Correlation of serum metalloproteinase levels with lung cancer metastasis and response to therapy.

Cancer cells elaborate metalloproteinases which may play a role in invasion and metastasis. The serum level of the M(r) 72,000 type IV collagenase (MMP-2) was measured in 87 lung cancer patients. Stage IV cancer levels were significantly elevated (P less than 0.0001) compared to normal sera. A significant difference (P less than 0.01) was found between enzyme levels in the presence versus the absence of distant metastasis. For 29 patients treated with combination chemotherapy, a positive relationship was noted between response failure and elevated enzyme levels. Serum metalloproteinase levels may provide information relevant to prognosis as well as treatment decisions.     

Heparin modulates proliferation and proteoglycan biosynthesis in murine mesangial cells: molecular clues for its activity in nephropathy

Glycosaminoglycan administration has favourable effects on morphologicaland functional renal abnormalities in different models. Thepossibility that exogenous glycosaminoglycans modulate glomerularmatrix synthesis was explored in both primary and SV40-MES13murine mesangial cell cultures. On both cell types, both low-molecular-weightheparin and different glycosaminoglycans showed dose-dependentinhibition of proliferation and increase of ³⁵SO^2–₄ uptake.After 36 h the cell compartment contained a spectrum of ³⁵S-moleculesof less than 200 kDa; under heparin treatment, the two main³⁵SO^2–₄ components (high and medium MW) increased by 16and 37% respectively. Susceptibility to glycosidases revealedthat heparin promotes the expression of heparan sulphate andincreases that of chondroitin sulphate. Moreover, heparin modifiesthe expression of decorin and bigly-can, involved in adhesionand fibrillogenesis, while not affecting perlecan. The extracellularmatrix modulation in renal cells, for which the sulphation typeand ratio of heparin are crucial, may thus explain the beneficialrenal effects of heparin.     

Hormonal and basement membrane markers for immunoidentification of cultured human trophoblast cells

Improvement of human chorionic villi cells in vitro culture has been obtained by supplementing the medium with 30% human fetal cord serum, instead of fetal calf serum. Expression of human chorionic gonadotropin, estrogen and progesterone receptors, laminin, laminin receptors, and type IV collagen has been studied on first subculture passages (4-7 weeks) by immunoperoxidase method. A minority of the cultured cells were positive for estrogen receptors, the majority were positive for progesterone receptors, while all cells were negative for human chorionic gonadotropin. Cultured cells showed variable positive immunostaining for basement membrane markers like laminin, and type IV collagen, and for laminin receptors. Detection of both progesterone receptors and laminin, or type IV collagen, excluded fibroblast contamination and could then be useful for quick identification of cultured trophoblast cells.     

Isolated hepatocyte transplantation for Crigler-Najjar syndrome type 1

Crigler-Najjar syndrome type 1 (CN1) is an inherited disorder characterized by the absence of hepatic uridine diphosphoglucuronate glucuronosyltransferase (UDPGT), the enzyme responsible for the conjugation and excretion of bilirubin. We performed allogenic hepatocyte transplantation (AHT) in a child with CN1, aiming to improve bilirubin glucuronidation in this condition. A 9-year-old boy with CN1 was prepared with plasmapheresis and immunosuppression with prednisolone and tacrolimus. When a graft was made available, 7.5 x 10(9) hepatocytes were isolated and infused into the portal vein percutaneously. After 2 weeks phenobarbitone was added to promote the enzymatic activity of UDPGT of the transplanted hepatocytes. Nocturnal phototherapy was continued throughout the studied period. Total bilirubin was considered a reliable marker of allogenic cell function. There was no significant variation of vital signs nor complications during the infusion. Mean +/- SD bilirubin level was 530 +/- 38 micromol/L before and 359 +/- 46 micromol/L after AHT (t-test, p < 0.001). However, the introduction of phenobarbitone was followed by a drop of tacrolimus level with increase of alanine aminotransferase (ALT) and increase of bilirubin. After standard treatment of cellular rejection bilirubin fell again but from then on it was maintained at a greater level. After discharge the patient experienced a further increase of bilirubin that returned to predischarge levels after readmission to the hospital. This was interpreted as poor compliance with phototherapy. Only partial correction of clinical jaundice and the poor tolerability to nocturnal phototherapy led the parents to refuse further hepatocyte infusions and request an orthotopic liver transplant. After 24 months the child is well, with good liver function on tacrolimus and prednisolone-based immunosuppression. Isolated AHT, though effective and safe, is not sufficient to correct CN1. Maintenance of adequate immunosuppression and family compliance are the main factors hampering the success of this procedure.     

Genetic study of red cell Esterase D polymorphism by ultrathin layer isoelectric focusing

Summary Human red cell Esterase D (EsD) was analyzed by isoelectric focusing (IEF) on ultrathin-layer polyacrylamide gel with a pH range of 5.0–6.0. Hemolysates were treated with Dithiothreitol to avoid loss of activity and change of the isozyme patterns by in vitro storage effects. In our sample of 951 unrelated persons from Veneto, seven different phenotypes were observed. The following allele frequencies were calculated: EsD¹=0.8476, EsD²=0.1336, EsD⁵=0.0178, and EsD^V=0.0010.Our gene frequencies have been compared to those found in other populations.Financially supported by the Ministry of Education of Italy     

Gc subtypes determined by ultrathin-layer isoelectric focusing. Distribution in the Veneto population (Italy)

Summary The distribution of Gc phenotypes in the population of Veneto was investigated by ultrathin-layer isoelectric focusing. In our sample (n=732) the six common phenotypes, Gc1S, 1F, 1S1F, 2, 2-1S, 2-1F and a further phenotype, Gc1S1C3, were observed and the following frequencies calculated: Gc1S=0.560792; Gc1F=0.159153; Gc2=0.277323; Gc1C3=0.002732. Our gene frequencies have been compared with those found in other populations.Finally supported by the Ministry of Education of Italy     

Biobanks for non-clinical purposes and the new law on forensic biobanks: does the Italian context protect the rights of minors?

Biobanks are an important resource for medical research. Genetic research on biological material from minors can yield valuable information that can improve our understanding of genetic-environmental interactions and the genesis and development of early onset genetic disorders. The major ethical concerns relating to biobanks concern consent, privacy, confidentiality, commercialisation, and the right to know or not to know. However, research on paediatric data raises specific governance and ethical questions with regard to consent and privacy. We have considered the Italian normative context focusing on what is mentioned in each document on the ethical and legal requirements that guarantee the rights of minors. We found out that there is no systematic reflection on the ethical and policy issues arising from the participation of minors in biobank research. Moreover, we have focused on the same aspects for the new Italian Law on the National Forensic Biobank.     

Paternity testing requested by private parties in Italy: some ethical considerations

In Italy, judicial and extrajudicial requests for paternity testing have increased in recent years. A retrospective analysis of such private extrajudicial requests received by the legal medicine unit of the Department of Environmental Medicine and Public Health of Padua University was conducted to identify problem areas most helpful in determining whether to accept private parties' requests for paternity testing. Such testing is most delicate when a presumptive father may be seeking to disown paternity and when testing is wanted without the consent of a member of the mother-child-father triangle. Tests that could establish paternity where none has been recognised are less problematic, as the child will not lose out. Legal and ethical-deontological aspects of consent, of the protection of minors and of children's and parents' need for follow-up interviews to deal with the outcome of such testing are carefully considered by the Padua University team when deciding whether to accept a request for testing. It is argued that because such issues are not dealt with by mail-order laboratories, the use of such services is inappropriate.