Phase I trial of intravenous infusion of ex-vivo-activated autologous blood-derived macrophages in patients with non-small-cell lung cancer: Toxicity and immunomodulatory effects

Summary The purpose of this phase I study was to evaluate the toxicity and biological activity of autologous blood-derived macrophages activated ex-vivo with recombinant human interferon (rhuIFN) [monokine-activated killer (MAK) cells] and administered intravenously to 11 lung cancer patients once a week for 6 consecutive weeks. Peripheral blood monocytes were collected by leukapheresis and then purified by counterflow elutriation. The MAK cells were generated by culturing the purified monocytes in Teflon bags for 7 days and adding rhuIFN to the cultured cells for the last 18 h. These MAK cells expressed differentiation-associated surface antigen MAX1, and were cytotoxic in vitro against tumour cell line U937. The MAK cells were infused at dose levels from 1 × 10⁷ to 5 × 10⁸ on an intrapatient dose-escalating schedule. No severe adverse side-effects occurred. Toxicity was mild to moderate [primarly fever (75%) and chills (32%)], non-dose-dependent, and non-cumulative. No consistent change in haemostatic function, or liver or renal function was observed. Dose-limiting toxicity was not reached at 5 × 10⁸ cells (optimal dose reproduced for each patient). The maximum tolerated dose was not determined. The immunomodulatory activity of i.v. infused MAK cells was demonstrated both in vivo by significant increases in granulocyte count and neopterin level in the patients' peripheral blood postinfusion and in vitro by secretory products (IL-1. TNF, neopterin, and thromboplastin-like substance) in the culture supernatants. The in vivo traffic patterns of autologous MAK cells labelled ex-vivo with¹¹¹In oxine were studied in 7 patients. Gamma imaging showed an immediate but transient lung uptake (<24 h), and a progressive uptake of radioactivity in the liver and spleen was seen from 6 h to 72 h post-infusion. Our results indicate that the preparation of high numbers of autologous, blood-derived MAK cells is a feasible procedure, and their transfusion is safe for patients. This immunotherapeutic approach seems to be encouraging from the point of view of establishing an adjuvant therapeutic modality in cancer patients with minimal residual disease.This work was supported in part by a grant 6911 from the Association pour la Recherche contre le Cancer (ARC), grants from the Ligue Nationale contre le cancer and the Ligues Regionales (Bas-Rhin, Haut-Rhin) contre le cancer, and contract 891013 from the Institut National pour la Santé et la Recherche Médicale (INSERM), France     

Effects of extremely low frequency (ELF) electric fields on cell growth and DNA repair in human skin fibroblasts

A number of physical and chemical agents in the environment have been studied for their ability to induce or alter DNA repair mechanisms in human cells. We have investigated the effects of 60 Hz, 1000 V/cm electric fields on DNA repair in normal human fibroblasts in vitro. An examination was done on the ability of electric fields suspected to cause damage which could be repaired by thymine dimer excision and measurable by the bromodeoxyuridine photolysis assay. The thymine dimer assay with enzyme-sensitive site analysis was used to measure the cells' capacity for removing ultraviolet light (u.v.)-induced pyrimidine dimers; during exposure to electric field 24 hr before u.v. irradiation; 24 hr after u.v. irradiation; and up to 48 hr continuously after u.v. irradiation. Cell growth and cell survival following electric field exposure were also studied. Within the limits of these experiments, it was found that exposure to such electric fields did not alter cell growth or survival, and no DNA repair or alteration in DNA excision repair capacity was observed as compared with unexposed control cultures.     

Transformation of the basidiomycete, Schizophyllum commune

Summary Protoplasts of aSchizophyllum commune tryptophan auxotroph (trp1), deficient in indole-3-glycerol phosphate synthetase (IGPS), were transformed to trp⁺ with plasmid DNA containing the SchizophyllumTRP1 sequence. Efficiencies up to 30 transformants per microgram of plasmid DNA were obtained. Southern blots reveal that the transforming DNA is integrated in chromosomal DNA. The trp⁺ phenotype of transformants is stable in meiosis and mitosis. Transformants possess IGPS activity comparable to wild-type cells.     

Amino Acid Analysis Demonstrates that Increased Plasma Free Tryptophan Causes the Increase of Brain Tryptophan During Exercise in the Rat

Rats were trained to run on a horizontal treadmill for 2 h at 20 m/min. This activity considerably increased plasma free tryptophan (TRP) (+70%) but did not alter plasma total TRP levels and had little or no effect on plasma concentrations of the other large neutral amino acids (LNAAs) that compete with TRP for entry into the brain. Brain TRP levels increased by 80%. The only other brain LNAA to be affected by exercise was threonine, which rose moderately. The results indicate that increased plasma free TRP was specifically responsible for the increase of brain TRP after 2 h of exercise. Brain lysine was also increased whereas glycine, alanine, and gamma-aminobutyric acid were decreased. The differences between the present findings and those previously obtained following 2 h immobilization stress are discussed.     

Adaptation, Readaptation and Synthesis of Hydrogenases in Scenedesmus obliquus

Scenedesmus cells reach full hydrogen activity after 2.5 h ofanaerobic adaptation. Exposure to oxygen inactivates the hydrogenaseimmediately. Readaptation occurs with the same kinetics as primaryadaptation. The fast activation and reactivation as well asthe insensibility to cycloheximide suggest that hydrogenase,inactive or activated, is present in the chloroplast all thetime. The activation and even more the readaptation are sensitiveto chloramphenicol. Thus, we propose that hydrogenase itselfor an activating protein in the chloroplast have a rather fastturnover. ¹ Present address: Pharathiar University, Dept. of Botany, Coimbatore,India.     

Effects of psoralen on replicon size and mean rate of DNA synthesis in partially synchronized cells of Pisum sativum L.

We have examined by fibre autoradiography the spacing of replicons in pea root meristems during synchronized entry into S phase from arrest at the G1/S boundary. Pretreatment with the DNA cross-linking agent, psoralen, produces a marked shortening of replicon spacing, suggesting that premature arrest of the replication fork results in the recruitment of additional initiation points within a given replicon family. This is discussed in relation to models for the control of DNA replication.     

Vitamin A effects on UMR 106 osteosarcoma cells are not mediated by specific cytosolic receptors.

Retinol and retinoic acid at 20 microM altered cell morphology and inhibited cell proliferation of UMR 106 osteosarcoma cells in culture. No specific cytosolic binding proteins for retinol could be detected.     

In vivo effects of pyrophosphate in Escherichia coli

Abstract Pyrophosphate (PP_i), a noncompetitive inhibitor of Rho poly(C)-dependent ATPase activity in vitro has been shown to relieve polarity in the lac operon. This indicates that PP_i could inhibit Rho activity in vivo too. An additional effect of PP_i on adenosine 3',5'-cyclic monophosphate (cAMP) synthesis during stationary phase of growth is also described.     

Circadian Rhythms in Competitive Sabre Fencers: Internal Desynchronization and Performance

During a 7-10 day span, circadian rhythms of sleep-wake, self-rated fatigue and mood, oral temperature, eye-hand skill and right and left hand grip strength were investigated in eight subjects: five males (21-28 years of age), members of the French sabre fencing team selected for the 1984 Olympic Games in Los Angeles, and three females (19-26 years of age) practicing fleuret (foil) fencing as a sports activity. On the average six measurements/day/variable/subject were performed. The single cosinor method showed that a circadian rhythm was detectable for only 26 out of the 56 time series (46.4%). Power spectrum analysis gave almost the same figure (19 out of 48: 39.5%) with regard to rhythms with τ=24hr indicating that with one exception (subject JFL) rhythms were internally desynchronized including differences τ between right and left hand grip strength rhythms for three subjects. Results suggest: (a) a physiologic synchronization of circadian rhythms may be a predictor of good performance; (b) however, internal desynchronization as shown previously may be a trivial phenomenon and thus does not imply in itself alterations of either health or performance; (c) chronobiologic methods should be recommended for a better understanding of changes in performance by those participating in competitive and other sports.     

Temporal Patterns of Reported Single-Vehicle Car and Truck Accidents in Texas, U.S.A. During 1980-1983

Reported single car and truck accidents due specifically to driver 'fatigue' for the 4-year span of 1980-1983 were analyzed for 24-hr, 7-d and 1-yr patterns. Three types of data were examined: (1) accidents in rural Texas, (2) accidents only in San Antonio, the fourth largest city in Texas and (3) accident ratio (number of accidents per time interval divided by traffic volume for the interval) in this city.

A statistically significant seasonal variation of relatively low amplitude with peaks in May and July was detected only for accidents in rural areas. High-amplitude periodicities of 7 d were evident in all the data sets, with a peak on the weekend. A very prominent 24-hr pattern was detected as well, with the acrophase determined by single cosinor around 0300. A slight secondary peak in accidents, around 1500, was evident in the data from the rural areas. This secondary peak represented a 12-hr rhythm in addition to the 24-hr. Overall, features of the temporal patterns in accidents over 24 hr and over the year in urban and rural areas differed only slightly. Accident ratios yielded results similar to accident frequencies. However, with regard to the rural data, the shape of the 24-hr temporal pattern varied with driver age; older drivers exhibited a flatter curve with a more pronounced afternoon rather than early morning (around 0300) peak.