Indoxyl-UDPG-glucosyltransferase from Baphicacanthus cusia.

The enzyme catalyzing the transfer of glucose from uridine diphosphate glucose to indoxyl yielding the indoxyl glucoside indican was isolated from Baphicacanthus cusia Bremek (Acanthaceae). The indoxyl-uridine diphosphate glucose (UDPG)-glucosyltransferase was purified to homogeneity in six chromatographic steps. The decisive step for the recovery of a homogeneous enzyme was the application of immobilized metal affinity chromatography yielding an 863-fold purified enzyme. From a total of 60 substances tested, in addition to the natural substrate 3-OH-indole (indoxyl), only 4-OH-, 5-OH-, 6-OH-, and 7-OH-indole were accepted as substrates by the glucosyltransferase. However, the latter substrates were metabolized to varying extent. The optimum pH of the enzyme was 8.5, the optimum temperature was 30 degrees C and the isoelectric point was pH 6.5. The M(r) of the enzyme was determined to be 60 +/- 2 x 10(3). Indoxyl as substrate yielded a K(m) of 1.2 mM, while a K(m) of 1.7 mM was found for UDPG.

Enterococcus faecalis gene transfer under natural conditions in municipal sewage water treatment plants.

The ability of Enterococcus faecalis to transfer various genetic elements under natural conditions was tested in two municipal sewage water treatment plants. Experiments in activated sludge basins of the plants were performed in a microcosm which allowed us to work under sterile conditions; experiments in anoxic sludge digestors were performed in dialysis bags. We used the following naturally occurring genetic elements: pAD1 and pIP1017 (two so-called sex pheromone plasmids with restricted host ranges, which are transferred at high rates under laboratory conditions); pIP501 (a resistance plasmid possessing a broad host range for gram-positive bacteria, which is transferred at low rates under laboratory conditions); and Tn916 (a conjugative transposon which is transferred under laboratory conditions at low rates to gram-positive bacteria and at very low rates to gram-negative bacteria). The transfer rate between different strains of E. faecalis under natural conditions was, compared to that under laboratory conditions, at least 10(5)-fold lower for the sex pheromone plasmids, at least 100-fold lower for pIP501, and at least 10-fold lower for Tn916. In no case was transfer from E. faecalis to another bacterial species detected. By determining the dependence of transfer rates for pIP1017 on bacterial concentration and extrapolating to actual concentrations in the sewage water treatment plant, we calculated that the maximum number of transfer events for the sex pheromone plasmids between different strains of E. faecalis in the municipal sewage water treatment plant of the city of Regensburg ranged from 10(5) to 10(8) events per 4 h, indicating that gene transfer should take place under natural conditions.

Mega code evaluation: the development of graded criteria.

We developed an objective evaluation tool for grading students' performances in the advanced cardiac life support mega code. Specific functions for each student role are delineated. Each function is linked to a point system yielding an actual grade for each role, and pass/fail criteria are based on this grade. Use of the tool can eliminate instructor subjectivity and provide a permanent record of the student's performance.

Surgical treatment of ulcerative colitis in children.

Our finding in 100 patients indicate that elective surgical treatment of ulcerative colitis in children has a low mortality rate in contrast to emergency surgery (2.3% versus 23%). Morbidity associated with these procedures is moderate and appears to be decreasing (13%), as shown by the comparison between the experience in the 1960s and that in the 1970s. Results with the newer surgical procedures have been encouraging; 60% of patients with Brooke ileostomy have been very satisfied as compared with 90% of patients with Kock pouch and of those with rectal mucosectomy with ileoanal anastomosis. Ileorectostomy was not a satisfactory procedure in this series. Additionally, after surgical treatment, the median height percentile increased from the thirty-ninth to fifty-seventh percentile. Finally, and important, 96% of the patients in this series reported that their general health at follow-up was good to excellent and that they had minimal limitation in their activities.

Myocardial infarction following coronary artery bypass: factors influencing its occurrence.

The authors have undertaken a prospective study on postoperative myocardial infarciton (PMI) following coronary artery bypass (CAB) grafting. PMI was diagnosed from electrocardiographic findings by the criteria of the American Heart Association. From Aug. 1, 1975 to Feb. 27, 1976, 198 patients (177 men, 21 women) underwent CAB. Their ages ranged from 31 to 71 years (mean, 49 yr). Of these patients, 18 (9%) sustained a PMI. A number of factors were analyzed to determine their influence on the occurrence rate of PMI. Preoperative factors wuch as the New York Heart Association classification, measurement of left ventricular function and evaluation of the severity of coronary artery disease were not helpful in predicting a predisposition to PMI. Endarterectomy was the only operative intervention associated with a higher inicidence of PMI (18%). Other factors such as the interval of cardiac anoxia and cardiopulmonary bypass and the number of vessels grafted did not seem to be associated with PMI. Low blood flow (less than 40 ml/min) through the graft could be correlated with the area of infarction in only 5 of 18 patients. Among the operated patients, postoperative ventricular arrhythmia was much more common in patients with PMI. The hospital mortality in this group was 11% (two patients).

A quantitative analysis of oxygen release due to changes in P50 and venous PO2.

It is almost impossible to estimate the amount of oxygen released from hemoglobin on the basis of the P50 values without calculation, because of the shape of the oxygen dissociation curve. For this reason, we prepared two tables : one indicating the saturation of hemoglobin with oxygen in the venous blood as influenced by venous Po2 and P50 (Table I) and the other, the oxygen release from the arterial blood as influenced by venous Po2 and P50 (Table II). Both tables are based o- the Hill's equation and normal entry data. Values of P50 vary from 20 to 45 torr, while the range for venous Po2 is 5-50 torr. The discussion on possible use of the tables follows.