Physical and Chemical Effects of Different Working Gases in Coffee Brewing: A Case Study of Caffè Firenze.

(1) Background: Recently, a new espresso extraction method, Caffè Firenze, has been developed, which uses gas at operating pressures of 20 bar to obtain abundant, persistent foam. The experiment aimed to evaluate the effect of using six gases (air, argon, nitrogen, carbon dioxide, carbon/nitrogen mix, and nitrous oxide) on the foam and liquid coffee. (2) Methods: Foam volume, persistence, sugar retention time, color, and rheological properties were measured. Volatile organic compounds were also evaluated. Analyses were also carried out on the liquid coffee to determine caffeine and chlorogenic acid concentrations. (3) Results: The analysis of variance revealed significant differences between the gases for all parameters. Multivariate analysis identified three groups of gases: the first comprised air, N2, and Ar; the second CO2 and N2O; and the third comprised samples extracted with CO2/N2 mix. (4) Conclusions: The choice of gas significantly influences the drink's chemical-physical characteristics and is fundamental for product diversification.

Effects of a Small Increase in Carbon Dioxide Pressure during Fermentation on Wine Aroma.

The present study tested the effect of a slight increase in pressure (from 0 to 1 bar) during the fermentation on the wine aroma profile. Fermentations were carried out with a commercial dry yeast on Sangiovese juice in the absence of berry skins. The wine samples fermented under slight overpressure conditions were found to be significantly different from the control samples produced at atmospheric pressure in relation to several chemical compounds. Concentrations of many esters (i.e., isoamyl acetate, ethyl acetate, ethyl hexanoate, hexyl acetate, ethyl dodecanoate, and ethyl tetradecanoate), and acids (i.e., hexanoic acid, octanoic acid, and decanoic acid) increased, while concentrations of two acids (i.e., isobutyric and isovaleric acid) decreased. These differences, notably the higher concentration of esters, are usually associated with a more intense fruity attribute. Triangular sensory tests revealed that the significant chemical differences were also perceivable; hence, introducing a slight pressure increase during the alcoholic fermentation could be a useful tool in managing the aroma profile of wine.

Filtration Scheduling: Quality Changes in Freshly Produced Virgin Olive Oil.

Filtration is the most widespread stabilisation operation for extra virgin olive oil, preventing microbial and enzymatic changes. However, during the harvest, the workload of olive mills is at its peak. This results in two approaches to filtration: (i) delays it until after harvesting, increasing the risk of degraded oil quality, and (ii) filters it immediately, increasing the workload. The aim of our experiment is to assess the risk of delaying filtration and establish a safe delay time. Changes in the sensory profile and volatile compound contents were evaluated during 30 days in filtered and unfiltered samples. Significant differences were related to filtration: both turbidity grade and microbial contamination; no differences for the legal parameters were found. Two, contrasting, results were obtained with respect to oil quality: (i) the fusty defect, appearing in less than five days in unfiltered oils, leading to the downgrade of the oil's commercial category, and (ii) filtration removing some lipoxygenase volatile compounds. Consequently, a fruity attribute was more pronounced in unfiltered samples until day five of storage; it seems that, from this point, the fusty defect masked a fruity attribute. Hence, filtering within a few days strongly reduced the risk of degraded oil quality compared to a delayed filtration.

Short-term prognosis of unstable angina in the era of high-sensitivity cardiac troponin: insights for early rule-out strategies.

BACKGROUND: It is unclear if strategies to rule-out myocardial infarction (MI) based on a single high-sensitivity troponin T (hsTnT) measurement at the emergency department (ED) presentation may also exclude unstable angina. METHODS: We measured hsTnT ex-post on the admission frozen blood sample of 644 subjects with Braunwald IIIB CK-MB-negative unstable angina. This analysis included the 240 patients with hsTnT value ≤99th percentile reference limit (UA). We evaluated the clinical outcome of UA patients and the applicability of two rule-out strategies based on the combination of a non-ischemic ECG with (1) a single hsTnT value below the Limit-of-Detection (LoD), (2) a TIMI risk score ≤1. RESULTS: UA patients with hsTnT ≤99th percentile reference limit had a favorable 30-day outcome [0.8% MI, 0% cardiovascular death (CVD)], but the rate of CVD/MI at 180-day was 4.7%. Sensitivities for UA were 94.6% according to the 'TIMI ≤1-strategy' and 75.4% according to 'hsTnT-below-LoD-strategy', accounting for 5.4 and 24.6% missed diagnoses, respectively. A prognostic risk stratification to guide appropriate outpatient assessment in potential discharged unrecognized UA patients was developed: a risk score based on the combination of age >60 years and C-reactive protein >4.5 mg/L effectively stratified the 180-day CVD/MI occurrence: 0, 2.5 and 12.7% for score 0, 1 and 2 (log-rank = 0.001, C-statistic = 0.776). CONCLUSION: Single measurement hsTnT strategies, successfully tested to rule-out MI, may allow safe ED discharge of patients with a suspected acute coronary syndrome: even if UA may not be excluded, its short-term prognosis is favorable. UA patients with a C-reactive protein >4.5 mg/L and older than 60 years have a substantial medium-term cardiovascular risk and may benefit from a timely outpatient diagnostic assessment.

Understanding Olive Oil Stability Using Filtration and High Hydrostatic Pressure.

Veiled extra virgin olive oil (VEVOO) is very attractive on the global market. A study was performed to highlight the role of different amounts of water and microorganisms on the evolution of VEVOO quality during storage, using the selective effects of the application of individual or combined filtration and high hydrostatic pressure (HHP) treatments. Four oil processing trials were carried out in four replicates, resulting in a full factorial design with two independent fixed factors: filtration and HPP treatments. The turbidity of all the olive oil samples was characterized. Furthermore, all the olive oil samples were analysed for legal parameters, volatile organic compounds and phenolic compounds during the storage tests. The microbial contamination in the presence of a high level of water activity (>0.6 Aw) was related to the formation of volatile aroma compounds, which were responsible for the "fusty" sensory defect. Furthermore, high water activity values were related to an increase in the hydrolytic degradation rate of the phenolic compounds. The oil turbidity has to be planned and controlled, starting from adjustment of the water content and application of good manufacturing practices.

What kind of coffee do you drink? An investigation on effects of eight different extraction methods.

The chemical composition of brewed coffee depends on numerous factors: the beans, post-harvest processing and, finally, the extraction method. In recent decades, numerous coffee-based beverages, obtained using different extraction techniques have entered the market. This study characterizes and compares eight extraction coffee methods from a chemical-physical point of view, starting from the same raw material. Specifically, three types of Espresso, Moka, French Press, and 3 filter coffee that for the first time are reported in the scientific literature Cold Brew, V60, and Aeropress are compared. Physical measurements included the quantification of total dissolved solids, density, pH, conductivity, and viscosity. Chemical analyses identified 15 chlorogenic acids (CGAs): six caffeoylquinic acids, one p-Coumaroylquinic acid, one Feruloylquinic Acid, four Caffeoylquinic lactones, and three Dicaffeoylquinic acids. Maximum caffeine and CGA concentrations were found in Espresso coffees, while Moka and filtered coffees were three to six times less concentrated. The classic Espresso method was most efficient for caffeine and CGA recovery, with a yield almost double that of other methods. Per-cup caffeine and CGAs were higher in Cold Brew than Espresso coffees, as a function of the volume of beverage, which ranged from 30 mL (for espresso) to 120 mL (for filtered coffees). In light of these results, it is not possible to establish how many cups of coffee can be consumed per day without exceeding the recommended doses, since according to the applied brewing method, the content of the bioactive substances varies considerably.

Characterization and comparison of cold brew and cold drip coffee extraction methods.

BACKGROUND: Each region of the world has its own methods, protocols, instruments and procedures regarding how to brew coffee. The final result in the cup is strongly affected by the extraction method, and many studies have focused on this subject. However, few studies have investigated slow, cold extraction methods, despite their popularity among baristas. Therefore, the present study aimed to characterize and compare two cold extraction methods: cold brew and cold drip. RESULTS: Physical and chemical analyses were used to describe coffee beverages in terms of pH, total solids, refractive index, density and viscosity. Caffeine and cinnamic acids were quantified using high-performance liquid chromatography (HPLC)/diode array detector and HPLC/mass spectrometry. A sensory evaluation included aroma, flavor and textural attributes. CONCLUSIONS: Significant differences were found in the chemical and physical parameters, both between and within the two methods, as a function of the extraction temperature and contact time. Similarly, the sensory evaluation found differences in flavor profiles, as measured in terms of bitterness, sweetness, sourness and global intensity. © 2018 Society of Chemical Industry.

Prognostic implications of high-sensitivity cardiac troponin T assay in a real-world population with non-ST-elevation acute coronary syndrome.

BACKGROUND: High-sensitivity cardiac troponin T (hsTnT) was recently approved for clinical use by the Food and Drug Administration. The transition from contemporary to hsTnT assays requires a thorough understanding of the clinical differences between these assays. HYPOTHESIS: HsTnT may provide a more accurate prognostic stratification than contemporary cardiac troponin I (cTnI) in patients with non-ST-segment elevation acute coronary syndromes (NSTE-ACS). METHODS: HsTnT and cTnI were measured in 644 patients with CK-MB negative NSTE-ACS who were enrolled in the prospective multicenter SPAI (Stratificazione Prognostica dell'Angina Instabile) study. Patients were stratified at the 99th percentile reference limit for each assay. The primary endpoint was cardiovascular death (CVD) or non-fatal myocardial infarction (MI); the secondary endpoint was the occurrence of unstable angina (UA). Follow-up lasted 180 days. RESULTS: Patients with hsTnT ≥99th percentile were at higher risk of CVD/MI (30-day: 5.9% vs 0.8%, p = 0.001; 180-day: 11.1% vs 4.7%, p = 0.004), also after adjusting for TIMI Risk Score. No significant difference in CVD/MI at 180-day was found between hsTnT-positive/cTnI-negative and hsTnT-negative/cTnI-negative patients (adjHR 1.61, 95% CI 0.74-3.49, p = 0.232). Occurrence of UA was not differently distributed between hsTnT groups dichotomized at the 99th percentile (12.4% vs 12.5% p = 0.54). CONCLUSIONS: Our investigation on a real-world NSTE-ACS population showed good prognostic performance of hsTnT in the risk stratification of the hard endpoint, but did not demonstrate the improved prognostic ability of hsTnT over contemporary cTn. Neither troponin assay predicted the recurrence of UA, suggesting the acute rise of cardiac troponin as a marker of severity, but not the occurrence of future coronary instability.

Serum uric acid on admission predicts in-hospital mortality in patients with acute coronary syndrome.

BACKGROUND: Despite the association between uric acid and cardiovascular disease has been known for decades, the prognostic value of serum uric acid (UA) in all clinical manifestations of acute coronary syndrome (ACS), namely ST-elevation myocardial infarction (STEMI), NSTEMI and unstable angina, has not been definitively assessed. METHODS: This retrospective analysis included patients from previous SPAI and FAMI studies with the aim to investigate the association between serum uric acid and major adverse cardiovascular events at 180days from hospital admission. RESULTS: 1548 patients were considered and divided in four groups, according UA concentration. Uricemia was significantly associated with gender, BMI, arterial hypertension, HDL-cholesterol, triglycerides, metabolic syndrome and glomerular filtration rate in univariate analysis. Multivariate logistic regression indicated that UA >6.0mg/dL on admission increased the risk of in-hospital mortality in overall population (OR 2.9, 95%CI 1.4-6.1; p=0.0057) and in patients with de novo ACS (OR 3.2, 95%CI 1.5-6.8; p=0.0033). Comparable results were also obtained after adjusting the model for age, gender, body mass index, glomerular filtration rate, metabolic syndrome, acute revascularization and ethnicity. A positive correlation was observed between UA and C reactive protein concentrations in in-hospital deaths only (rho 0.41, p=0.027). CONCLUSION: In patients with acute coronary syndrome, uricemia levels above the current international reference limit (6.0mg/dl) were associated with in-hospital mortality, independently from ethnicity and renal function.

Low molecular weight heparin (parnaparin) for cardioembolic events prevention in patients with atrial fibrillation undergoing elective electrical cardioversion: a prospective cohort study.

Systemic thromboembolism is a severe complication in patients undergoing electrical cardioversion (ECV) for atrial fibrillation (AF). Vitamin K antagonists greatly reduce the risk of thromboembolic events, but the administration scheme before ECV is troublesome as difficulties in reaching and maintaining the target therapeutic range for 3 weeks often delay the restoration and likelihood of maintaining sinus rhythm. Low molecular weight heparins (LMWHs) do not need dose adjustment, and may be preferable in this clinical setting. In this multicentre study, the LMWH parnaparin was used at a dose of 85 anti-factor Xa U/kg b.i.d. 2 weeks before and 3 weeks after ECV of AF. In an intention to treat analysis of 102 patients, there was no systemic thromboembolism or major bleeding (0%, 95% CI 0-3.6). Two clinically relevant non-major bleeds (2.5%, 95% CI 0.7-8.8) and three minor bleeds (3.8%, 95% CI 1.3-10.6) were recorded. No heparin-induced thrombocytopenia or other major adverse events were recorded. Parnaparin appears effective and safe for thromboprophylaxis of elective ECV in patients with AF.

Platelet-leukocyte mixed conjugates in patients with atrial fibrillation.

Although platelets may contribute to the inflammatory component in atrial fibrillation (AF), the impact of platelet-leukocyte mixed conjugates has not yet been determined. Seventeen patients with persistent AF (8/9 m/f; mean age 68.1 +/- 2.5 years), not on anticoagulant therapy, were recruited and compared to 34 healthy controls with normal sinus rhythm (16/18 m/f; mean age 60.8 +/- 1.2 years). Platelet-leukocyte mixed conjugates, platelet P-selectin and leukocyte activation markers (CD11b, myeloperoxidase) were measured by flow-cytometry in whole blood both in basal condition and after in vitro ADP/collagen challenge. Plasma D-dimer and soluble P-selectin were also measured. Statistical analyses were performed by Mann-Whitney or Wilcoxon U test for intergroup differences. In AF patients platelet count, as well as platelet P-selectin expression and percent platelet-leukocyte conjugates were all significantly lower both in basal condition and upon activation with ADP/collagen. In contrast, both soluble P-selectin and D-dimer were significantly higher than in controls; white blood cell count and leukocyte activation markers were unchanged. In conclusion, the formation of platelet-leukocyte mixed conjugates was unexpectedly reduced in AF, possibly due to less reactive platelets as a consequence of previous in vivo activation by ongoing formation of trace amounts of thrombin.

Prognostic value of heart rate turbulence and its relation to inflammation in patients with unstable angina pectoris.

Heart rate turbulence (HRT) provided insights into cardiac autonomic function and predicted clinical outcome in patients with myocardial infarction. A relation between cardiac autonomic function and inflammation was shown in several clinical settings. To assess the prognostic impact of HRT and its relation with inflammation in patients with unstable angina pectoris (UAP), HRT parameters (turbulence onset [TO] and turbulence slope [TS]) were measured in 331 patients with UAP (age 66.4 +/- 10 years; 231 men) with premature ventricular complexes on electrocardiographic Holter monitoring. Total and cardiac mortality were assessed at the 6-month follow-up. The 6th (-1.52%) and 4th deciles (4.90 ms/RR) were the best prognostic cut-off values for TO and TS, respectively. TS <4.9 ms/RR was associated with hazard ratio (HRs) of 7.10 (95% confidence interval [CI] 2.68 to 18.8, p = 0.0001) and 8.02 (95% CI 2.73 to 23.6, p = 0.0002) for total and cardiac mortality, respectively. The same HRs for TO >-1.52% were 2.94 (95% CI 1.11 to 7.81, p = 0.03) and 3.33 (95% CI 1.13 to 9.79, p = 0.029), respectively. Patients with TO <-1.52% and TS >4.9 ms/RR showed very low risks of total and cardiac mortality (1.8% and 0.9%, respectively). TS <4.9 ms/RR was independently associated with total (HR 3.87, 95% CI 1.21 to 12.3, p = 0.02) and cardiac (HR 3.81, 95% CI 1.01 to 14.4, p = 0.048) mortality at multivariable analyses. Both TS (r = -0.29, p <0.001) and TO (r = 0.16, p = 0.005) showed significant correlation with serum C-reactive protein. Thus, HRT can be helpful for risk stratification of patients with UAP. The association between cardiac autonomic function and inflammation can be pathogenetically relevant in this clinical setting.

Outcomes of the tacrolimus drug-eluting Janus stent: a prospective two-centre registry in high-risk patients.

BACKGROUND: To date, only two drug-eluting stents (DES) have been extensively tested in both randomized controlled clinical trials and large 'real world' registries: sirolimus-DES (Cypher stent, Cordis, Miami Lakes, Florida, USA) and paclitaxel-DES (Taxus stent, Boston Scientific, Natick, Massachusetts, USA). Recently, a new polymer-free tacrolimus-eluting Carbofilm-coated stent, the Janus stent (Sorin Biomedica, Saluggia, Italy), has entered the market but only few clinical data testing its safety and efficacy in selected patients are available. Thus, we performed a prospective registry of consecutive, unselected patients receiving this new DES. METHODS: A total of 118 patients scheduled for percutaneous coronary intervention (PCI) with DES were enrolled in two separate centers. End-points were: (i) immediate angiographic failure; (ii) major adverse coronary events (MACE) defined as the composite of cardiovascular death, nonfatal myocardial infarction (MI) and target lesion revascularization (TLR); (iii) clinically driven TLR; (iv) and stent thrombosis at 6-month follow-up. RESULTS: A total of 192 Janus stents were successfully implanted during elective (36%) or urgent PCIs (64%), including patients with ST elevation MI (16%). Twenty-four percent of patients were diabetics and 27% underwent multivessel PCI. Target lesions were B2-C type in 54%, in-stent restenosis in 8%, and located in degenerated venous grafts in 9%. Angiographic failure was observed in five of the 147 (3.4%) lesions treated. Total MACE rate at 6-month follow-up was 22% and clinically-driven TLR was carried out in 14% of patients. Stent thrombosis occurred in 4% of cases. CONCLUSION: This registry of the new tacrolimus-eluting Carbofilm-coated Janus stent showed an incidence of MACE, TLR and stent thrombosis higher than that reported in previous similar studies on DES. Whether this risk is due to this specific device or to the unselected (i.e. high-risk) population warrants further research.

Immunohistochemical analysis of pRb2/p130, VEGF, EZH2, p53, p16(INK4A), p27(KIP1), p21(WAF1), Ki-67 expression patterns in gastric cancer.

Although the considerable progress against gastric cancer, it remains a complex lethal disease defined by peculiar histological and molecular features. The purpose of the present study was to investigate pRb2/p130, VEGF, EZH2, p53, p16(INK4A), p27(KIP1), p21(WAF1), Ki-67 expressions, and analyze their possible correlations with clinicopathological factors. The expression patterns were examined by immunohistochemistry in 47 patients, 27 evaluated of intestinal-type, and 20 of diffuse-type, with a mean follow up of 56 months and by Western blot in AGS, N87, KATO-III, and YCC-2, -3, -16 gastric cell lines. Overall, stomach cancer showed EZH2 correlated with high levels of p53, Ki-67, and cytoplasmic pRb2/p130 (P < 0.05, and P < 0.01, respectively). Increased expression of EZH2 was found in the intestinal-type and correlated with the risk of distant metastasis (P < 0.05 and P < 0.01, respectively), demonstrating that this protein may have a prognostic value in this type of cancer. Interestingly, a strong inverse correlation was observed between p27(KIP1) expression levels and the risk of advanced disease and metastasis (P < 0.05), and a positive correlation between the expression levels of p21(WAF1) and low-grade (G1) gastric tumors (P < 0.05), confirming the traditionally accepted role for these tumor-suppressor genes in gastric cancer. Finally, a direct correlation was found between the expression levels of nuclear pRb2/p130 and low-grade (G1) gastric tumors that was statistically significant (P < 0.05). Altogether, these data may help shed some additional light on the pathogenetic mechanisms related to the two main gastric cancer histotypes and their invasive potentials.

Relation of heart rate variability to serum levels of C-reactive protein in patients with unstable angina pectoris.

Heart rate variability (HRV) and systemic markers of inflammation have prognostic value in patients with unstable angina pectoris (UAP). However, it is unknown whether any relation exists between HRV parameters and indexes of inflammation in this clinical context. We assessed HRV on 24-hour electrocardiographic Holter recordings, performed within 24 hours of admission, and measured C-reactive protein (CRP) serum levels by a high-sensitivity assay on admission, in 531 patients with UAP (65+/-10 years of age; 347 men) who were enrolled in the prospective multicenter study Stratificazione Prognostica dell'Angina Instabile (SPAI). A significant inverse correlation was found between CRP levels and all HRV parameters, with the highest r coefficient shown with SD of all RR intervals (r= -0.23; p<0.001) in the time domain and with very low-frequency amplitude (r= -0.22; p<0.001) in the frequency domain. When patients were categorized into 4 groups according to CRP quartile levels, statistically significant lower HRV values were observed in the upper CRP quartile. On separate multiple regression analyses, including the most important clinical and laboratory variables, SD of all RR intervals and very low-frequency amplitude were the most significant predictors of increasing CRP levels (p<0.001 for the 2 comparisons). In contrast, in models with SD of all RR intervals and very low-frequency amplitude as dependent variables, CRP was a strong predictor of impaired cardiac autonomic function (p<0.001 for the 2 comparisons). Thus, our data show that, in patients with UAP, high levels of serum CRP levels are significantly associated with decreased HRV, suggesting a possible pathophysiologic link between cardiac autonomic dysfunction and inflammatory activity.

Sirolimus-eluting vs uncoated stents for prevention of restenosis in small coronary arteries: a randomized trial.

CONTEXT: Percutaneous coronary revascularization of small vessels is associated with a high restenosis rate. Sirolimus-eluting stents reduce restenosis in simple and previously untreated lesions of large coronary arteries, but their outcomes in small vessels have not been adequately investigated. OBJECTIVE: To determine whether sirolimus-eluting stents are associated with a reduced 8-month rate of angiographic restenosis in comparison with an uncoated stent. DESIGN, SETTING, AND PATIENTS: This was a randomized, multicenter, single-blind, prospective trial performed with 257 patients undergoing percutaneous coronary revascularization for ischemic heart disease, and who had a previously untreated atherosclerotic lesion located in a small segment with a diameter of 2.75 mm or less, in 20 Italian centers between August 2002 and December 2003. INTERVENTION: Patients were randomly assigned to receive a sirolimus-eluting stent (129 patients) or an uncoated stent having an identical architecture and radiographic appearance (128 patients). MAIN OUTCOME MEASURES: The primary end point was the 8-month binary in-segment restenosis rate; secondary end points included procedural success and the 8-month rate of major adverse cardiac and cerebrovascular events. RESULTS: The mean (SD) reference diameter of the treated segment was 2.2 (0.28) mm; the lesion length, 11.84 (6.15) mm. After 8 months, the binary in-segment restenosis rate was 53.1% (60/113) in the patients receiving an uncoated stent and 9.8% (12/123) in those receiving a sirolimus-eluting stent (relative risk [RR], 0.18; 95% confidence interval [CI], 0.10-0.32; P<.001). Fewer patients randomized to sirolimus-eluting stents experienced major adverse cardiac events (12/129 [9.3%] vs 40/128 [31.3%]; RR, 0.30; 95% CI, 0.15-0.55; P<.001) mainly because of a reduction in target lesion revascularization (9/129 [7%] vs 27/128 [21.1%]; RR, 0.33; 95% CI, 0.14-0.70; P = .002) and myocardial infarction (2/129 [1.6%] vs 10/129 [7.8%]; RR, 0.20; 95% CI, 0.01-0.93; P = .04). CONCLUSION: The use of sirolimus-eluting stents to treat atherosclerotic lesions in small coronary arteries reduces restenosis and may also reduce major adverse cardiac events.

pRb2/p130, vascular endothelial growth factor, p27(KIP1), and proliferating cell nuclear antigen expression in hepatocellular carcinoma: their clinical significance.

Hepatocarcinoma (HCC) is the fifth most common cancer, with more than one million fatalities occurring annually worldwide. Multiple risk factors are associated with HCC disease etiology, the highest incidence being in patients with chronic hepatitis B virus and hepatitis C virus, although other factors such as genetic makeup and environmental exposure are involved. Multiple genetic alterations including the activation of oncogenes and inactivation of tumor suppressor genes are required for malignancy in human cancers and are correlated with increased stages of carcinogenesis and further tumor progression. In this study of 21 HCC patients, we analyzed pRb2/p130, vascular endothelial growth factor (VEGF), p27((KIP1)), and proliferating cell nuclear antigen as potential HCC molecular biomarkers. In our sample set, we found that p27((KIP1)) was absent. Univariate survival analysis showed that proliferating cell nuclear antigen expression (diffuse staining >50% of positive cells in tumor) was confirmed as a significant HCC prognostic biomarker for determining patient survival agreeing with previous studies (P = 0.0126, log-rank test). Lower pRb2/p130 expression was associated to a borderline P value of inverse correlation with tumor malignancy and to a positive correlation with respect to the time from HCC diagnosis (Spearman coefficient = 0.568; P < 0.05). Conversely, higher VEGF expression was associated with a poor survival (P = 0.0257, log-rank test). We demonstrate for the first time that pRb2/p130 is inversely correlated with VEGF expression and tumor aggressiveness (P < 0.05) in p27((KIP1))-negative HCC patients. pRb2/p130 and VEGF expression are independent from tumor staging, suggesting their possible role as independent prognostic molecular biomarkers in HCC. Furthermore, we have evidence that VEGF together with pRb2/p130 may act as new HCC biomarkers in a p27((KIP1))-independent manner. Additional studies with larger numbers of patient data would allow the use of multivariable techniques and would be able to further identify patients with poorer survival.

Circulating levels of cyclosporin A in inflammatory bowel disease: relationships with lymphocyte inhibition and the age of patients.

OBJECTIVE: To determine in the same blood sample the concentrations of cyclosporin A (CsA) and the degree of CsA-induced lymphocyte inhibition; to establish a relationship between these parameters; and to investigate the factor(s) influencing such a putative relationship. METHODS: Ten patients with a diagnosis of Crohn's disease (n=7) or ulcerative colitis (n=3) were enrolled in the study. The patients, who had never been immunosuppressed, were treated with microemulsion CsA twice daily by the oral route; at steady-state, blood samples were collected 0, 0.5, 1, 2, 3, 5, 7 and 12 h after the morning dose. CsA blood levels were measured by means of radioimmunoassay. The percentages of lymphocytes during the S-phase were assessed by flow-cytometry on the same blood specimens, only for samples collected at 0 h and 2 h. RESULTS: An inverse relationship emerged between CsA blood concentrations and the percentage of lymphocytes during the S-phase: the latter was maximal before the beginning of treatment and minimal in association with peak CsA levels. Furthermore, a highly significant correlation was found between trough CsA levels and the age of the patient, since the percentage of inhibited lymphocytes increases with age.