 patients with isolated CABG operated between 1/96 - 7/03 3064 out of 5381 males received bilateral ITA (BITA) vs. 750 of 1525 females. Single ITA (SITA) was performed in 2126 males and 704 females. RESULTS: Demographic and operative data: average age for male patients was 64.0 +/- 9.2 years, for females 68.5 +/- 8.6 years ( p < 0.05). The prevalence of diabetes mellitus was significantly higher in females (34.6 % vs. 27.4 %, p < 0.01). Body mass index (BMI, 26.6 vs. 27.4 mean), incidence of main stem stenosis (23.0 % vs. 23.5 %), ejection fraction < 40 % (7.8 % vs. 10.1 %), urgent or emergent operations (13.1 % vs. 11.3 %) and number of performed anastomoses (3.2 vs. 3.5 mean) showed no significant difference between males and females. Total ITA frequency did not differ (95.3 % vs. 96.5 %), but BITA frequency was significantly higher (56.9 % vs. 49.2 %, p < 0.01) in male patients. Overall 30-day mortality was 2.8 % for males vs. 4.1 % ( p < 0.05) for females. Cardiac-related mortality was significantly higher in female patients (2.6 % vs. 1.1 %, p < 0.01). Non-cardiac-related mortality did not differ significantly. Graft-related mortality for males and females revealed 2.7 % in the BITA, 3.3 % in the SITA group and 6.9 % for patients without ITAs and reached statistical significance ( p < 0.01) for SITA or BITA vs. the no-ITA group, but not for BITA vs. SITA grafting. Nevertheless cardiac-related mortality in male and female patients without an ITA graft was more than two-fold higher compared to these with single ITAs and more than three-fold higher compared to those with BITA grafting. CONCLUSIONS: Female gender, frequently associated with diabetes mellitus, presents a predictor for increased mortality in CABG. A discrimination of women with respect to a restriction of ITA grafting could be confirmed only for bilateral ITAs. The superior results of bilateral ITA grafts are independent of gender.

[Urgent or emergent coronary revascularization using bilateral internal thoracic artery after previous clopidogrel antiplatelet therapy]. / Dringliche oder notfallmässige Koronarrevaskularisation unter Verwendung beider Arteria Thoracica interna nach vorheriger Thrombozytenaggregationshemmung mit Clopidogrel.

BACKGROUND: Application of clopidogrel before diagnostic or therapeutical percutaneous coronary interventions has become standard for stent-thrombosis prevention. The irreversible platelet inhibition causes increasing bleeding complications if urgent coronary artery bypass grafting becomes necessary. This study evaluates the effect on bleeding complications of clopidogrel in urgent CABG using bilateral internal thoracic artery (ITA) and saphenous veins in all patients. METHODS: We retrospectively analyzed 166 patients (operated between 1/00-12/02) with urgent or emergency CABG, using both ITAs and compared 83 patients with previous (within 5 days) clopidogrel and aspirin application to 83 patients without clopidogrel. We evaluated chest tube output, reexploration rate and necessity of blood products, ventilation time and ICU stay. RESULTS: Both groups were comparable with age, gender, number of performed anastomoses (mean 4/ patient). Chest tube output (24 h) was higher in the clopidogrel group (935 +/- 599 ml vs 754 +/- 335 ml (p = 0.018)), as well as reexploration rate with 7.2% (6 of 83) vs 0% (0 of 83) (p < 0.001). Number of blood products in the clopidogrel group for red cells was 2.41 +/- 1.88 U vs 1.84 +/- 1.47 U p = 0.03, for plateletes 0.43 +/- 0.88 U vs 0.024 +/- 0.22 p = 0.0001, for fresh frozen plasma 0.41 +/- 1.14 U vs 0.096 +/- 0.59 U p = 0.029. Mechanical ventilation time was 11.35 +/- 8.77 h vs 10.57 +/- 9.12 h p = 0.51, ICU stay 32.1 +/- 21.8 h vs. 29.8 +/- 21.1 h (p = 0.48). CONCLUSIONS: Previous application of clopidogrel in combination with aspirin before urgent CABG induces increased chest tube output, reexploration rate and necessity of blood products, especially of plateletes. Nevertheless, routine use of both ITAs in patients after clopidogrel exposure can be performed with acceptable bleeding complications.

Toxicology investigations with cell culture systems: 20 years after.

From almost 20 years the "in vitro" model has gained a wide ground in toxicological investigation, providing advanced tools, reliable protocols, mechanistic information. These advancements have been done thanks to different approaches, addressed at improving chemical testing and validating procedures, at exploring the cellular and molecular basis of toxicity, at studying the modifications that xenobiotics undergo in the cellular environment. In this review the most advanced cellular models, the mechanisms of cell death, the techniques to monitor gene activation, following chemical exposure, is highlighted. Moreover the more recent in vitro models to approach the biotransformation issue will be presented.

[Simultaneous carotid endarterectomy and cardiac surgery-additional risk factor or safe procedure?]. / Thrombendarteriektomie der Arteria carotis und simultane Herzoperation. Zusätzlicher Risikofaktor oder sichere Vorgehensweise?

BACKGROUND: The occurrence of severe carotid artery disease in more than 12% of patients requiring ACB results in a discrepancy concerning the best treatment for both diseases. We reviewed the early outcome of patients with ACB and/or valve replacement and simultaneous carotid endarterectomy (TEA). METHODS: We evaluated retrospectively 244 patients operated simultaneously between 7/94 and 10/2001: 209 patients received ACB, 35 patients ACB and/or valve replacement. Mean age was 68 years. 188 patients were male. We analyzed risk factors, morbidity, incidence of neurological complications and 30 day mortality. RESULTS: Perioperative stroke with hemiplegia occurred in 3.3% (8 patients). Of these patients, 4 showed contralateral carotid artery occlusion, 2 contralateral severe stenosis. Two patients (0.8%) experienced prolonged reversible ischemic neurological deficit (PRIND), 4 patients (1.6%) transient ischemic attack (TIA). 30-day morbidity was 4.5%. Three patients died due to low cardiac output, 6 patients due to extracardial reasons, 2 patients (0.8) due to cerebral death. CONCLUSION: Simultaneous TEA and cardiac surgery can be performed with an acceptable risk for neurological complications and mortality. Occlusion of the contralateral carotid artery could be identified as an evident predictor for increased neurological complications. Compared to two-stage procedures, combined operations yield a reduction of hospital costs.

[End of the millennium - end of the single mammary artery graft? Two internal mammary arteries - standard for the next millennium? Early clinical results and analysis of risk factors in 1487 patients with bilateral internal mammary artery bypass]. / Ende eines Jahrtausends--Ende des Einfach-Mammaria-Bypasses. Zwei Arteriae mammariae internae--Standard für das nächste Jahrtausend? Klinische Ergebnisse und Risikofaktorenanalyse von 1487 Patienten mit bilateralem Arteria mammaria interna Bypass.

OBJECTIVE: CABG with bilateral IMA grafts (BIMA) can improve long-term results in cardiac morbidity and mortality. An enhanced incidence of bleeding and wound complications compared to patients with single IMA (SIMA) remains a matter of debate. The aim of the study was to compare the operative outcomes of patients who had undergone CABG with BIMA and SIMA in situ grafts, especially to identify patient-related risk factors, such as obesity, diabetes mellitus and age above 70 years. METHODS: Out of a total of 5144 patients operated on between January 1996 and September 1999, patients with isolated CABG (n = 3671) with BIMA or SIMA were analyzed retrospectively. In the BIMA group, the patients' (n = 1478) mean age was 64.0 years; mean EF was 62.1%. In the SIMA group (n = 2184), mean age was 65.4 years and mean EF was 60.6% (n.s.). In the BIMA group, the right IMA was led anterior of the aorta to the LAD, the left IMA to the lateral wall. In the SIMA group, the LAD was revascularized with the left IMA. Additional bypasses were performed with vein grafts. RESULTS: The 30-day mortality was 1.6% in the BIMA group, 1.7% in the SIMA group in patients under 70 years, and 4.1% (BIMA) and 4.0% (SIMA) in patients over 70 years (p = n.s.). A significantly higher blood loss was observed in the BIMA group (BIMA 979 +/- 708 ml; SIMA 790 +/- 575 ml; p < 0.05). The rethoracotomy rate due to bleeding was significantly higher in patients with BIMA (4.1%) compared to those with SIMA (2.5%; p < 0.05). In patients with a body mass index (BMI) of less than 27, no significant difference could be found (SIMA 2.8%, BIMA 3.4%; p = n.s.). Patients with a BMI > 27 showed a significantly higher rethoractomy rate (SIMA 2.2%; BIMA 4.9%). A higher incidence of sternal instabilities could be observed in the BIMA group (4.2%; p < 0.05). Diabetes mellitus could not be identified as an independent risk factor for sternal complications (SIMA 2.9%; BIMA 5.0%; p = n.s.). CONCLUSION: CABG using both IMAs can be performed in nearly all patients as a routine method with good clinical results and low mortality. Bleeding in the BIMA group within 48 h was increased. BMI > 27 could be identified as a risk factor for sternal complications, but not diabetes mellitus or age over 70 years.

Urgent or emergent coronary revascularization using bilateral internal thoracic artery after previous clopidogrel antiplatelet therapy.

BACKGROUND: Clopidogrel application before diagnostic or therapeutical percutaneous coronary intervention has become the standard for stent thrombosis prevention. Irreversible platelet inhibition causes increasing bleeding complications if urgent coronary artery bypass grafting is necessary. This study evaluates the effect on bleeding complications of clopidogrel in urgent CABG using bilateral internal thoracic artery (ITA) and saphenous veins in all patients. METHODS: We retrospectively analyzed 128 patients (operated between January 2000 and September 2002) undergoing urgent or emergent CABG using both ITAs, and compared 64 patients with previous clopidogrel and aspirin application (within 5 days) to 64 patients without clopidogrel. We evaluated chest tube output, re-exploration rate and necessity of blood products, ventilation time and ICU stay. RESULTS: Both groups were comparable in age, gender, number of performed anastomoses (mean 4/patient). Chest tube output (24 h) was higher in the clopidogrel group at 977+/-628 ml vs. 788+/-389 ml (p=0.046), as was re-exploration rate with 7.81% (5 of 64) vs. 0% (0 of 64) (p<0.005). The number of blood products amounted to 2.7+/-1.9 U in the clopidogrel group vs. 1.9+/-1.6 U (p=0.013) for red cells, 0.05+/-0.9 U vs. 0.03+/-0.25 (p=0.0003) for platelets, and 0.5+/-1.3 U vs. 0.2+/-1.0 U (p=0.14) for fresh frozen plasma. Mechanical ventilation time was 11.9+/-9.7 h vs. 9.6+/-5.9 h (p=0.10), ICU stay 32.6+/-22.1 h vs. 27.8+/-18.2 h (p=0.19). CONCLUSIONS: Previous application of clopidogrel in combination with aspirin before urgent CABG induces increased chest tube output, re-exploration rate and necessity of blood products, especially platelets. Nevertheless, routine use of both ITAs in patients after clopidogrel exposure can be performed with acceptable bleeding complications.

Simultaneous carotid endarterectomy and cardiac surgery--additional risk factor or safety procedure?

BACKGROUND: The occurrence of severe carotid artery disease in more than 12 % of patients requiring CABG results in a discrepancy concerning the best treatment for both diseases. We reviewed the early outcome of patients with CABG and/or valve replacement and simultaneous carotid endarterectomy (CEA). METHODS: We evaluated retrospectively 244 patients operated simultaneously between 7/94 and 10/2001. 209 patients received CABG; 35 patients CABG and/or valve replacement. Mean age was 68 years. 188 patients were male. We analyzed risk factors, morbidity, incidence of neurological complications and 30 day mortality. RESULTS: Perioperative stroke with hemiplegia occurred in 3.3 % (8 patients). 4 of these patients showed contralateral carotid artery occlusion, 2 contralateral severe stenosis. 2 patients (0.8 %) experienced prolonged reversible ischemic neurological deficit (PRIND), 4 patients (1.6 %) transient ischemic attack (TIA). 30-day lethality was 4.5 %. 3 patients died due to low cardiac output, 6 patients due to extracardial reasons, 2 patients (0.8 %) developed a cerebral death. CONCLUSIONS: Simultaneous CEA and cardiac surgery can be performed with an acceptable risk for neurological complications and mortality. Occlusion of contralateral carotid artery could be identified as an evident predictor for increased neurological complications. Compared to two-stage procedures, combined operations yield a reduction of hospital costs.

Evaluation of Fumonisin B(1) and its metabolites absorption and toxicity on intestinal cells line Caco-2.

The aim of the present paper is to investigate intestinal absorption and toxicity of Fumonisin B(1) (FB(1)) and its partially (PHFB(1) and PHFB(2)) and totally hydrolyzed (HFB(1)) metabolites, using the human intestinal cell line Caco-2, a very well known in vitro model of intestinal epithelium for absorption and metabolism studies. Caco-2 cells were treated for 48 h with several toxin concentrations (in the range of 1-138 microM). At the end of exposure period, no significant variation on cell viability has been observed with all chemicals tested, either in undifferentiated cells or in differentiated ones, suggesting a poor toxicity of these mycotoxins for intestinal cells. In any case, FB(1) appears the most active in this respect. For which concerns the cellular absorption, FB(1), PHFB(1) and PHFB(2) are never detected into Caco-2 cells. On the contrary, a dose-dependent absorption of HFB(1) has been observed in differentiated cells, which express enzymatic and metabolic characteristics of mature enterocytes. Thus HFB(1), losing the tricarballylic acid chain, is more bioavailable than FB(1) on intestinal cell, supporting the hypothesis that in risk evaluation of fumonisins exposure its metabolites are also relevant.

Identification and coupling to adenylate cyclase of three different [(3)H]CGP 12177 binding sites in Caco-2 cell membranes.

In the present investigation the identification of beta -adrenoceptor (beta -ARs) subtypes in the Caco-2 cell line was performed using radiometric assays. beta -ARs were measured using increasing concentrations of the highly specific beta -AR antagonist (-)[(3)H]CGP 12177 (0.06-4 nM), whereas the beta(1)- and beta(2)-AR subtypes discriminated through selective binding assays using the highly selective unlabelled antagonists CGP 20712A and ICI 118551. Atypical beta -ARs were measured using an incubation system formed by higher concentrations (0.6-20 nM) of (-)[(3)H]CGP 12177. beta - Atypical binding site concentrations (69 +/- 5 fmol mg ml(-1)of membrane protein) were higher than beta(1)-ARs (7 +/- 1) and beta(2)-ARs (24 +/- 2), respectively. The different beta -AR subtype affinities were characterized by binding inhibition experiments and the adrenergic agonists displaced the radioligand from its specific binding sites in the following order of potency: isoproterenol > clenbuterol > dobutamine > SR 58611A; for antagonists the order of potency was: propranolol approximately = ICI118551 approximately = CGP20712A. For atypical beta -ARs the order was: SR 58611A > clenbuterol > dobutamine > isoproterenol for agonists and propranolol > CGP 20712A > ICI 118551 for antagonists. As far as in vitro functional studies are concerned, beta -AR subtypes were shown to be coupled to adenylyl cyclase as their stimulation produced cAMP in an amount significantly higher than basal values. cAMP production after stimulation with dobutamine, clenbuterol, isoproterenol, and SR 58611A was measured using a cAMP radioassay kit. The order of efficacy suggested that the stimulation of beta(2)-ARs was the most effective in inducing the activation of cell signalling mechanisms. The identification of functional beta -ARs in a cancer cell line represents the first step in the study of the possible adrenergic control of cellular activities (e.g. proliferation and/or differentiation), which could suggest the use of this cancer cell line as a model for the study of cell activity or possibly new therapeutic strategies.