Time course localization of immunoglobulin M monoclonal antibody and its fragments in leukemic tumor-bearing mice

Summary In vivo localization of a mouse monoclonal antibody (F₂-10.23 IgM) binding leukemic L 1210 cells was studied in DBA/2 mice bearing an L 1210 tumor. F(ab)₂ fragments were prepared and their specific binding to L 1210 cells was analyzed by flow cytofluorometry. Radiolocalization studies were performed by using ¹²⁵I- or ¹³¹I-labeled IgM monoclonal antibody or its F(ab')₂ fragments to ascertain their capacity to visualize the L 1210 tumor. F(ab)₂ fragments were cleared more rapidly than the whole IgM; the clearance was as fast in healthy as in tumor-bearing mice. The tumor-to-muscle ratio observed 24 h after injection of ¹²⁵I-radiolabeled F(ab)₂ fragments and ¹²⁵I-radiolabeled IgM was 10; the radioactivity level in the blood with F(ab)₂ fragments was lower than with IgM, and so -camera imaging was workable with F(ab)₂ fragments without background substraction. The tumor localization was studied over a period of 5 days by recording the distribution of the iodinated fragments in the tumor-bearing leg compared with that in the normal leg, and by computer analysis of the region of interest. F(ab)₂ fragments gave better results than intact IgM in tumor visualization. Nevertheless, the rapid clearance of this antibody or its F(ab)₂ fragments make them hardly suitable as carriers of toxic drugs. Abbreviations used are: MEM Minimum essential medium; SDS sodium dodecylsulfate; PAGE polyacrylamide gel electrophoresis     

Physiological characteristics of the tympanic organ in noctuoid moths

Summary The tympanic organ ofSpodoptera frugiperda, Mocis latipes, Erebus odorata (Noctuidae) andMaenas jussiae (Arctiidae) was stimulated with acoustic stimuli of 20 kHz, 45 ms and 5 s duration, and intensities ranging from 30 to 100 dB. The electric activity of the auditory receptors was recorded at the tympanic nerve with a stainless steel hook electrode. In all of these moth species there is an intensity range (ca. 20 dB) in which the response of each auditory receptor (A₁ and A₂ cells) to 45 ms pulses varies in a linear relation to the logarithm of stimulus intensity. For intensities higher than this value, depending on the species and the cell analysed, the spike discharge may continue to increase, may saturate or may diminish (Fig. 2). InE. odorata andM. latipes the A₁-cell response shows a decrease for stimulus intensities higher than 30 dB above the threshold. In the former species there is a statistically significant linear relation between the A₂-cell response and the decrease of the A₁-cell response, but this is not the case inM. latipes (Fig. 3). The similarity of the responses ofE. odorata to those described inEmpyreuma pugione (Coro and Pérez 1984) suggest that also in this noctuid species one may assume that the A₂ cell inhibits the A₁ receptor. In all of these moth species there is a maximum firing rate of the auditory cells at the beginning of the response to pure tones of 5 s and an exponential decrease of their discharge frequency with the course of time (Fig. 5). The analysed species differ in the adaptation rates of their auditory receptors. In all of these species the A₂ cell adapts more rapidly than the A₁ cell. In most of these species the stimulus intensity influences the adaptation rate of the auditory receptors (Fig. 7). These results are compared with data obtained by other authors, and it is concluded that there are more interspecific differences in the physiological characteristics of the auditory receptors in noctuoid species than those reported so far.Abbreviation AP action potential     

Lysosomal hydrolase activity in chorionic villi and embryonic cells in culture

Summary Fourteen lysosomal enzymes were compared in 20 cultured cell lines from chorionic biopsy and corresponding embryonic tissue after voluntary abortions. Enzymatic expression appears to be similar in cultured cells from both sources with some slightly higher levels for chorionic villi. We stress the importance of culturing chorionic villi especially in the case of enzymes (-L-iduronidase) or diseases (I cell disease) whose expression is unusual in fresh trophoblast tissue.     

Separation of isoenzymatic forms of human serum galactosyltransferase by chromatofocusing and isoelectric focusing. Application to cancerology

Chromatofocusing is used to separate the multiple isoenzyme forms of human serum galactosyltransferase. At least 11 major peaks of activity are observed in normal sera, which are eluted between pH 4.3 and 6.9; a fraction of activity is eluted above pH 7.0. The normal patterns are compared with those obtained with sera from cancer patients and with an ascitic fluid. Chromatofocusing appears as resolutive as agarose isoelectric focusing.     

Inactivation of soman by beta-cyclodextrin

Soman (pinacolyl methylphosphonofluoridate), a mixture of four stereoisomers, is inactivated appreciably in Tris buffer, pH 7.40, mu = 0.155 at 25 degrees C by beta-cyclodextrin (cycloheptaamylose, beta-CD). Under these conditions, the dissociation constant Kd of the 1:1 complex formed by beta-CD and soman and the rate constant k2 for the phosphonylation of beta-CD by soman are (0.53 +/- 0.05)mM and (5.9 +/- 0.6) X 10(-2) min-1 respectively. It results that the inactivation of soman by the mono-anion of beta-CD is about 2,600 times faster than the hydrolysis of soman by the hydroxide ion. The inactivation of both P(-) isomers of soman by beta-CD proceeds apparently at the same rate but both P(+) isomers react more slowly. Thus the interaction is stereospecific. The inactivation of soman by beta-CD appears to be as fast in human plasma in vitro as in Tris buffer.     

Repair of damage in double-stranded phi X174 (RF) DNA due to radiation-induced water radicals

Experiments in which the yields of radiation-induced OH and H radicals were varied, showed that both types of water radicals inactivate phi X174 RF DNA to about the same extent as measured by transfection of the (irradiated) DNA to E. coli wild-type spheroplasts. On the other hand, using spheroplasts prepared from E. coli strains, deficient in one of the proteins involved in excision DNA repair (uvrA- or uvrC-) or in post-replication repair (recA-), clear differences between damage originating from OH or H radical attack were found. Part of the radiation damage due to H radicals appeared to be repairable by an uvrA-gene-dependent repair mechanism, whereas this repair pathway does not play an important role in the case of OH radical damage. The reverse applies to uvrC-gene-dependent repair, which only affects OH radical damage (obtained under anoxic conditions), but has no influence on damage due to H radicals. Irradiation of double-stranded phi X174 (RF) DNA in the presence of oxygen however, yields damage--due to OH radicals only--which appeared not to be sensitive to either uvrC- or uvrA-gene-dependent repair. Furthermore, post-replication repair (recA) has only very little effect on the amount of inactivation by H or OH radicals, when irradiation is carried out under anoxic conditions. We did not find significant inactivation due to hydrated electrons, whether the biological activity was determined by use of wild-type spheroplasts or of strains deficient in excision or post-replication repair proteins.     

Expression of the gene encoding protein A in Staphylococcus aureus and coagulase-negative staphylococci

Two shuttle vectors containing the gene for protein A (spa) from Staphylococcus aureus have been constructed to study expression of the gene in various strains of S. aureus and in the coagulase-negative species Staphylococcus epidermidis, Staphylococcus capitis, and Staphylococcus xylosus. One plasmid, pSPA15, contains the complete structural gene for protein A, which binds to the cell wall in various Staphylococcus species. The other plasmid, pSPA16, codes for a truncated protein A lacking the C-terminal part called region X. The latter is exclusively extracellular in all Staphylococcus species tested, which confirms the importance of region X for cell wall binding. The expression of the plasmid-coded protein A in various strains of S. aureus is strongly correlated to the expression of the chromosomal spa gene. The coagulase-negative species expressing plasmid-encoded protein A produce 12 to 30% of the amount coded by the chromosomal spa gene in S. aureus strains Cowan I and A676.     

Protoplast forming ability in the genusBrevibacterium

Formation of protoplasts and their reversion were followed in 7 strains of brevibacteria. The formation of protoplasts and their reversion differed both between various species of brevibacteria and between various mutant strains of the same species.     

Malformation uropathies and multiple malformation syndromes

The authors, from their experience emphasize the associated malformations' frequency in major congenital urinary tract malformations (26,9%). It is essential to recognize in these multiple defects some certified syndromes - inherited or not. The most associations are still unknown, nevertheless the genetic counselling require an accurate diagnosis.     

Chromatographic behaviour of the molecular forms of guinea-pig skeletal muscle cytoplasmic malate dehydrogenase

The isolated molecular forms of guinea-pig skeletal muscle cytoplasmic malate dehydrogenase have a different chromatographic behaviour through affinity or hydrophobic interaction gels; in all cases the retention of the B form is more noticeable. Chromatography of a partly purified preparation through 5' AMP-Sepharose allows both molecular forms of malate dehydrogenase to be separated and obtained free from lactate dehydrogenase.