Identification of oral species of the genus Veillonella by polymerase chain reaction.

INTRODUCTION: Members of the genus Veillonella cannot be reliably distinguished by their biochemical characteristics and phenotypic features. Moreover, DNA-DNA hybridization and sequence analyses of the 16S ribosomal RNA gene including random fragment length polymorphism analysis, are complex and time-consuming procedures that are not well-suited to identifying oral species of Veillonella: Veillonella atypica, Veillonella denticariosi, Veillonella dispar, Veillonella parvula, and Veillonella rogosae. METHODS: In this study, five forward primers and a reverse primer were designed for polymerase chain reaction (PCR) according to the partial sequences of the rpoB genes of these oral Veillonella species. RESULTS: The forward primers were species-specific for these five Veillonella species, and could produce specific amplicons when used together with reverse primer and individual DNA templates of these species in PCR. These primer pairs were also found to discriminate between the respective species, and the Veillonella strains isolated from human oral cavities were successfully assigned to one of the five oral species of the genus Veillonella based on their specific products by PCR. CONCLUSION: A simple two-step PCR procedure using the five sets of primer pairs developed in the present study is a rapid and reliable method for the identification of the recognized oral Veillonella species.

Prognostic predictive values of serum cytochrome c, cytokines, and other laboratory measurements in acute encephalopathy with multiple organ failure.

AIMS: To evaluate the prognostic predictive values of cytochrome c, cytokines, and other laboratory measurements in serum collected during neurological onset in acute encephalopathy with multiple organ failure. METHODS: In addition to general laboratory examinations, the concentrations of cytochrome c (apoptosis marker) and cytokines (inflammatory markers) were measured in serum samples collected at the initial phase in 29 patients with acute encephalopathy. The obtained values were evaluated as predictors for the development of severe encephalopathy. RESULTS: Cytochrome c, tumour necrosis factor alpha (TNF-alpha), interleukin 6 (IL-6), soluble TNF-receptor 1 (sTNF-R1), and aspartate aminotransferase (AST) concentrations at the initial phase were high and correlated well with patient outcome. High concentrations of serum cytochrome c (>45 ng/ml), sTNF-R1 (>2000 pg/ml), AST (>58 IU/dl), IL-6 (>60 pg/ml), and TNF-alpha (>15 pg/ml) predicted an unfavourable prognosis (sequelae and death) at 93%, 79%, 82%, 77%, and 60%, respectively. The specificity of those markers was 100%, 89%, 83%, 100%, and 100%, respectively. CONCLUSIONS: Serum cytochrome c is the most sensitive and specific predictor for the development of severe encephalopathy at the initial phase. Results suggest that this marker might be used to guide decisions regarding the start of the initial treatment and further intensive care.

Effects of illumination and handling upon rat open field activity.

Eighty male rats (Crj:CD, 4 weeks old), were randomly assigned to four groups in a 2 x 2 (brightness x handling) factorial design. Ambulation and three types of defecations [pre-open-field defecation (PRD), post-open-field defecation (POD), and testing defecation (TD)] were examined for 3 consecutive days, for 3 min per day. In this study, the effects of handling treatment were illumination dependent. The changes in TD and POD were opposite in relation to the trial day, whereas the changes during the trial, the sum of the TD and POD, were invariable. Considering the invariable changes of the sum of TD and POD, time per trial may be an important factor to measure emotional activity. The TD-POD inversion may reflect the strength of the stress response against a novel placement in open field, because of the change of the time for evoking defecation behavior.

A method for detecting visceral malformations in gelatin-embedded rat fetuses using an automatic slicing apparatus.

A method for observing whole rat fetal viscera embedded in gelatin using an automatic slicing apparatus is described. Fetuses were immersed in Bouin's solution. Part of the thoracic and abdominal skin of each fetus was removed, and fetuses were immersed consecutively in sodium bicarbonate 30% in 70% ethanol, gelatin 15% in water, gelatin 30% in water, then embedded in fresh 30% gelatin. The gelatin blocks containing the fetuses were immersed in 10% formalin. After fixation, the block was sliced into 200 microns serial transverse sections using a rotor-slicer at a rotation speed of 120 rpm and a cutting speed of 25 mm/sec. Complete slicing of a single fetus required about 20 min. The advantages of the method presented here include: complete fetal serial sections are produced, thin and uniform sections are obtained easily, viscera can be identified easily, and observation can be carried out at any time after slicing. The method presented can be used to detect whole fetal visceral malformations in developmental toxicity tests.

Influence of electrocardiographic findings on admission decisions in patients with acute chest pain.

This study prospectively evaluated the influence of current electrocardiograms obtained at the time of emergency department presentation, as well as that of previous comparison electrocardiograms, on decision-making regarding coronary care unit admission of patients presenting with a chief complaint of chest pain or chest pain equivalent. Emergency department physicians were asked to commit themselves to recommending either coronary care unit admission or some other disposition, both before and after evaluating current comparison electrocardiographic findings. They were also asked, prior to reviewing these results, whether they thought information gained from the electrocardiograms would have any affect on their decision. Despite wide expectation that electrocardiographic findings would in fact affect decision-making, neither current nor comparison electrocardiograms virtually ever altered the ultimate decision of whether or not to admit. Faculty and house officers performed similarly in all regards, except insofar as attending physicians were less likely to expect electrocardiographic findings to help them in patients who were ultimately discharged. Emergency department nurses, who were asked whether they believed these patients needed admission to a coronary care unit on the basis of only a brief initial triage history, performed very similarly to the physicians. Thus, electrocardiographic findings are rarely if ever helpful in determining the need for admission to a coronary care unit in patients presenting to the emergency department with chest pain, and seem to have particularly little value in patients in whom myocardial infarction is considered clinically unlikely. Although physicians at all levels of training often feel a need to rely on electrocardiograms in patients they ultimately admit, greater experience allows more senior physicians to be comfortable in correctly discharging patients with no clinical evidence of disease without obtaining an electrocardiogram. Routine ordering of electrocardiograms in patients with chest pain in whom likelihood of significant acute ischemic pain is clinically low should be reconsidered.