Paola Pedotti

Polycyclic aromatic hydrocarbon-DNA adducts in human sperm as a marker of DNA damage and infertility.

Severe DNA damage, which might prevent egg fertilization or the development of the embryo, could be a cause of infertility. In order to assess whether polycyclic aromatic hydrocarbon (PAH)-DNA adducts are an early marker of sperm genotoxicity and infertility, we studied 205 men consecutively recruited from 1 January to 30 May 2001 through the Infertility Clinic of the University of Milan (Italy), with morphological abnormalities in the sperm. No known causes of infertility were present in their female partners. Sperm were collected after 3-5 days of abstinence, fixed on polylysine slides, and frozen at -20 degrees C. PAH-DNA adducts were measured by immunofluorescence using a polyclonal antiserum. A questionnaire was filled out at the time of the visit, with demographic information, smoking and drinking habits, and occupational history. Data on PAH-DNA adducts were available for 182 men. The mean age of the subjects was 35.5+/-5.0 years; 38.6% of them were current smokers. PAH-DNA adducts were negatively correlated with the percentage of physiologic forms (r=-0.18; P=0.016) and with abnormalities of the neck of the sperm cell (r=-0.21; P>/=0.009), while they were positively correlated with morphological abnormalities of the head (r=0.30; P>0.0001). Occupational exposure to PAH, but not smoking, was significantly associated with higher levels of PAH-DNA adducts. A significant negative association was observed between daily alcohol consumption and PAH-DNA adducts in sperm (P=0.01). PAH-DNA adducts levels were significantly higher in infertile versus fertile men (P=0.04). These results suggest a role for DNA damage in infertility.

Clinical and hematologic features of 300 patients affected by hereditary spherocytosis grouped according to the type of the membrane protein defect

The molecular basis of hereditary spherocytosis is highly heterogeneous, involving the genes encoding for spectrin, ankyrin, band 3 and protein 4.2. The findings of this retrospective study show that splenectomy corrected anemia in patients with all molecular subtypes of hereditary spherocytosis. Thus, the definition of the red cell membrane defect in hereditary spherocytosis has no major clinical implications, but may be useful for a differential diagnosis from other hematologic disorders that mimic this hemolytic anemia. See related perspective article on page 1283. Background Hereditary spherocytosis is a very heterogeneous form of hemolytic anemia. The aim of this study was to relate the type of molecular defect with clinical and hematologic features and response to splenectomy using information from a large database of patients. Design and Methods Data from 300 consecutive patients with hereditary spherocytosis, grouped according to the results of sodium dodecyl sulphate-polyacrylamide gel electrophoresis, were analyzed and the sensitivity of red cell osmotic fragility tests was compared in various subsets of patients. Results Band 3 and spectrin deficiencies were the most common protein abnormalities (54% and 31%, respectively); 11% of cases were not classified by the electrophoretic analysis. Spectrin deficiency was more frequently diagnosed in childhood and band 3 deficiency in adulthood. Hemoglobin concentration was slightly lower, spherocyte number and hemolysis markers higher in spectrin deficiency than in band 3 deficiency. The sensitivity of the osmotic fragility tests ranged from 48% to 95%, and was independent of the type and amount of the membrane defect. The association of the acidified glycerol lysis test and the NaCl test on incubated blood reached a sensitivity of 99%. Splenectomy corrected the anemia in patients with all subtypes of hereditary spherocytosis although spectrin-deficient patients still showed increased reticulocyte numbers and levels of unconjugated bilirubin. Splenectomy allowed the identification of the membrane defect in all the previously unclassified patients, most of whom had spectrin and/or ankyrin deficiency. Conclusions The definition of the red cell membrane defect in hereditary spherocytosis has no major clinical implications, but may be useful for a differential diagnosis from other hematologic disorders that mimic this hemolytic anemia.

Split and whole liver transplantation outcomes: A comparative cohort study

A specific split liver transplantation (SLT) program has been pursued in the North Italian Transplant program (NITp) since November 1997. After 5 yr, 1,449 liver transplants were performed in 7 transplant centers, using 1,304 cadaveric donors. Whole liver transplantation (WLT) and SLT were performed in 1,126 and 323 cases, respectively. SLTs were performed in situ as 147 left lateral segments (LLS), 154 right trisegment liver (RTL) grafts, and 22 modified split livers (MSL), used for couples of adult recipients. After a median posttransplant follow‐up of 22 months, SLTs achieved a 3‐yr patient and graft survival not significantly different from the entire series of transplants (79.4 and 72.2% vs. 80.6 and 74.9%, respectively). Recipients receiving a WLT or a LLS showed significantly better outcomes than patients receiving RTL and MSL (P < 0.03 for patients and P < 0.04 for graft survival). At the multivariate analysis, donor age of >60 yr, RTL transplant, <50 annual transplants volume, urgent transplantation (United Network for Organ Sharing (UNOS) status I and IIA), ischemia time of >7 hours, and retransplantation were factors independently related to graft failure and to significantly worst patient survival. Right grafts procured from RTL and either split procured as MSL had a similar outcome of marginal whole livers. In conclusion, in 5 yr, the increased number of pediatric transplants due to split liver donation reduced to 3% the in‐list children mortality, and a decrease in the adult patient dropout rate from 27.2 to 16.2% was observed. Such results justify a more widespread adoption of SLT protocols, organizational difficulties not being a limit for the application of such technique. Liver Transpl 12:402–410, 2006.

Incidence of cancer after kidney transplant: Results from the north italy transplant program

Background. Patients undergoing kidney transplantation demonstrate a higher risk of developing cancer as the result of immunosuppressive treatment and concurrent infections. Methods. The incidence of cancer in a cohort of patients who underwent kidney transplantation between 1990 and 2000, and who survived the acute phase (10 days), was analyzed as part of the North Italy Transplant program. Results. A total of 3,521 patients underwent transplantation during a 10-year period in 10 of 13 participating centers; the length of follow-up after kidney transplant was 67.7±36.0 months. During the follow-up, 172 patients developed cancer (39 with Kaposi sarcoma, 38 with lymphoproliferative diseases, and 95 with carcinomas [17 kidney, 11 non-basal cell carcinoma of the skin, 10 colorectal, 8 breast, 7 gastric, 7 lung, 6 bladder, and 3 mesothelioma]). The average time to cancer development after transplant was 40.1±33.4 months (range 0–134 months). Twenty-four patients developed cancer within 6 months from the transplant (10 with carcinomas, 7 with Kaposi sarcoma, and 7 with lymphoproliferative diseases). Three patients demonstrated a second primary cancer. The average cancer incidence was 4.9%. The incidence of cancer was 0.01 per year. Independent determinants of cancer development were age, gender, and immunosuppressive protocol including induction. Ten-year mortality was significantly higher in patients with cancer (33.1%) than among patients without cancer (5.3%). The relative risk of death in subjects with cancer was 5.5 (confidence interval 4.1–7.4). Conclusions. These preliminary data underline the importance of long-term surveillance of transplant recipients, choice of immunosuppressive treatment, and careful donor selection.

Mini-review of studies on the effect of smoking and drinking habits on semen parameters

This article aims at evaluating the impact of cigarette smoking and alcohol consumption on different semen parameters (volume, concentration, motility, morphology, total sperm count and viability), by reviewing all the published literature on smoking, alcohol and sperm morphology. The studies on smoking and sperm abnormalities show a limited effect of smoking on conventional sperm parameters. The data on alcohol are more sparse, and show an apparent protective effect of moderate alcohol drinking on sperm parameters, probably due to the antioxidant effect of some alcoholic beverages. Unfortunately, none of the studies that considered both alcohol and smoking habits conducted the analysis of the joint effect of the two exposure factors. More standardized laboratory assays and increased sample size studies involving subjects with various level of exposure to alcohol and smoking are needed to better establish the role of these factors in male infertility.

A comparative prospective study of two available solutions for kidney and liver preservation

Background. Viaspan (University of Wisconsin [UW]) solution is the gold standard for abdominal organ preservation. Celsior (CEL) is an extracellular-type, low-potassium, low-viscosity solution, initially used for heart and lung preservation. We have performed a prospective multicenter study to compare the role of these cold-storage solutions on kidney and liver recovery after transplantation. Patients and Methods. From March 15, 2000 to December 31, 2001, 441 (172 CEL and 269 UW) renal transplants (RT) and 175 (79 CEL and 96 UW) liver transplants (LT) were included in the study. Results. Perfusate volume used was significantly lower in the UW group, being 4,732±796 mL versus 5,826±834 mL in the CEL group (P <0.001). In LT, median total bilirubin serum levels were significantly higher at 5 and 7 posttransplant days in the UW group (90.6 and 92.3 μmol/L, respectively) as compared with CEL (51.3 and 63.4 μmol/L, respectively). After LT, primary nonfunction (PNF) rates in the CEL and UW groups were 3.8% and 4.2% (P =NS) respectively, with 1-year graft and patient survival being 83.3% versus 85.4% (P =NS) and 89.9% versus 90.6% (P =NS). After RT, delayed graft function (DGF) rates were 23.2% and 22.7% (P =NS), respectively; PNF rates were 1.9% and 1.7% (P =NS) respectively, with 1-year graft and patient survival being 92.3% versus 94.2% (P =NS) and 99.4% versus 97.7% (P =NS). Conclusions. CEL solution was shown to be as effective as UW in both liver and kidney preservation. In LT patients, biliary function recovery is significantly better in the CEL group. CEL solution represents an efficacious option in multiorgan harvesting.

Association of metabolic gene polymorphisms with tobacco consumption in healthy controls.

Polymorphisms in genes that encode for metabolic enzymes have been associated with variations in enzyme activity between individuals. Such variations could be associated with differences in individual exposure to carcinogens that are metabolized by these genes. In this study, we examine the association between polymorphisms in several metabolic genes and the consumption of tobacco in a large sample of healthy individuals. The database of the International Collaborative Study on Genetic Susceptibility to Environmental Carcinogens was used. All the individuals who were controls from the case‐control studies included in the data set with information on smoking habits and on genetic polymorphisms were selected (n = 20,938). Sufficient information was available on the following genes that are involved in the metabolism of tobacco smoke constituents: CYP1A1, GSTM1, GSTT1, NAT2 and GSTP1. None of the tested genes was clearly associated with smoking behavior. Information on smoking dose, available for a subset of subjects, showed no effect of metabolic gene polymorphisms on the amount of smoking. No association between polymorphisms in the genes studied and tobacco consumption was observed; therefore, no effect of these genes on smoking behavior should be expected.

A common CYP1B1 polymorphism is associated with 2-OHE1/16-OHE1 urinary estrone ratio.

Abstract Cytochrome P450 (CYP) is a multigene family of enzymes involved in important life functions; some of these genes are inducible and are implicated in the oxidative metabolic activation and detoxification of many endogenous and exogenous compounds. CYP1B1 codes for an enzyme that catalyses the production of a 2- and 4-hydroxyl group in estrone and estradiol, while CYP1A1 catalyzes the 2-hydroxylation of estradiol in endometrium. The two genes were evaluated in a cohort of 150 subjects: African-American women had significantly lower 2-hydroxyl estrone/16-hydroxyl estrone (2-OHE1/16-OHE1) urinary metabolite ratios than Caucasian women (2.06±1.05 vs. 1.43±0.56; p=0.0002). A common polymorphism in the CYP1B1 gene (leucine to valineat codon 432) was associated with changes in urinary estrogen levels: both Caucasian and African-American women carrying the variant allele showed higher urinary metabolite ratios than women with the wild-type allele. No effect of the CYP1A1 MspI was observed. The 4-OHE1/2-OHE1 ratio was lower in subjects carrying the variant allele (Leu). The percentage change in 2-OHE1/16-OHE1 urinary ratio after indole treatment was significant in both Caucasian and African-American women carrying the wild-type CYP1B1 genotype, although it was more evident in African-Americans than in Caucasians. These results suggest that the Leu/Val CYP1B1 polymorphism may modify estradiol metabolism.

Epidemiologic study on the origin of cancer after kidney transplantation.

Background. Subjects who underwent solid organ transplantation are at higher risk for a wide variety of cancers. Methods. The authors investigated the origin of cancer in a cohort of 2,526 patients followed up for 60.7±35.6 months after kidney transplantation between 1990 and 2000 in seven transplant centers. Results. One hundred four of them developed cancer. All subjects who developed solid cancer within 6 months after transplantation (n=10) and a group of subjects who developed solid cancer after 6 months posttransplant (n=10) were selected. Short tandem repeat analysis was performed on paraffin-embedded biopsy specimens of tumors and on both donor and recipient pretransplant peripheral blood. Biologic material was obtained in 17 of the 20 selected patients (85.0%). The analysis showed that 16 of 17 tumors were genetically identical to the recipient. Conclusions. The authors’ results suggest that donor transmission of solid cancer is an unlikely event in their population.

Meta-Analysis Suggests Association of L-myc EcoRI Polymorphism with Cancer Prognosis

The L-myc EcoRI polymorphism is a noncoding variation in the second intron of the L-myc gene, resulting in S and L alleles. Individuals carrying the S allele tend to have poor prognosis and increased risk of several tumor types, although controversial results have been reported. A meta-analysis of 36 studies on L-myc EcoRI genotyping, including 3563 patients with different types of cancer and 2953 controls, was performed. In lung cancer patients the S/S genotype was significantly associated with lymph node metastasis [odds ratio (OR), 2.8; 95% confidence interval (CI), 1.8–4.3], distant metastasis (OR, 4.7; 95% CI, 2.4–9.2), and stage (OR, 2.3; 95% CI, 1.2–4.4). No association was observed between the S/S genotype and cancer (OR, 1.1; 95% CI, 0.8–1.4). In patients with other cancers, the S/S genotype was significantly associated with tumor recurrence (OR, 2.8; 95% CI, 1.4–6.0), whereas no significant association was seen for the other prognostic parameters. When all types of cancer were examined together, the S/S genotype was associated with lymph node metastasis (OR, 2.3; 95% CI, 1.6–3.3), distant metastasis (OR, 2.9; 95% CI, 1.8–4.6), clinical stage (OR, 1.8; 95% CI, 1.2–2.9), and cancer risk (OR, 1.25; 95% CI, 1.07–1.45). The meta-analysis suggests that the L-myc EcoRI polymorphism is a marker of tumor prognosis in lung cancer and possibly in other types of cancer.