Hyperthermic isolated limb perfusion with cis-diamminedichloro-platinum. II. An experimental study in dogs with a balloon-occlusion technique for repeated high-dose treatment.

Isolated organ perfusion is attractive for regional high-dose chemotherapy because of its advantage to reduce whole body toxicity. Intraoperative hyperthermic isolated perfusion procedures involving a heart-lung machine have been developed, but repeated treatments carry a high risk of vessel and tissue damage. Therefore, a study of isolated hyperthermic limb perfusion in four dogs was conducted using a balloon-occlusion technique including a hyperthermia unit, two low-flow rotary pumps, a bubble oxygenator, and two polyurethane balloon catheters. After 15 min infusion of cisplatinum the concentrations of serum platinum (Pt) in the isolated limb and in the whole body were measured by atomic absorption spectroscopy. Regional exposure to Pt was more than 10-fold higher than systemic exposure. After 60 min isolated limb perfusion, the area under the curve (AUC) of Pt concentrations in the isolated limb showed values between 767.4 and 1055.6 micrograms/l x 60 min, whereas in the whole body values between 59.8 and 75.9 micrograms/l x 60 min were obtained. Repeated isolated limb perfusions with the balloon-occlusion technique were performed in three dogs without systemic side effects. This model of regional chemotherapy may be useful for preoperative chemotherapy in malignant tumors of the limbs.

Diet and 20-y mortality in two rural population groups of middle-aged men in Italy.

The relationships between individual diet, measured in 1965 on the two Italian rural cohorts of the Seven Countries Study on Cardiovascular Disease, and subsequent mortality from all and specific causes of death in 20 y are studied. The analysis covers 1536 men aged 45-64 y at entry to the study. By using a cluster analysis technique, individuals are aggregated into four groups so that the elements within a group have a higher degree of similarity in dietary nutrients than between groups. Impressive differences in death rates between groups are found especially at the 10- and 15-y anniversaries. The relative risk between the least and the most favored group in 15-y mortality from coronary heart disease is 4.7; in 10 y the relative risk for cancer mortality is 2.9 and for liver cirrhosis approximately 4.

Serum thiocyanate levels as an objective measure of smoking habits in epidemiological studies.

The relationship between smoking habits and plasma thiocyanate levels has been evaluated in four adult samples of the general population containing men and women aged 20-84 and in a sample of 11-year-old non-smoking children of both sexes, for a total of 7577 individuals. Mean levels of plasma thiocyanate was found be approximately 20 mumol/l in children, 30 mumol/l in non-smoking adults and increasingly higher in smoking adults. The slopes of the regression equations of thiocyanate on cigarettes smoked per day range from 3.041 to 5.740, with correlation coefficients of from 0.638 to 0.809. In another occupational sample of 2802 men aged 46 to 65, where the correlation coefficient (between cigarette consumption and thiocyanate) was only 0.49, plasma thiocyanate was a better predictor of 4 year fatal events than cigarette consumption, 4 other covariates being considered in the same multivariate model.

Diet of two rural population groups of middle-aged men in Italy.

In 1965 a dietary survey with use of the dietary history method was carried out among 1536 middle-aged men from two rural areas of Italy, Crevalcore in the North and Montegiorgio in the Center, in connection with the Seven Countries Study on the epidemiology of cardiovascular diseases. In general the diet, while rather heterogeneous across individuals, was typically Mediterranean, that is rich in cereals, vegetables, fruits and vegetable oils, particularly olive oil and moderate in animal products. Intake of alcoholic beverages, mostly wine, was rather high. There were some differences between the two areas in consumption of some food-items such as milk, meat, fruits, sugar and cakes, pies and cookies. The dietary history method, as used here, was validated concurrently on a subsample by the 3-day weighing method and chemical analyses of 3-day food composites, with satisfactory results.

Determinants of all causes of death in samples of Italian middle-aged men followed up for 25 years.

A total of 1712 men aged 40 to 59 years in two rural cohorts of northern and central Italy have been followed up for 25 years after an entry examination in 1960. Forty one individual characteristics have been considered as possible predictors of death in the next 25 years. After exclusion of 55 men with life threatening diseases (cardiovascular and cancer) and of 161 men because of missing measurements, 1495 men have been analysed for relation between entry factors and subsequent death (n = 670). Twelve factors eventually emerged as powerful predictors of future death: in hierarchical order, age, blood pressure, forced expiratory volume, cigarette smoking, xanthelasma, mother life-status, arm circumference, father life-status, shoulder-pelvis ratio, vital capacity, arcus senilis, and serum cholesterol. Discrimination as provided by logistic modelling placed 19.6% of all cases in the upper decile of the estimated risk, 36.8% in the upper quintile, 2.5% in the lowest decile, and 7.1% in the lowest quintile. Out of those located in the lowest decile of risk, 11.4% died within 25 years while the corresponding percentage in the upper decile was 87.3%. Use of the Cox model yielded slightly better coefficients than logistic function.

The prediction of future health in healthy middle-aged men.

Two cohorts of men aged 49-59 at entry, representing cluster samples of two rural areas in Northern and Central Italy, for a total of 1712 subjects have been followed-up for 20 years within an epidemiological study originally designed for cardiovascular disease. After 20 years, only 41 men have been judged to have remained substancially healthy throughout the observation period, i.e. free from a number of major diseases. Univariate and multivariate analyses trying to predict the maintenance of health status showed that among 21 selected characteristics only the following one had a significant power: age, cigarette smoking (adverse effect) and vital capacity (favourable effect). A minor role was also played by the body mass index (adverse effect) and forced expiratory volume (favourable effect). Those who did not remain healthy exibited a greater increase in blood pressure and body mass index.

Prognostic significance of terminal transferase and adenosine deaminase in acute and chronic myeloid leukemia.

We have analyzed the distribution and prognostic significance of terminal deoxynucleotidyl transferase (TdT) and adenosine deaminase (ADA) in connection with conventional cytology, cytogenetics, response to therapy, and survival. The study population consisted of 78 patients with AML, 44 patients with Ph1 + CML in chronic phase, and 35 adult patients with Ph1 + CML in blastic phase, among which 5 cases presented as Ph1 + acute leukemia. Nine percent of the AML cases were positive for TdT and were characterized by a high percentage of blast cells in bone marrow, myeloid features by cytochemistry and absence of the Philadelphia chromosome. The median ADA values of the TdT+ AML cases were several times higher than those obtained for the TdT- cases. The survival calculated for the two groups of AML cases subdivided according to ADA levels was significantly longer (p less than 0.025) for the patients with low levels of ADA (less than 250 U/10(8) cells). In chronic phase of CML, TdT was absent and ADA values were increased over normal controls only in cases with early signs of transformation. In blastic phase, 31% of the 35 cases were positive for TdT, and ADA values were significantly higher (p less than 0.001) in TdT+ than TdT- cases. The survival calculated from the onset of transformation was significantly longer for the TdT+ acute phase (10.4 mo) compared to the TdT- patients (4.8 mo; p less than 0.025). Four cases presenting as Ph1 + acute leukemia were TdT+ and had elevated levels of ADA; 3 of them responded to ALL therapy, reverting to a stable phase of CML.

Active polypeptide fragments common to prokaryotic, eukaryotic, and mitochondrial DNA polymerases.

With a procedure that allows the renaturation of the DNA polymerase catalytic activity in situ after SDS-polyacrylamide gel electrophoresis, we have compared the active polypeptides present in extracts from organisms covering a wide evolutionary range from prokaryotes to eukaryotes, namely: Escherichia coli, Oryza sativa, Daucus carota , Neurospora crassa, Dictyostelium discoideum, Saccharomyces cerevisiae, Ceratitis capitata, Leucophaea maderae , Xenopus laevis, rat tissues and human lymphoblastoid cells. Two main clusters of active peptides are visible in mammalian and adult insect tissues, characterized by a mol. wt. greater than 70000 and less than 50000, respectively. High mol. wt. peptides are heterogeneous in size and correspond to active fragments of DNA polymerase alpha, whereas low mol. wt. peptides show the same migration rate as purified DNA polymerase beta and are not generated by proteolysis of the high mol. wt. cluster, In the three species of fungi studied, only high mol. wt. peptides are found. The same is true in plant cells, where no DNA polymerase beta activity is detectable and the pattern of the high mol. wt. cluster is similar to that observed in E. coli extracts (which also lack low mol. wt. peptides). Also in mitochondria from higher and lower eukaryotes only high mol. wt. species are observed, and the active band(s) range from 70000 to 145000 daltons. Our results indicate that the structure of DNA polymerase has been highly conserved during evolution so that an active fragment of mol. wt. greater than or equal to 70 000 is always found in prokaryotic enzymes and in the replicative species of eukaryotic and mitochondrial DNA polymerases; at a certain stage in evolution, another species of low mol. wt. DNA polymerase (beta or beta-like) appears.