The usefulness of the micronucleus test using supravital staining of peripheral blood reticulocytes with acridine orange was evaluated in two laboratories after administering cyclophosphamide (CYP) as a model chemical by intraperitoneal injection (i.p.) to CD-1 mice. The frequencies of micronucleated peripheral reticulocytes (MNRETs) increased dose-dependently at each sampling time. There were no significant differences in the results obtained with this new method by the two laboratories. Although the induction of MNRETs was delayed by about 24 h compared to that of micronucleated polychromatic erythrocytes (MNPCEs) in the bone marrow, the frequencies of MNRETs and MNPCEs were almost identical at each optical sampling time, 24 h for MNPCEs and 48 h for MNRETs. Therefore, it is concluded that this method is a suitable alternative to that using femoral marrow cells.

Animals↗

PubMedDate not supplied

Frequency of anesthetic cardiac arrest and death in the operating room at a single general hospital over a 30-year period.

STUDY OBJECTIVES: To determine the anesthetic cardiac arrest (CA) and death rates in the operating room (OR) and to determine whether anesthetic CA and death are preventable. DESIGN: Prospective study. SETTING: OR suite of a general teaching hospital. PATIENTS: 85,708 patients undergoing surgery over a 30-year period. MEASUREMENTS AND MAIN RESULTS: Anesthetic CAs and deaths were identified, their causes and preventability were determined on the day of occurrence, and they were recorded over the years 1962 to 1992. Four anesthetic CAs (0.5 arrest per 10,000 anesthetics) and 1 death (0.1 death per 10,000 anesthetics) occurred during the 30-year study period. CONCLUSIONS: The results of 30 years' experience do not support the hypothesis that all anesthetic CAs and deaths are preventable. However, careful clinical management can reduce their frequency to a level lower than those reported in the referenced literature.

Adolescent↗

PubMedDate not supplied

Effects of total parenteral nutrition on endotoxin translocation and extent of the stress response in burned rats.

Postburn endotoxin translocation has been well documented. However, the relationship between the secretion of catabolic hormones, degree of endotoxin translocation, and intestinal atrophy has not been previously demonstrated. In this experiment, modulation of the secretion of catabolic hormones according to the route of nutrient administration was examined in burned animals. A total of 55 rats, with and without a burn injury, were orally or parenterally fed. Urinary excretion of epinephrine and norepinephrine (U-EN) of each rat was measured for 48 h after burn injury as an indicator of the stress response. Evaluations of intestinal atrophy and endotoxin contents in the liver and spleen were also done 48 h after burn injury. U-EN after burn injury in rats administered total parenteral nutrition (TPN) was higher than in those fed orally. Endotoxin translocation and intestinal atrophy after thermal injury were also augmented by TPN. A significant positive correlation between U-EN and endotoxin content of the liver, and a negative correlation between U-EN and weight of the intestine, were observed. TPN enhances the stress response after burn injury. An increase in endotoxin translocation and intestinal atrophy by TPN are closely related to enhancement of the stress response.

Animals↗

PubMedDate not supplied

A new sensitive and specific combination of CD81/CD56/CD45 monoclonal antibodies for detecting circulating neuroblastoma cells in peripheral blood using flow cytometry.

PURPOSE: Intensive chemoradiotherapy followed by peripheral blood stem cell transplantation has been introduced to treat children with advanced neuroblastoma (NBL). Detection of NBL cells in peripheral blood (PB) is important to prevent reinfusion of NBL cells. Several immunologic methods have been proposed for detecting NBL cells in hematologic samples. The development of a sensitive and specific combination of monoclonal antibodies (MoAbs) for detecting small numbers of NBL cells in PB using flow cytometry remains an important challenge. METHODS: Twenty-one clinical samples from NBL tissues or smears containing NBL cells were examined for reactivity against CD81, CD56, and CD9 using an immunocytochemical technique. The expressions of CD81, CD56, CD9, and antihuman disialoganglioside GD2 MoAb (GD2) in five NBL cell lines were assayed by flow cytometry. For the evaluation of sensitivity, five NBL cell lines were added to normal PB and the detection level of the combination of CD81/CD56/CD45 MoAbs was compared with that of CD9/CD56/CD45 MoAbs (reported previously). One hundred thirty-three normal PB samples were examined to determine the sensitivity and specificity of this method. RESULTS: All NBL cell lines showed strong positivity with CD81 and CD56 MoAb. However, CD9 MoAb was weakly positive against the five NBL cell lines. GD2 MoAb reacted strongly with four NBL cell lines, although almost the entire cell population of the SK-N-SH NBL line failed to bind the GD2 MoAb. In vitro experiments using NBL cell lines demonstrated that tumor cells added to normal PB cells could be detected by flow cytometry using CD81/CD56/CD45 MoAbs even at a concentration of 0.005%. Through comparative studies, the combination of CD81/CD56/CD45 MoAbs was found to be more sensitive and specific than that of CD9/CD56/CD45 MoAbs for detecting small numbers of NBL cells using the above cell lines. CONCLUSIONS: Triple-color flow cytometric analysis using CD81/CD56/CD45 MoAbs is useful for detecting NBL cells in PB. Further studies testing this approach using samples of PB with NBL contamination are needed to test this approach in patients.

Adolescent↗

PubMedDate not supplied

Unresponsiveness to surface antigen 1 modifies cytokine profiles in acute Toxoplasma gondii infection.

Resistance to Toxoplasma gondii involves the development of a highly polarized Th1-type cytokine expression. SAG1 transgenic mice are highly susceptible to T. gondii infection due to their non-reactivity to SAG1 of the protozoan parasite. Here we describe cytokine profiles during the acute phase of T. gondii infection, which are associated with the susceptibility of SAG1 transgenic mice. SAG1 transgenic mice showed a 4.5-fold increase in susceptibility upon inoculation with a sublethal dose of the Beverley strain of T. gondii compared to their wild-type counterparts (mortality: 81 vs. 18%, respectively). When analysis of the most important cytokines involved in the mediation of resistance to infection was carried out, SAG1 transgenic mice exhibited low production levels of IL-12, IFN-gamma and TNF-alpha in sera during the acute phase of T. gondii infection. Antibody and T cells specific for SAG1 were not mounted upon SAG1 stimulation in SAG1 transgenic mice. Moreover, in vitro studies indicated that in SAG1 transgenic mice IFN-gamma and IL-12 production was lower than in their wild-type counterparts, although levels of TNF-alpha increased in SAG1 transgenic mice on day 9 after infection. Low IgG2a levels were detected in SAG1 transgenic mouse sera. Unresponsiveness to SAG1 of T. gondii renders SAG1 transgenic mice unable to develop a strong Th1-based protection against T. gondii infection. These results provide evidence that SAG1 is a pivotal antigen involved in the induction of immune responses towards the development of Th1-protective immunity during T. gondii infection.

Animals↗

PubMedDate not supplied

Quantitative analysis of kanamycin ototoxicosis.

The morphological changes after kanamycin intoxication of the inner ear, including both the cochlea and the vestibule, were quantitatively analysed by the surface preparation technique after succinic dehydrogenase staining. 75 guinea pigs were used. The outer hair cells in the basal coil and the inner hair cells in the upper coils of the cochlea were the most severely damaged, but many unusual modes of damage were also revealed. For example, the initial hair cell damage in the cochlea appeared in the upper hair cells. The clearly observed vestibular damage contradicts the general belief that kanamycin is not so toxic to the vestibular hair cells. The utricular macula and the lateral crista were most severely damaged. The delayed ototoxicity of kanamycin was observed for the first time in the vestibular hair cells.

Animals↗

PubMedDate not supplied

Tissue levels of kanamycin in correlation with oto- and nephrotoxicity.

The time courses of kanamycin (KM) levels in the perilymph, kidney, liver, brain and blood were observed by a bioassay method in normal guinea pigs and in animals pretreated with KM for 10 successive days. In the kidney and perilymph of the normal guinea pigs, the injected KM became highly concentrated and was eliminated only after a protracted delay. This tendency was much more intensified in kidney and perilymph of the pretreated animals. By contrast, the KM in blood and brain was eliminated rapidly both in the normal and in the pretreated animals. It is concluded that the high accumulation and slow elimination of KM by the inner ear and kidney and, moreover, the enhancement of these phenomena by successive pretreatment are one of the important factors in the origin of oto- and nephrotoxicity.

Animals↗