Improvement in fetal pulmonary hypertension and maturity after reversal of ductal constriction: prospective cohort study.

OBJECTIVES: To test the hypotheses that estimated mean pulmonary arterial pressure (MPAP) decreases and pulmonary vascular maturation, assessed by the ratio of pulmonary arterial flow acceleration time to ejection time (AT/ET ratio), increases after reversal of fetal ductus arteriosus constriction by reducing maternal intake of the causal agent (prostaglandin inhibitors, such as polyphenol-rich foods or non-steroidal anti-inflammatory drugs), and that these effects are independent of gestational age, which are inferences not yet demonstrated in the clinical setting. METHODS: This was a prospective cohort study comparing Doppler echocardiographic ductal flow dynamics, MPAP and pulmonary arterial flow AT/ET ratio in third-trimester fetuses (≥ 28 weeks' gestation) with ductus arteriosus constriction, at the time of diagnosis and after 2 weeks of reduced maternal intake of prostaglandin inhibitors either by suspending the use of pharmacological agents with potential for prostaglandin inhibition or by restricting the consumption of polyphenol-rich foods. MPAP was estimated using the Dabestani equation (MPAP = 90 - (0.62 × AT)), and pulmonary vascular maturity was assessed using the AT/ET ratio, according to reported validation studies. Student's t-test was used for comparison of variables at diagnosis with those after reversal of ductal constriction. Change in MPAP and pulmonary AT/ET ratio between the two assessments was compared with the expected change in the same gestational period in normal fetuses based on reference curves of MPAP and pulmonary AT/ET ratio constructed in normal fetuses from healthy pregnant women at 19-37 weeks' gestation, encompassing the same gestational age range as the study group (28-37 weeks). RESULTS: Seventy pregnancies with fetal ductus arteriosus constriction were included in the study. After 2 weeks of reduced maternal intake of prostaglandin inhibitors, normalization of mean systolic (change from 1.86 ± 0.34 m/s at diagnosis to 1.38 ± 0.41 m/s; P < 0.001) and diastolic (change from 0.41 ± 0.11 m/s to 0.21 ± 0.065 m/s; P < 0.001) ductal velocities and of mean pulsatility index (change from 1.99 ± 0.20 to 2.55 ± 0.42; P < 0.001) was demonstrated. MPAP decreased between the assessments (change from 66.7 ± 6.90 mmHg at diagnosis to 54.5 ± 6.70 mmHg after 2 weeks; P < 0.001) and mean pulmonary AT/ET ratio increased (change from 0.20 ± 0.06 to 0.33 ± 0.07; P < 0.001). Change in MPAP between diagnosis and after 2 weeks of reduced maternal intake of prostaglandin inhibitors was -12.2 ± 0.30 mmHg, which was 5.3-times higher than that in 305 normal fetuses over 2 weeks during the same gestational period (-2.3 ± 0.19 mmHg) (P < 0.001), and change in pulmonary AT/ET ratio between the two assessments was 0.13 ± 0.08, which was 8.7-times higher than that in normal fetuses in the same gestational period (0.015 ± 0.08) (P < 0.001). CONCLUSIONS: Resolution of fetal ductal constriction is followed by a fall in MPAP and by an increase in pulmonary vascular maturity, to a significantly greater degree than is observed in normal fetuses in the same gestational-age period. © 2021 International Society of Ultrasound in Obstetrics and Gynecology.

Increase of prostaglandin E2 in the reversal of fetal ductal constriction after polyphenol restriction.

OBJECTIVE: Anti-inflammatory substances that inhibit the synthesis of prostaglandins, such as non-steroidal anti-inflammatory drugs (NSAIDs) and polyphenol-rich foods, can cause constriction of the fetal ductus arteriosus. This study aimed to test the hypothesis that reversal of fetal ductal constriction after maternal restriction of polyphenol-rich foods, in the third trimester of pregnancy, is accompanied by increased plasma levels of prostaglandin E2 (PGE2). METHODS: This was a controlled clinical trial of women with singleton pregnancy ≥ 28 weeks undergoing fetal echocardiography. The intervention group included pregnancies with diagnosis of fetal ductal constriction and not exposed to NSAIDs. The control group consisted of third-trimester normal pregnancies. Both groups answered a food frequency questionnaire to assess the amount of total polyphenols in their diet, underwent Doppler echocardiographic examination and had blood samples collected for analysis of plasma levels of PGE2. Intervention group participants received dietary guidance to restrict the intake of polyphenol-rich foods. The assessments were repeated after 2 weeks in both groups. RESULTS: Forty normal pregnancies were assessed in the control group and 35 with fetal ductal constriction in the intervention group. Mean maternal age (26.6 years) and mean body mass index (30.12 kg/m2 ) were similar between the two groups. Intragroup analysis showed that dietary guidance reduced the median consumption of polyphenols (from 1234.82 to 21.03 mg/day, P < 0.001), increasing significantly the plasma concentration of PGE2 (from 1091.80 to 1136.98 pg/mL, P < 0.05) in the intervention group after 2 weeks. In addition, Doppler echocardiography showed reversal of fetal ductal constriction in the intervention group. No significant changes were observed in the control group. CONCLUSIONS: Dietary intervention for maternal restriction of polyphenol-rich foods in the third trimester of pregnancy is accompanied by increase in plasma levels of PGE2 and reversal of fetal ductal constriction. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Polyphenol supplementation inhibits physiological increase of prostaglandin E2 during reproductive period - A randomized clinical trial.

Anti-inflammatory property of polyphenols and their effect on the metabolism of prostaglandins is not established in healthy humans. This study aimed to evaluate the effect of polyphenol supplementation in plasma levels of prostaglandin E2 and other markers of inflammation and oxidative stress in women using contraceptives. In this randomized double-blind clinical trial, women aged 25-35 years were selected. Participants received capsules containing polyphenols or placebo, to be consumed for fifteen days. From 40 women randomized, 28 completed the study. Control group showed a significant increase in the levels of PGE2 (p=0.01) while the polyphenols group showed no change in these levels (p=0.79). There was an increase in hs-CRP (p<0.01) and F2-isoprostane (p=0.04) in the control group. The GSSG to GSH ratio significantly reduced in the polyphenols group (p=0.02). Supplementation with polyphenol capsules inhibited the increase in markers of inflammation and oxidative stress in women of childbearing age using combined hormonal contraceptives.

Enterprise Implementation of Digital Pathology: Feasibility, Challenges, and Opportunities.

Digital pathology is becoming technically possible to implement for routine pathology work. At our institution, we have been using digital pathology for second opinion intraoperative consultations for over 10 years. Herein, we describe our experience in converting to a digital pathology platform for primary pathology diagnosis. We implemented an incremental rollout for digital pathology on subspecialty benches, beginning with cases that contained small amounts of tissue (biopsy specimens). We successfully scanned over 40,000 slides through our digital pathology system. Several lessons (both challenges and opportunities) were learned through this implementation. A successful conversion to digital pathology requires pre-imaging adjustments, integrated software and post-imaging evaluations.

Reversal of fetal ductal constriction after maternal restriction of polyphenol-rich foods: an open clinical trial.

OBJECTIVE: To test the hypothesis that maternal restriction of polyphenol-rich foods (PRF), which, like non-steroidal anti-inflammatory drugs (NSAID), inhibit prostaglandin synthesis in the third trimester, reverse fetal ductal constriction (DC). STUDY DESIGN: An open clinical trial of 51 third trimester fetuses with DC with no history of NSAID intake was designed. All mothers were submitted to a food frequency questionnaire and were oriented to withdrawl PRF, being reassessed after 3 weeks. Doppler parameters were assessed before and after discontinuation of these substances. A control group of 26 third trimester normal fetuses, with no ductus arteriosus (DA) constriction, in which no dietary intervention was offered, was reviewed after 3 weeks. Student's t-test and Wilcoxon's test were used. RESULT: Mean gestational age was 32±3 weeks (28 to 37 weeks). After discontinuation of PRF (≥3 weeks), 48/51 fetuses (96%) showed complete reversal of DC, with decrease in mean ductal systolic velocity (1.74±0.20 m s(-1) to 1.31±0.34 m s(-1), P<0.001), mean diastolic velocity (0.33±0.09 m s(-1) to 0.21±0.07 m s(-1), P<0.001) and mean right to left ventricular dimension ratio (1.37±0.26 to 1.12±0.17, P<0.001) and increase in mean ductal pulsatility index (PI) (1.98±0.36 to 2.46±0.23, P<0.001). Median daily maternal consumption of PRF was 286 mg per day and decreased after orientation to 0 mg per day, P<0.001. In the control group, with GA of 32±4 w (29-37 w), there was no significant differences in median daily maternal consumption of PRF, mean ductal systolic velocitiy, diastolic velocity, PI and right ventricular to left ventricular diameter ratio (RV/LV) ratio. CONCLUSION: Reduction of maternal PRF intake during pregnancy, especially in the third trimester, is followed by complete reversal of DC (wide open DA), which may influence maternal dietary habits in late pregnancy.

Maternal consumption of polyphenol-rich foods in late pregnancy and fetal ductus arteriosus flow dynamics.

OBJECTIVE: To test the hypothesis that maternal consumption of polyphenol-rich foods during third trimester interferes with fetal ductal dynamics by inhibition of prostaglandin synthesis. STUDY DESIGN: In a prospective analysis, Doppler ductal velocities and right-to-left ventricular dimensions ratio of 102 fetuses exposed to polyphenol-rich foods (daily estimated maternal consumption >75th percentile, or 1089 mg) were compared with 41 unexposed fetuses (flavonoid ingestion <25th percentile, or 127 mg). RESULT: In the exposed fetuses, ductal velocities were higher (systolic: 0.96+/-0.23 m/s; diastolic: 0.17+/-0.05 m/s) and right-to-left ventricular ratio was higher (1.23+/-0.23) than in unexposed fetuses (systolic: 0.61+/-0.18 m/s, P<0.001; diastolic: 0.11+/-0.04 m/s, P=0.011; right-to-left ventricular ratio: 0.94+/-0.14, P<0.001). CONCLUSION: As maternal polyphenol-rich foods intake in late gestation may trigger alterations in fetal ductal dynamics, changes in perinatal dietary orientation are warranted.

[The Veneto Region's Registry of Dialysis and Transplantation: 2006-2007 report]. / Registro Veneto di Dialisi e Trapianto - Report 2006-2007.

INCIDENCE: Five hundred and sixty patients began renal replacement therapy in 2006, giving an incidence of 117.51 pmp; in 2007 there were 579 new patients, for an incidence rate of 120.01 pmp. Analysis of the incidence between 1998 and 2007 for both raw and age-standardized data (based on the 2001 census) shows a slow, gradual increase that is statistically significant. Most of the patients were between 55 and 85 years old; the modal class for males was between 65 and 70, and between 75 and 80 for females. The median age of the population beginning replacement therapy is clearly over 65 years old. The year 2000 was particularly significant because the incidence of new patients undergoing renal replacement therapy over the age of 75 definitively exceeded that of the next younger class (65-74 years old), a trend that remained constant until 2007. In 2006 and 2007, males account for 64.4% and 66.4%, respectively, of new patients, a proportion that is constant over the years. The greater incidence of males is also to be found across the other age groups and tends to be even more noticeable in the oldest age class. Incidence by province is highly variable, however, there is a constant trend within provinces during these years, since the incidence in some provinces is lower than the regional average and higher in others. After adjusting for age, there are no significant differences in the incidence between provinces: the age structure of the population accounts for the variability of the incidence of terminal uremia across the Veneto provinces. The conditions most responsible for renal insufficiency requiring replacement therapy are vascular diseases, diabetes and nephropathies of unknown origin. Although diabetic and vascular nephropathies are subject to wide fluctuations, they remain stable over the years, whereas the frequency of nephropathy of unknown origin appears to be on the rise. The first treatment for most of the patients is hemodialysis. In 2006, 436 patients (78%) were given extracorporeal dialysis as first treatment, compared to 122 patients (22%) who were given peritoneal dialysis and 2 (0.35%) who received live-donor kidney transplant. In 2007 the situation was very similar, with 435 patients treated with extracorporeal dialysis, 142 with peritoneal dialysis and 1 by a live-donor transplant. The proportion between patients treated with hemodialysis and peritoneal dialysis was constant from 1998 to 2007. The choice between hemodialysis or peritoneal dialysis as the initial treatment modality depends on many factors, ranging from clinical indications to cultural attitudes at the facility to individual patient preferences. Logistic regression of the factors influencing the choice of dialysis treatment shows that peritoneal dialysis is offered primarily to patients between the ages of 45 and 65 who do not have an underlying systemic or nephropathy of unknown origin and who do not have any comorbidities. This confirms the positive selection made with regard to these patients, widely described in the literature. Initial treatment by transplant is an exceptional event: starting from 2003, it was used in only 1 or 2 patients per year. Seventy-two percent of patients starting replacement therapy present with at least one comorbidity. Thirty-six percent of patients also present with more than one associated disease. The RVDT has been gathering data on the vascular access used for new dialysis patients since 2006. Roughly 43% of patients start treatment with an arteriovenous fistula, 38% with a temporary catheter, less than 1% with a prosthesis, 9% with a tunneled catheter, and 10% with a peritoneal catheter. Logistic regression was used to evaluate what role age, primary nephropathies and comorbidities present at the start of treatment play in determining the choice of a temporary catheter. The logistic model estimates a 29% probability of starting treatment with a temporary access. This probability decreases if the patient suffers from a familiar or hereditary nephropathy but increases if the patient has secondary glomerulonephritis or is affected by a group of various diseases (multiple myeloma or other pathologies) or if the patient suffers at the same time from cardiac insufficiency or an infection. The estimated probability of starting hemodialysis with a mature fistula is 40%, but this figure diminishes significantly in female patients, if the patient has secondary glomerulonephritis, cardiac insufficiency or infections. PREVALENCE: As of December 31, 2006, there were 4,071 patients being treated with extracorporeal or peritoneal dialysis or by kidney transplant, leading to a prevalence of 852.82 patients pmp; as of December 31, 2007, there were 4,200 patients treated, with a corresponding prevalence of 869.14 pmp. The breakdown in prevalence by age group shows that the increase in prevalence is highly significant in the top two age classes, namely, between 65 and 75 years of age and over 75, while remaining negligible in the other classes. Between 1998 and 2007, the prevalence increased by 40% in patients over 75 and increased by 20% in the class of 65-to-75 year olds. The elderly contribute a greater weight in the renal replacement therapy population, reflected in the gradual increase of the median age of the prevalent population from 1998 to 2007. During 2006 and 2007, males made up 63.99% and 64.36% of the patients, respectively. This relative frequency mirrors the findings for incidence and is constant over the years. The distribution of primary diseases is very different in the prevalent population compared to findings in the incident patients. Primary glomerulonephritis, at fourth place among incident patients, is the most frequent disease in the prevalent population (although there is a clearly downward trend over the years). The percentages of diabetes and vascular disease, on the other hand, are lower compared to what is observed in the incident population. The prevalence expressed by treatment modality pmp increased for all three types. In analyzing the annual percentage rise in prevalence, using 1998 as the baseline, the most significant figure regards transplant patients, whose prevalence increased by over 60% between 1998 and 2007. Prevalence of hemodialysis patients rose moderately by only slightly over 10%. Peritoneal dialysis shows a rather linear increase, similar to the transplant trend. Our study used longitudinal regression models to analyze factors predictive of a patient starting and continuing to undergo the same type of treatment over the years. The results show that a patient has a greater probability of being treated with hemodialysis based on several primary nephropathies, when aged > 45, and in the presence of the main comorbidities. The predictive factors for peritoneal dialysis mentioned earlier have a diametrically opposed role. The presence of comorbidities (except high blood pressure), the type of nephropathy, and age > 65 lead to a lower probability of receiving a transplant. We analyzed peritoneal dialysis failures - defined as changing over to extracorporeal dialysis for any reason (clinical, psychological or social) - and the cumulative incidence of failure, taking into account the two competing outcomes of transplant and death. The only variable associated with peritoneal dialysis failure was the presence of infections. Older patients, patients with peripheral vascular disease, and those with neoplasia are less frequently taken off peritoneal dialysis to receive a transplant, an event occurring more frequently, however, in patients with hypertension. Death is dependent on age, on the presence of peripheral vascular disease and is less frequent in hypertensives. As is the case for peritoneal dialysis, the natural history of kidney transplant can have two competing outcomes: return to dialysis and death. The risk factors associated with return to dialysis are the presence of peripheral vascular disease, hypertension and infections; risk factors associated with death include age, the presence of cerebral vascular disease and neoplasia. From 1998 to 2007, the prevalence of hepatitis C virus-antibody-positive patients decreased by almost one third. The number of antigen-positive hepatitis B patients is declining slowly, but the levels remain in any case very low. The association between the two infections is disappearing: already at very low levels in 1998, that figure was halved by 2007. MORTALITY AND SURVIVAL: The mortality of uremic patients on renal replacement therapy was calculated both as a cumulative incidence, expressed as the number of deaths over patients at risk (alive at the beginning of the study year) and as a mortality rate, expressed as the number of deaths per patients/year. The figure was constant over the years, at around 10%. The mortality of males was no different from that of females; this finding differs from what is observed in the general population where male mortality is markedly higher than that of females. The mortality rate is dependent on the age group of the patient at start of treatment and shows an upward trend that is growing exponentially. The mortality rate in hemodialysis patients remained stable at 15% between 2000 and 2007, while the mortality rate in peritoneal dialysis patients gradually decreased down to 13%. The mortality rate for transplant patients was low and constant, at under 2%. The trend for the various causes of death is stable over the years and shows that the main cause of death is cardiac, accounting for between 30% and 35%, while mortality due to vascular, neoplastic, infection or cachexia-related causes are all roughly at the same rate, between 10% and 15%. (ABSTRACT TRUNCATED)

Antioxidant activity of polyphenols from solid olive residues of c.v. Coratina.

The antioxidant profile of extracts from solid olive residue (SOR) of c.v. Coratina, a cultivar widely diffused in the south of Italy, using both cell-free and cell-based experimental models, was investigated. A total hydroalcoholic extract (polyphenols content 19.7%) and a purified extract (Oleaselecttrade mark) (polyphenols content 35.1%) were tested for their ability to quench the stable free radical DPPH, the peroxyl radicals (ORAC assay), by monitoring the loss in fluorescence of R-phycoerythrin induced by the peroxyl radical generator AAPH and their ability to inhibit the cumene hydroperoxide-induced lysis of rat red blood cells (RBC). The total hydroalcoholic extract showed IC(50) 26.96+/-1.53 microg/ml in the DPPH assay, that 10 microg/ml were equivalent to 2.11+/-0.12 microg/ml Trolox (ORAC assay) and IC(50) 1.7+/-0.20 microg/ml in the RBC hemolysis. The Oleaselect extract was 4 to 5 folds more active than the hydroalcoholic extract in all the experimental models, with IC(50) values of 7.36+/-0.38 microg/ml in the DPPH test and of 0.38+/-0.03 microg/ml in RBC; the antioxidant activity in the ORAC assay was slightly greater than that of Trolox (10 microg/ml equivalent to 11.45+/-0.40 microg/ml). The scavenging effect of the extract in the ORAC assay was compared to that of verbascoside (the main polyphenol component) and of caffeic acid (the basic constituent of verbascoside): the results indicate that caffeic acid (10 microg/ml equivalent to 35.70+/-2.95 microg/ml Trolox) is more potent than verbascoside (10 microg/ml equivalent to 15.42+/-1.21 microg/ml Trolox) in entrapping peroxyl radicals. Finally the antioxidant activity of the Oleaselect extract was confirmed in human umbilical endothelial cells (EC) exposed to the site-specific peroxyl radical inducer AAPH, where a massive lipid peroxidation process (marker the fluorescence probe BODIPY) takes place, paralleled by a marked loss of cell viability (calcein assay). The purified extract (1-20 microg/ml) pre-incubated with EC for 1 h dose-dependently inhibited both the lipid-peroxidation damage and cell death. Taking into account the total polyphenol content, these results clearly indicate a greater antioxidant activity for the purified extract, due to a cooperative antioxidant interaction among its polyphenol constituents.

Rheumatoid and psoriatic knee synovitis: clinical, grey scale, and power Doppler ultrasound assessment of the response to etanercept.

OBJECTIVE: To determine the effect of tumour necrosis factor alpha (TNFalpha) blockade with etanercept in refractory knee joint synovitis (KJS) in rheumatoid and psoriatic arthritis, by local and systemic disease activity assessment and combined grey scale and power Doppler ultrasonographic monitoring. METHODS: 27 knees affected by rheumatoid KJS (n = 12) and psoriatic KJS (n = 8) were assessed before receiving treatment and at 3 and 12 months' follow up. Time dependent clinical changes in disease activity were monitored by C reactive protein, erythrocyte sedimentation rate (ESR), global health status (GHS), and Ritchie (RAI) and knee joint articular (KJAI) indices; synovial changes were monitored by ultrasonographic and power Doppler indices for grey scale synovial thickening and for distinct intrasynovial vessel power Doppler flow configurations (fluid/synovium interface (F/SI-PD) and pannus/cartilage interface (P/CI-PD)). Interobserver and intraobserver variability of grey scale and power Doppler ultrasonographic was evaluated. Response to treatment was assessed by analysis of variance for repeated measures on clinical and ultrasonographic variables. RESULTS: Rapid (3 months) reduction in F/SI-PD flow (p<0.001), parallel to reductions of C reactive protein (p<0.05), ESR (p<0.001), KJAI (p<0.002), RAI, and GHS (p<0.001), was sustained at 12 months when it was accompanied by reduction in both synovial thickening and P/CI-PD flow (p<0.001). No differences (ANOVA) were noted at baseline or at 12 months in clinical and ultrasonographic variables between either the rheumatoid or the psoriatic KJS groups. CONCLUSION: Grey scale and power Doppler ultrasonography are reliable measures of long term change in rheumatoid and psoriatic KJS disease activity in response to anti-TNFalpha treatment with etanercept.

Alternative parameters for echocardiographic assessment of fetal diastolic function

Alternative methods to assess ventricular diastolic function in the fetus are proposed. Fetal myocardial hypertrophy in maternal diabetes was used as a model of decreased left ventricular compliance (LVC), and fetal respiratory movements as a model of increased LVC. Comparison of three groups of fetuses showed that, in 10 fetuses of diabetic mothers (FDM) with septal hypertrophy (SH), the mean excursion index of the septum primum (EISP) (ratio between the linear excursion of the flap valve and the left atrial diameter) was 0.36 ± 0.09, in 8 FDM without SH it was 0.51 ± 0.09 (P = 0.001), and in the 8 normal control fetuses (NCF) it was 0.49 ± 0.12 (P = 0.003). In another study, 28 fetuses in apnea had a mean EISP of 0.39 ± 0.05 which increased to 0.57 ± 0.07 during respiration (P < 0.001). These two studies showed that the mobility of the septum primum was reduced when LVC was decreased and was increased when LVC was enhanced. Mean pulmonary vein pulsatility was higher in 14 FDM (1.83 ± 1.21) than in 26 NCF (1.02 ± 0.31; P = 0.02). In the same fetuses, mean left atrial shortening was decreased (0.40 ± 0.11) in relation to NCF (0.51 ± 0.09; P = 0.011). These results suggest that FDM may have a higher preload than normal controls, probably as a result of increased myocardial mass and LV hypertrophy. Prenatal assessment of LV diastolic function by fetal echocardiography should include analysis of septum primum mobility, pulmonary vein pulsatility, and left atrial shortening.

Effect of epoetin on HO-1 mRNA level and plasma antioxidants in hemodialysis patients.

OBJECTIVE: Patients with renal failure and undergoing hemo- (HD) or peritoneal dialysis are under oxidative stress which is thought to contribute to the long-term complications noted in this patient population. One effect of HD-induced oxidative stress is via red blood cell (RBC) membrane lipid peroxidation leading to RBC destruction and anemia. Interaction of this oxidative stress with epoetin (EPO) treatment to increase RBC number and Hb concentration remains unexplored. PATIENTS AND METHODS: This preliminary study used RT-PCR as well as colorimetric based assay approaches to evaluate the effect of EPO-alpha treatment on markers of oxidative stress in hemodialysis patients. Eighteen patients (12 males, 6 females, age range 45 - 68), were treated with EPO-alpha (Eprex) 50 UI/kg thrice weekly over an 8-month study period. Monocytes were isolated at baseline, then monthly thereafter, monocyte heme-oxygenase-1 (HO-1) and plasma Hb and antioxidant power (AOP) were determined. RESULTS AND CONCLUSIONS: Treatment with EPO increased Hb (9.4 +/- 0.7 g/dl to 10.9 +/- 0.5, mean +/- SD p < 0.001). In addition, both monocyte HO-1 mRNA (0.34 +/- 0.08 vs. 0.59 +/- 0.02 d.u. p < 0.001) and plasma AOP (1,379.8 +/- 175 micromol/l to 1,624 +/- 170, p < 0.04) increased. While AOP changes showed no correlation with other indices, increases in HO-1 and Hb were positively correlated using 2 different measures: delta Hb (peak Hb - baseline Hb) vs. delta HO-1 (peak HO-1 mRNA - baseline HO-1 mRNA) as well as delta Hb(5 months-baseline) vs. delta HO-1 (5 months - baseline) mRNA (r = 0.81, p < 0.001 and r = 0.76, p < 0.001; respectively). In conclusion, the increases upon EPO treatment of both HO-1 gene expression and plasma AOP as well as the significant correlation between delta Hb and delta HO-1 mRNA suggest that EPO treatment reduces oxidative stress via a combination of effects. These could potentially include effects on oxidative stress directly as well as effects on the levels and types of antioxidants present in plasma.

[Latent celiac disease in subjects with serum anti-endomysial antibodies and normal intestinal biopsy]. / Celiachia latente in soggetti con anticorpi antiendomisio positivi e biopsia intestinale normale.

OBJECTIVES: Data on the follow-up of a group of subjects with serum antiendomysial antibodies (EMA) and normal mucosal architecture at the intestinal biopsy are reported. Clinical problems concerning possible evolution of potential celiac disease (CD) towards gluten-induced histological damage are discussed. METHODS: Eleven patients belonging to high-risk groups for CD (5 with type-1 diabetes, 2 with familiarity for CD and 4 with symptoms suggesting CD) who had a normal intestinal biopsy, despite positive antiendomysial test, were followed-up. Antigliadin and antitransglutaminase antibodies (anti-tTG) and HLA genotyping were also assessed. According to clinical and serological data a second biopsy was performed in six of them. RESULTS: At the time of the first normal biopsy, all patients were positive for EMA and 5/8 for anti-tTG. Five of 6 subjects genotyped were HLA-DQ2+ or DQ8+. Six patients were rebiopsed after 1 to 4 years. Three had mucosal atrophy, 1 had mild increase of intraepithelial lymphocytes and 2 were morphologically normal. CONCLUSIONS: Subjects with antiendomysial antibodies and normal intestinal biopsy deserve clinical and serological follow-up to reduce the time of possible latency of CD. Although good predictors of progression of the disease are not still available, antiendomysial antibodies assessment and HLA genotyping may help to suggest individuals at higher risk to develop gluten-induced enteropathy. This study confirms that subjects with persistent signs of gluten sensitivity and normal biopsy should be re-examined.

Renal involvement in subjects with peripheral atherosclerosis.

BACKGROUND: Ischemic nephropathy is an important cause of renal failure in western countries. Subclinical renal function abnormalities may exist in patients with extrarenal atherosclerosis, and may precede the onset of overt ischemic nephropathy. METHODS: To assess the impact of extrarenal atherosclerosis on the kidney, we evaluated renal function in 89 subjects with differing degrees of peripheral atherosclerosis, without manifest clinical or laboratory signs of ischemic nephropathy and renovascular hypertension. All laboratory testing, ultrasonography with Doppler analysis for the localization of peripheral vascular disease (carotid and lower limb arteries), and non-invasive evaluation of renal function by radionuclide studies of renal plasma flow (MAG3 clearance) and glomerular filtration (DTPA clearance), as well as total, LDL and HDL cholesterol, and triglycerides were determined; smoking habit was recorded. By combining sonographic data on arterial tree stenosis (ATS), the subjects were grouped according to the atherosclerotic vascular damage (ATS involvement). RESULTS: Despite no change in plasma creatinine and DTPA clearance (from 91.58+/-26.53 mL/min/1.73 m2 to 93.47+/-24.82), MAG3 clearance progressively declined with the severity of vascular damage (from 244.86+/-60.60 mL/min/1.73 m2 to 173.59+/-58.74). Stepwise multiple regression analysis indicated that MAG3 clearance was best explained by ATS involvement (standardized beta coefficient -0.40; p<0.001), smoking habit (-0.34; p= 0.004), and serum LDL-cholesterol (-0.24; p<0.035). CONCLUSIONS: The renal hemodynamic profile in atherosclerotic patients might constitute functional evidence of the silent phase of ischemic renal disease. The findings suggest that renal function should be carefully assessed in patients with extrarenal atherosclerosis, particularly in those with classic cardiovascular risk factors.

Endothelial dysfunction in small resistance arteries of patients with non-insulin-dependent diabetes mellitus.

OBJECTIVE: Arterial hypertension is frequently associated with the presence of endothelial dysfunction in human subcutaneous small resistance arteries, as evaluated by responses to acetylcholine or bradykinin; however it is not known whether patients with diabetes mellitus show similar alterations. Therefore, we have investigated endothelial function in subcutaneous arteries of normotensive subjects (NT), of patients with essential hypertension (EH), of patients with non-insulin-dependent diabetes mellitus (NIDDM), as well as of patients with both essential hypertension and non-insulin-dependent diabetes mellitus (NIDDM+EH). PATIENTS AND METHODS: All subjects were submitted to a biopsy of the subcutaneous fat Small arteries were dissected and mounted on a micromyograph. The media to lumen ratio (M/L) was calculated. A concentration-response curve to acetylcholine, to bradykinin as well as to the endothelium-independent vasodilator sodium nitroprusside were performed. We also evaluated the contractile response to endothelin-1. Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) plasma levels were also measured. RESULTS: The vasodilatation to acetylcholine and bradykinin (but not to sodium nitroprusside) was significantly and similarly reduced in EH, in NIDDM, and in NIDDM+EH compared with NT. The contractile response to endothelin-1 was similarly reduced in EH, in NIDDM and in NIDDM+EH. Plasma ICAM-1 and VCAM-1 concentrations were higher in EH, NIDDM and NIDDM+EH than in NT. CONCLUSIONS: An evident endothelial dysfunction was detected in patients with NIDDM, and the simultaneous presence of EH did not seem to exert an additive effect. The contractile responses to endothelin-1 were reduced possibly as a consequence of ET(A) receptor down-regulation.

Platelet/polymorphonuclear leukocyte adhesion: a new role for SRC kinases in Mac-1 adhesive function triggered by P-selectin.

Adhesion of polymorphonuclear leukocytes (PMNLs) to activated platelets requires a P-selectin-triggered, tyrosine kinase-dependent adhesiveness of Mac-1 and is accompanied by tyrosine phosphorylation of a 110-kd protein (P-110) in PMNLs. Inhibitors of SRC tyrosine kinases were found to inhibit PMNL adhesion to activated platelets or to P-selectin expressing Chinese hamster ovary (CHO-P) cells and the tyrosine phosphorylation of P-110. Adhesion of PMNLs to activated platelets or to CHO-P cells stimulated activity of LYN and HCK. Monoclonal antibody blockade of P-selectin or beta2-integrins reduced the activation of both kinases. In PMNLs either adherent to platelets or aggregated by P-selectin-IgG chimera, Mac-1 was rapidly redistributed to the Triton X-100-insoluble cytoskeletal fraction, and large clusters of Mac-1 colocalized with patches of F-actin at the sites of cell-cell contact. In PMNLs stimulated by P-selectin-IgG chimera, SRC kinase inhibition impaired Mac-1 clustering, F-actin accumulation, and CD18 redistribution to the cytoskeleton. Disruption of the actin filament network by cytochalasin D prevented PMNL-platelet adhesion and P-selectin-induced PMNL aggregation and impaired the clustering of Mac-1. In agreement with the requirement for the beta2-integrin in the functional up-regulation of LYN and HCK, integrin blockade by monoclonal antibodies resulted in a complete inhibition of P-selectin-induced Mac-1 clustering and F-actin accumulation. Taken together, the results indicate that, after an initial P-selectin-triggered beta2-integrin interaction with the ligand, SRC kinases are activated and allow the remodeling of cytoskeleton-integrin linkages and integrin clustering that finally strengthen cell-cell adhesion. This model highlights a new role for SRC kinases in a regulatory loop by which the Mac-1 promotes its own adhesive function.

Cellular hypertrophy in subcutaneous small arteries of patients with renovascular hypertension.

Structural alterations of small arteries in patients with essential hypertension are characterized by inward eutrophic remodeling. However, small arteries in patients with secondary hypertension, as well as in experimental models of hypertension with high circulating renin, are characterized by inward hypertrophic remodeling, which is characterized by smooth muscle cell hypertrophy in animal models. The aim of our study was to determine whether remodeling of subcutaneous small arteries in patients with secondary forms of hypertension is associated with smooth muscle cell hypertrophy and/or alterations in the elastic modulus of the vessel wall. Fifteen patients with renovascular hypertension, 9 with primary aldosteronism, and 13 with essential hypertension and 9 normotensive subjects were included in the study. A biopsy of subcutaneous fat was taken from all subjects. Small arteries were dissected, and morphology was determined on a micromyograph. Unbiased estimates of cell volume and number were made in fixed material. From the resting tension-internal circumference relation of the small arteries, the incremental elastic modulus was calculated and plotted as a function of wall stress. Blood pressure was greater in patients with essential hypertension, renovascular hypertension, or primary aldosteronism than in normotensive subjects, but no significant difference was observed among the 3 groups of hypertensive patients. The media/lumen ratio, the medial cross-sectional area, and the smooth muscle cell volume were significantly greater in patients with renovascular hypertension than in normotensive subjects and patients with essential hypertension. No difference in cell number or in the elastic properties was observed among the 4 groups of subjects. In conclusion, our data demonstrate for the first time that a pronounced activation of the renin-angiotensin-aldosterone system is associated with vascular smooth muscle cell hypertrophy in human hypertension in a manner similar to that found in animal models.

Altered tissue electric properties in lung cancer patients as detected by bioelectric impedance vector analysis.

Modifications of body composition are frequent in cancer patients. Bioelectric impedance analysis can specifically detect changes in tissue electric properties, which may be associated with outcome. We evaluated the distribution of the impedance vectors from 63 adult male patients with lung cancer, stages IIIB (33 patients) and IV (30 patients), in supportive therapy. Body weight change over the previous 6 m.o. was the same in both groups (stable/increased 36% and decreased in 62%). Patients were compared with 56 healthy subjects matched for gender, age, and body mass index (25 kg/m2). Impedance measurements (standard tetrapolar electrode placement on the hand and foot) were made with 50-kHz alternating currents. The resistance and reactance of the vector components were standardized by the height of the subjects and were plotted as resistance/reactance graphs. The impedance vector distribution was the same in patients with either stage IIIB or IV cancer. The mean vector position differed significantly between cancer patients and control subjects (Hotelling T2 test, P < 0.01) because of a reduced reactance component (i.e., a smaller phase angle) with preserved resistance component in both cancer groups. Patients with a phase angle smaller than 4.5 degrees had a significantly shorter, i.e., 18 m.o., survival. Body weight loss was not significantly associated with survival. In conclusion, impedance vectors from lung cancer patients were characterized by a reduced reactance component. The altered tissue electric properties were more predictive than weight loss of prognosis.

Bone metabolism in orthotopic liver transplantation: a prospective study.

Bone mineral density (BMD) and mineral metabolism were assessed in 54 patients with end-stage liver disease who were evaluated for orthotopic liver transplantation (OLT) and assessed 3, 6, and 12 months after surgery in 26 patients who underwent OLT. Serum and urinary electrolyte and mineral levels, serum liver function test results, and parathyroid hormone (PTH), osteocalcin (BGP), 25-hydroxyvitamin D, and urinary hydroxyproline levels were assessed. BMD of the lumbar spine was measured at baseline and 3, 6, and 12 months after OLT. At baseline, 40.7% of patients had BMD below the fracture threshold (0.800 g/cm2). Using multiple stepwise regression analysis, we found that BMD was significantly (P < .0001) affected by age, serum creatinine level, and PTH level but not by indices of cholestasis or liver function. In the patients who underwent OLT, a 1.4% reduction (P < .006) was observed in BMD 3 months after OLT. Thereafter, BMD returned to pretransplant values. A significant increase in serum BGP was observed after 6 (P < .02) and 12 (P < . 005) months. PTH levels increased progressively 3 (P < .02), 6 (P < . 001), and 12 (P < .0001) months after OLT. This increase did not seem to be caused by cyclosporine-induced nephropathy. It was concluded that osteopenia is a major complication in hepatic cirrhosis, regardless of its causes. The increase in serum BGP levels 6 and 12 months after OLT indicates metabolic activation of osteoblasts. The increase in PTH levels after OLT warrants further investigation.

Isolation and characterization of a digoxin-like immunoreactive substance from human urine by affinity chromatography.

A series of observations has suggested that one or more digoxin-like immunoreactive substances (DLIS) in biological fluids is able to cross-react with the antidigoxin antibody. Whether this substance is the endogenous inhibitor of Na+/K+ ATPase has not been well established. The aim of this study was to identify and characterize DLIS from human urine. Treated urine from healthy men was run on an affinity chromatography column at a flow rate of 1 mL/min in which the ligand was an antibody (antiserum) to digoxin. Eluates from affinity chromatography were applied onto analytical reversed-phase HPLC. The active material was eluted with a linear gradient of acetonitrile (from 350 to 650 mL/L) and water. A second step in HPLC was carried out isocratically with 280 mL/L acetonitrile in water. We found a single peak showing cross-reactivity with antidigoxin antibody as measured by RIA. It showed the same retention time as that of a digoxin calibrator. This highly purified substance is able to displace [3H]ouabain from dog kidney-derived Na+/K+ ATPase, to inhibit Na+/K+ ATPase activity as measured by the 86Rb+ uptake in red blood cells and by coupled enzyme assay. Our results are consistent with the hypothesis that DLIS, as isolated by this particular digoxin antibody, is a single substance and an inhibitor of Na+/K+ ATPase.

Recurrence after resection for ductal adenocarcinoma of the pancreas.

We analyzed the pattern of failure and clinicopathologic factors influencing the disease-free survival of 78 patients who died after macroscopic curative resection for pancreatic cancer. Local recurrence was a component of failure in 56 patients (71.8%) and hepatic recurrence in 48 (61.5%), both accounting for 97% of the total recurrence rate. About 95% of recurrences occurred by 24 months after operation. Median disease-free survival time was 8 months, and cumulative 1-, 3-, and 5-year actuarial disease-free survival rates were 66%, 7%, and 3%, respectively. Multivariate analysis showed that tumor grade (p = 0.04), microscopic radicality of resection (p = 0.04), lymph node status (p = 0.01), and size of the tumor (p = 0.005) were independent predictors of disease-free survival. Patterns of failure and disease-free survival were not statistically influenced by the type of surgical procedure performed. Median survival time from the detection of recurrence until death was 7 months for local recurrence versus 3 months for hepatic or local plus hepatic recurrence (p < 0.05). From our experience and the data collected from the literature, it appears that surgery alone is an inadequate treatment for cure in patients with pancreatic carcinoma. Effective adjuvant therapies are needed to improve locoregional control of pancreatic cancer after surgical resection.