Immunocytochemical study of the action of cis-dichlorodiamino platinum on the human metaphase chromosome.

The reaction of cis-dichlorodiamino platinum (cis-Pt) with the human metaphase chromosome was studied using both Giemsa staining and the action of anti-cis-Pt antibody. Two kinds of experiment were performed: (1) cis-Pt was placed in contact with cells in culture (in vivo tests), (2) cis-Pt was placed in contact with chromosomes fixed on slides (in vitro tests). In vivo, the presence of cis-Pt during phase G1 of the cell cycle is important for the reaction; in vitro, competition between cis-Pt and Giemsa for fixation on chromosomal DNA was shown to occur.

Reactivity of anti nucleoside antibodies with metaphase chromosomes.

It is shown that the binding of anti-nucleoside antibodies to fixed human metaphase chromosomes can be revealed using the immunoperoxidase procedure while under the same conditions no antibody binding is revealed using the immunofluorescence procedure.

Measurement of glutamine synthetase activity in rat muscle by a colorimetric assay.

Glutamine synthetase catalyses the formation of L-Gln from L-Glu and NH4+. This enzyme also exerts a glutamyl-transferase activity that produces gamma-glutamyl-hydroxamate from Gln and hydroxylamine. This gamma-glutamyl-transfer reaction can be used to determine glutamine synthetase activity by colorimetric assay. This method has never been applied to rat muscle. The aim of this work was to study and optimize the glutamine synthetase assay conditions in rat muscle. Enzyme activity was linear with time of incubation (30 min at 37 degrees C) and linear with enzyme concentration in the incubation medium. The method was specific. In addition, this assay correlated well with a radiometric assay (y = 0.76x + 340, where x and y are the glutamine synthetase activities measured by radiometry and colorimetry respectively; r = 0.94; P = 0.05). Finally, no glutamine synthetase activity was found in muscles of rats treated with methionine sulfoximine, an inhibitor of glutamine synthetase, and activity dramatically rose in muscles from rats treated with dexamethasone, an activator of glutamine synthetase (in extensor digitorum longus: 2717 +/- 54 nmol/min/g protein in dexamethasone-treated rats versus 1228 +/- 114 nmol/min/g protein in control rats, P < 0.0001). In conclusion, the method presented here is accurate and reliable for measurement of glutamine synthetase activity in muscles.

Supplementation of oral nutrition with pancreatic enzymes improves the nutritional status of aged endotoxemic rats.

Malnutrition is a common problem in elderly people. The association of malnutrition and physical illness or injury leads to both localized and general complications. In particular, impairment of the adaptive response of pancreatic function to undernutrition and refeeding may adversely affect nutritional status and elicit morbidity and mortality. Aged rats (24 mo old) were treated with lipopolysaccharide (LPS) from E. Coli (3 mg/kg body weight). Six days later, survivors were randomized to receive, for 7 days, an oral chow diet enriched with either a pancreatic extract (PE) (2.4 mg/day) or an isonitrogenous supply of casein (CAS). Endotoxemia induced a catabolic state, with a body weight loss of 7.6 +/- 1.1% on day two after LPS treatment. Mean food intake from day 6 to day 13 was similar in LPS-PE and LPS-CAS groups (19.0 +/- 5.6 versus 19.7 +/- 6.9 g). The metabolic response varied according to the type of muscle studied. In fast (white) muscle, the protein content and the glutamine pool remained markedly depleted in endotoxemic rats receiving casein supplementation. In contrast, enrichment of nutrition with PE significantly limited the LPS-induced muscle wasting and increased the muscle glutamine content. As in previous observations, no significant change occurred in slow (red) muscle. These results could indicate that PE supplementation counteracts pancreatic deficiency caused by aging and worsened by stress and this, in turn, could improve the efficiency of nutrition, to support the hypermetabolism of aged injured rats.

Comparison between two chromogenic substrates for revealing an immunoperoxidase reaction of human metaphase chromosomes.

To study the binding of an antiadenosine serum to human chromosome DNA, two types of chromogenic reagents were compared. The procedure is as follows: lymphocytes with metaphase chromosomes were spread on slides; some slides were irradiated by UV; all slides were then incubated with antiadenosine rabbit serum and then with antirabbit sheep serum labelled with peroxidase; the latter was revealed in the classical manner by 3,3'-diaminobenzidine (DAB), or, alternatively, by p-phenylenediamine plus pyrocatechol (PPD-PC). The present study shows that the results obtained with PPD-PC were equivalent, if not superior, to those obtained with DAB. In the case of weaker reactions, results with PPD-PC were superior. Furthermore, this reagent has the major advantage of being noncarcinogenic.

Effects of orchiectomy on EDL muscle ultrastructure and energy-supplying enzymes in growing male guinea pigs.

To investigate the influence of androgens on muscle maturation during growth, guinea pigs were orchiectomized (CX) or sham operated (SO) at the end of weaning. Extensor digitorum longus (EDL) muscles were removed at weeks 4, 8, and 12 of postnatal development. According to their contractile and metabolic characteristics, four fiber types were distinguished, called I, IIa, IIbox, and IIbglyc. Muscle maturation consisted of a concomitant decrease in percentages of type IIa and IIbglyc fibers and increase in the percentage of IIbox fibers in both groups. At week 12, fiber distributions were not different between the two groups. Citrate synthase activity fell and phosphofructokinase and lactate dehydrogenase (LDH) activities rose from week 4 to week 12 and were the same in CX and SO. Muscular LDH subunits increased in SO and decreased in CX during this period. In conclusion, fiber type distribution and enzyme activities at puberty were not androgen dependent in guinea pig EDL muscle. Conversely, these hormones acted on LDH isozyme distribution through the enhancement of the most glycolytic LDH fractions.

[Immunocytochemical labeling of human chromosomes by antibodies from human autoimmune serum]. / Marquage immunocytochimique de chromosomes humains après action d'anticorps de sérum autoimmun d'origine humaine.

Mitotic human chromosomes are successively irradiated by ultraviolet, incubated with serum from a patient showing systemic lupus erythematosus (S.L.E.), and antihuman Sheep serum conjugated with peroxidase. After revelation by diaminobenzidine (DAB) banding patterns are pointed out. The chromosome labelling is discussed in comparison with results obtained in other immunocytochemical experiments.