Determination of plasmid copy number in yeast transformants by means of agarose plugs

The determination of plasmid copy number in Saccharomyces cerevisiaetransformants containing circular or linear plasmids is currently performed with total yeast DNA extracts obtained from cultures grown under selection. The determination is based essentially on quantitative Southern hybridization of an appropirate probe to a sequence present both on plasmid and chromosomal DNA in digested or undigested samples run out on conventional agarose gels. The DNA extraction procedure calls for treatment of cell lysates with organic solvents that could entail systemic losses of eithr plasmid or chromosomal DNA thus producing artifactual results. We propose here a method based on the assumption that quantitative analysis of plasmid and chromosomal DNA extracted from yeast cells embedded in agarose plugs will furnish more reliable results. With this procedure the cells are lysed in situ, thus avoiding possible losses of material, and the chromosomes and plasmid DNAs, trapped within the agarose matrix, can be separated by pulse field electrophoresis

Apuleius

Die Schrift des Rhetors und platonischen Philosophen Apuleius von Madaura (Nordafrika, 2. Jh. n.Chr.) ist ein öffentlich gehaltener Lehrvortrag, in dem der Autor der in der Antike vielverhandelten Frage nachgeht, was das sog. Daimonion sei, auf das Sokrates sich in den Platonischen Dialogen und in den Schriften Xenophons so häufig beruft. Um diese Frage umfassend zu beantworten, entwirft Apuleius ein fast vollständiges System mittelplatonischer Theologie. Im Mittelpunkt steht dabei die Lehre von den Dämonen; denn Apuleius will nachweisen, dass das Daimonion der persönliche Dämon (Schutzgeist) des Sokrates ist. Die Schrift stellt das ausführlichste und reichste Dokument zur Lehre der Mittelplatoniker von den Dämonen dar. Als Lehrvortrag ist es zugleich ein spektakuläres Zeugnis für die Redekunst des 2. Jahrhunderts n.Chr

Abdominal drainage after elective colorectal surgery: propensity score-matched retrospective analysis of an Italian cohort

background: In italy, surgeons continue to drain the abdominal cavity in more than 50 per cent of patients after colorectal resection. the aim of this study was to evaluate the impact of abdominal drain placement on early adverse events in patients undergoing elective colorectal surgery. methods: a database was retrospectively analysed through a 1:1 propensity score-matching model including 21 covariates. the primary endpoint was the postoperative duration of stay, and the secondary endpoints were surgical site infections, infectious morbidity rate defined as surgical site infections plus pulmonary infections plus urinary infections, anastomotic leakage, overall morbidity rate, major morbidity rate, reoperation and mortality rates. the results of multiple logistic regression analyses were presented as odds ratios (OR) and 95 per cent c.i. results: a total of 6157 patients were analysed to produce two well-balanced groups of 1802 patients: group (A), no abdominal drain(s) and group (B), abdominal drain(s). group a versus group B showed a significantly lower risk of postoperative duration of stay >6 days (OR 0.60; 95 per cent c.i. 0.51-0.70; P < 0.001). a mean postoperative duration of stay difference of 0.86 days was detected between groups. no difference was recorded between the two groups for all the other endpoints. conclusion: this study confirms that placement of abdominal drain(s) after elective colorectal surgery is associated with a non-clinically significant longer (0.86 days) postoperative duration of stay but has no impact on any other secondary outcomes, confirming that abdominal drains should not be used routinely in colorectal surgery

Bowel preparation for elective colorectal resection: multi-treatment machine learning analysis on 6241 cases from a prospective Italian cohort

background current evidence concerning bowel preparation before elective colorectal surgery is still controversial. this study aimed to compare the incidence of anastomotic leakage (AL), surgical site infections (SSIs), and overall morbidity (any adverse event, OM) after elective colorectal surgery using four different types of bowel preparation. methods a prospective database gathered among 78 Italian surgical centers in two prospective studies, including 6241 patients who underwent elective colorectal resection with anastomosis for malignant or benign disease, was re-analyzed through a multi-treatment machine-learning model considering no bowel preparation (NBP; No. = 3742; 60.0%) as the reference treatment arm, compared to oral antibiotics alone (oA; No. = 406; 6.5%), mechanical bowel preparation alone (MBP; No. = 1486; 23.8%), or in combination with oAB (MoABP; No. = 607; 9.7%). twenty covariates related to biometric data, surgical procedures, perioperative management, and hospital/center data potentially affecting outcomes were included and balanced into the model. the primary endpoints were AL, SSIs, and OM. all the results were reported as odds ratio (OR) with 95% confidence intervals (95% CI). results compared to NBP, MBP showed significantly higher AL risk (OR 1.82; 95% CI 1.23-2.71; p = .003) and OM risk (OR 1.38; 95% CI 1.10-1.72; p = .005), no significant differences for all the endpoints were recorded in the oA group, whereas MoABP showed a significantly reduced SSI risk (OR 0.45; 95% CI 0.25-0.79; p = .008). conclusions MoABP significantly reduced the SSI risk after elective colorectal surgery, therefore representing a valid alternative to NBP