Association between the neutrophil myeloperoxidase index and subsets of bacterial infections.

The mean myeloperoxidase index (MPXI) is calculated during the routine complete blood count performed using the autoanalyzer ADVIA120/2120. The pattern of changes in the neutrophil myeloperoxidase levels in patients with specific infectious diseases was analyzed by assessing the MPXI levels. In patients with bacterial sepsis, identified by positive blood-culture tests, with (n = 29) and without (n = 51) systemic inflammatory response syndrome, the mean MPXI significantly reduced to -3.18 and -2.06, respectively. In contrast, among patients with nontuberculous nonseptic bacterial infections (n = 40), the mean MPXI significantly elevated to 5.51, while tuberculosis patients (n = 37) and patients with viral infection (n = 60) showed an unchanged MPXI (mean values, -0.46 and -1.06, respectively). Among the parameters of inflammation, only the C-reactive protein values showed a weak correlation with the MPXI levels. [Conclusion] These results indicate that MPXI is correlated with some specific infectious states, i.e. MPXI is low in bacterial sepsis and high in nontuberculous nonseptic bacterial infections. MPXI appears to be an independent and useful biomarker for the diagnosis and follow-up of infectious diseases, especially when the MPXI values are obtained at regular intervals during the disease courses of the patients.

Relations of the c-myc gene and chromosome 8 in non-small cell lung cancer: analysis by fluorescence in situ hybridization.

BACKGROUND: Amplification of the c-myc gene has been reported in non-small cell lung cancer (NSCLC). We investigated the c-myc gene amplification and the numerical aberration of chromosome 8 by dual color fluorescence in situ hybridization (FISH) to evaluate the relation between possible genetic abnormalities, pathological factors and prognosis. METHODS: Tumor tissue samples were obtained from 31 patients with NSCLC who underwent lobectomy with mediastinal lymph node dissection. Samples were analyzed by FISH using 8 alpha satellite DNA probe and c-myc gene cosmid probe. The relation between genetic abnormalities, pathological factors (T factor, tumor size, and N factor), and prognostic factors was evaluated by univariate and multivariate analysis, and by the Kaplan-Meier method and log-rank analysis. RESULTS: Chromosome 8 aberrations were T1 (n=3), 44.0%; T2 (n=18), 35.7%; T3 (n=7), 40.0%; T4 (n=3), 39.7% (p=NS). The c-myc gene amplifications were T1, 54.3%; T2, 51.1%; T3, 51.0%; T4, 66.3% (p=NS). There was no difference between patients whose tumor was more than 5 cm (n=16), and 5 cm or less (n=15) in the rate of chromosome 8 aberration (39.3%: 36.3%), or the rate of the c-myc gene amplification (52.1%: 53.7%). N factors for chromosome 8 aberrations were N0 (n=18), 35.9%; and N2 (n=11), 44.9% (p=NS). In the c-myc gene amplification, there was a significant difference between N0 and N2 (48.6%, 61.3%, p=0.040). In univariate and multivariate analysis, chromosome 8 aberrations correlated with a poor prognosis (p=0.037 and p=0.041). The 5-year survival rate was 15.4% in patients whose rate of chromosome 8 aberrations was 40% or more (n=13), which was significantly less than that in patients with an aberration rate of less than 40% (n=19, 57.9%, p=0.014). CONCLUSION: The c-myc gene amplification correlates with lymph node metastasis. Although there was no significant link between the amplification of the c-myc gene and clinical outcome, the numerical chromosome 8 aberrations was considered to be a factor for survival.

Reliability of chromosome aberration by one-color fluorescence in situ hybridization.

The fluorescence in situ hybridization method allows the observation of chromosomal aberrations under a microscope at the cellular level. However, the extent to which the FISH method reflects actual chromosomal aberrations is unknown. To estimate the accuracy of detecting aberrations by FISH, we performed dual color-FISH with two different DNA probes for the principal target DNA and assessed their concordance. The two DNA probes used were a whole chromosome painting probe and an alpha satellite probe. A high concordance rate of 82%-98% was found between the probes, indicating that the accuracy of determining chromosomal aberrations by FISH is high.

Low level c-myc gene amplification in gastric cancer detected by dual color fluorescence in situ hybridization analysis.

BACKGROUND AND OBJECTIVES: By using the dual color fluorescence in situ hybridization analysis, the amplification of c-myc gene can be detected in the tumor tissue samples obtained from patients with gastric cancer, and the relationship between the molecular cytogenetic change and the clinical stage or histological type may be clarified. METHOD: The tumor tissue samples were obtained from 21 patients with gastric cancer. Simultaneous detection of signals from the chromosome 8 centromere and c-myc gene in each cell after hybridization with appropriate probes was carried out on 50-200 tumor cells in each case. RESULTS: Chromosome 8 polysomy was found in 10 patients. The average centromere 8 copy number was significantly higher in differentiated (2.7) than in undifferentiated (2.3) types of gastric cancer. However, there was no significant difference in occurrence of polysomy 8 between early and advanced cancer. The relative gain (1.1-1.9) of c-myc copy number was found in all 21 cases. There was no significant difference in the fraction of cells with c-myc gene amplification between early (pT1) and advanced (pT2-4) carcinomas. CONCLUSION: We conclude that the present dual color fluorescence in situ hybridization of gastric cancer may be useful in the evaluation of low level c-myc gene amplification, which is difficult to detect by Southern blotting, and may be applicable to the diagnosis of early gastric cancer.

Chromosome 17 copy numbers and incidence of p 53 gene deletion in gastric cancer cells. Dual color fluorescence in situ hybridization analysis.

Tumor tissue samples were obtained from each of ten patients with stomach cancer who had undergone surgery. The simultaneous detection of signals from 17 alpha pericentromeric and 17 p 13.1 loci after hybridization with the appropriate probes was carried out on 100-200 tumor cells in each case. The copy number category (i.e., monosomy, disomy, trisomy, and so on) was defined as described by Waldman et al., and the deletion of a gene of interest was defined as described by Matsumura et al. The copy number category was monosomic in seven and disomic in three out of ten cases. The frequency of the copy number 2 (two centromeric signals per nucleus) was lower in tumor cells than in control cells that comprise normal gastric epithelium cells and cells from a normal fibroblast cell line. The copy numbers 3, 4, 5, and above (polysomy) were more frequent in tumor cells, whereas no normal cells having three or more centromeric signals per nucleus was present in the control. Incidence of p 53 gene deletion in ten patients ranged from 55.4% to 90.0%, and from 1.5% to 30.7% if nuclei having three or more copy numbers were taken into consideration. The mean incidence of the p 53 deletion was significantly higher in the differentiated type than in the undifferentiated type of carcinoma (p < 0.02). Various types in the combination pattern of 17 alpha centromere/17 p 13.1 signals were observed, and the most frequent pattern in the ten cases was a combination of 1/1. The deletion of chromosome 17 p may play an important role in carcinogenesis of the stomach.

Primary aortoduodenal fistula caused by duodenal tuberculosis without an abdominal aortic aneurysm: report of a case.

A 66-year-old man died of massive gastrointestinal hemorrhage caused by a fistula between the third portion of the duodenum and the abdominal aorta. An autopsy revealed that duodenal tuberculosis had resulted in the development of a fistula into the aorta with no pathological changes, and no active pulmonary tuberculosis was found. Duodenal tuberculosis and primary aortoduodenal fistula (ADF) without an aneurysm are both extremely rare. Thus, we report herein a unique case of primary aortoduodenal fistula without an abdominal aortic aneurysm, but associated with duodenal tuberculosis, and review the current literature.

Applications of fluorescence in situ hybridization in biological dosimetry and detection of disease-specific chromosome aberrations.

Dual color FISH with whole chromosome and pan-centromere probes facilitates rapid detection of stable structural aberrations such as translocations. This approach should allow analysis of translocations for assessment of genetic damage at long times after exposure or as a result of chronic exposure during a long period of time. Multi-color FISH with locus specific probes allows assessment of the frequency of cells carrying specific aberrations known to be associated with tumorigenesis, analysis of the series of genetic changes that occur during tumor evolution and correlation between genotype and phenotype. The power of FISH for analysis of random and tumor related events will increase steadily as informative probes are developed during the course of the International Human Genome Project.

[Effect of adriamycin on cell cycle by bromodeoxyuridine-labeling method].

We studied the effect of adriamycin on DNA synthesizing cells from the view point of cell cycle. DNA synthesizing cells were marked by bromodeoxyuridine, which is a known marker for such cells, before administration of adriamycin. Then, the level of this marker was sequentially measured by flow cytometry. At the low concentration (0.01 micrograms/ml), adriamycin caused delay of the shifting time to the S phase 16 hours after administration. The marked cells were accumulated at the G2M phase at a moderate concentration (0.1 micrograms/ml), and blocked at the S phase at the higher concentration (1.0 micrograms/ml).

Rapid human chromosome aberration analysis using fluorescence in situ hybridization.

We have used in situ hybridization of repeat-sequence DNA probes, specific to the paracentromric locus 1q12 and the telomeric locus 1p36, to fluorescently stain regions that flank human chromosome 1p. This procedure was used for fast detection of structural aberrations involving human chromosome 1p in two separate experiments. In one, human lymphocytes were irradiated with 0, 0.8, 1.6, 2.4 and 3.2 Gy of 137Cs gamma-rays. In the other, human lymphocytes were irradiated with 0, 0.09, 0.18, 2.0, 3.1 and 4.1 Gy of 60Co gamma-rays. The frequencies (per cell) of translocations and dicentrics with one breakpoint in 1p and one elsewhere in the genome were determined for cells irradiated at each dose point. These frequencies both increased with dose, D, in a linear-quadratic manner. The delta, alpha, and beta coefficients resulting from a fit of the equation f(D)=delta + alphaD + betaD2 to the translocation frequency dose-response data were 0.0025, 0.0027 and 0.0037 for 137Cs gamma-rays, and 0.0010, 0.0041, and 0.0057 for 60Co gamma-rays. The delta, alpha, and beta coefficients resulting from a fit to the dicentric frequency dose-response data were 0.0005, 0.0010 and 0.0028 for 137Cs gamma-rays and 0.0001, 0.0002 and 0.0035, for 60Co gamma-rays. Approximately 32,000 metaphase spreads were scored in this study. The average analysis rate was over two metaphase spreads per minute. However, an experienced analyst was able to find and score one metaphase spread every 10s. The importance of this new cytogenetic analysis technique for biological dosimetry and in vivo risk assessment is discussed.

[Estimation of cell viability by FCM using rhodamin 123 and propidum iodide--its application in evaluating the effect of anticancer drugs to tumor cells].

Dual parameter analysis of FCM using rhodamine 123 (R 123) and propidium iodide (PI) has made it easy to estimate cell viability. PI stains only dead cells, whereas R 123 accumulates in the mitochondria of living cells which remain unstained by PI. Therefore the combination of R 123 and PI provides a more accurate estimation of cell viability than the conventional methods of dye exclusion. Cell viability has been estimated by this method in HeLa cells treated with adriamycin in vitro. The usefulness of this analysis is suggested for the evaluation of the effect of anticancer drugs on tumor cells.

[Limits of detection of nuclear DNA aneuploidy by FCM].

The limits of FCM detection of nuclear DNA aneuploidy were determined by simulation of the relationship between the coefficient of variation (CV) and DNA difference of two populations using a microcomputer. Under ideal conditions, the minimal DNA difference necessary for a bimodal peak is almost twice the CV. When the DNA difference is below the value, a single peak with a large CV and/or a shoulder is formed. Cases with a larger CV than expected in a single population should be practically treated as a population mixed with the near-diploid line.

A physiological study of acoustic blinking reflex in the cat.

Blinking reflex was elicited by acoustic stimulus to investigate the reflex pathway in the encéphalo isolé cat. Reflex responses were recorded using electromyography (EMG) of the orbicularis oculi. The investigation was as follows: After stimulation by a loud click sound, the EMGs of the orbicularis oculi were elicited bilaterally. Latencies were 17-20 msec on the ipsilateral side, and 19-21 msec on the contralateral side. By electrical stimulation of the peripheral nerve in the cochlea and the acoustic nerve EMGs were also obtained. The latencies of the electrical stimulation responses correlated well with the latencies in the EMGs obtained through sound stimulation. The evoked potentials after auditory and electrical stimulation were obtained in several nuclei in the brainstem. The conduction time between two nuclei was calculated by measuring the latency of discharges in each nucleus. The alteration of the evoked discharge was observed through surgical transection of the brainstem. After each successive surgery the EMG showed differences in responses. Thus, the location of the neural connections of the pathways were differentiated in the brainstem. From this, we concluded that the reflex arc has the following pathway: The acoustic input to the ventral cochlear nucleus (VCN) is relayed to the acoustic route of the superior olivary complex and to the lateral lemniscus. Then, the impulse reaches the VIIth nucleus via the pontine reticular formation in the ipsilateral side. On the other hand, the input to the VCN crosses over the trapezoid body to the superior olivary complex. Then, the impulse uses the same pathway as described for the contralateral side. Finally, the pathway of the acoustic blinking reflex is located more caudally than that of the tactual blinking reflex. The comparison of the recordings of both reflexes is useful clinically, as a diagnostic method to study the function of the brainstem.