Novel methods for chemiluminescent detection of reporter enzymes.

Chemiluminescent reporter gene assays provide highly sensitive, quantitative detection in simple, rapid assay formats for detection of reporter enzymes that are widely employed in gene expression studies. Chemiluminescent detection methodologies typically provide up to 100-1000x higher sensitivities than may be achieved with fluorescent or colorimetric enzyme substrates. The variety of chemiluminescent 1,2-dioxetane substrates available enable assay versatility, allowing optimization of assay formats with the available instrumentation, and are ideal for use in gene expression assays performed in both biomedical and pharmaceutical research. In addition, 1,2,-dioxetane chemistries can be multiplexed with luciferase detection reagents for dual detection of multiple enzymes in a single sample. These assays are amenable to automation with a broad range of instrumentation for high throughput compound screening.

RP 62203, a 5-hydroxytryptamine2 antagonist, enhances deep NREM sleep in rats.

RP 62203, a naphtosultam derivative, is an antagonist at the 5-hydroxytryptamine2 (HT2) receptor. The sleep pattern of rats treated orally with RP 62203 was studied at doses ranging from 0.5 to 4 mg/kg. Following RP 62203 administration, the duration of deep nonrapid eye movement (NREM) sleep was found to increase at the expense of wakefulness in a dose-dependent manner from 0.5 mg/kg. The 5-HT2 receptor agonist DOI and the 5-HT1a receptor agonist 8 OH-DPAT induced a dose-related increase in wakefulness; treatment with RP 62203 reversed the enhancement of wakefulness produced by DOI but not that produced by 8 OH-DPAT. These data provide further evidence for the involvement of 5-HT2 receptors in the regulation of NREM sleep in rats. RP 62203 could therefore be of clinical interest in the management of sleep disorders, particularly those developing within a psychiatric context.

Are 5-HT2 antagonists endowed with anxiolytic properties in rodents?

The precise role of serotonin (5-HT) in anxiety remains unclear. We report here on the effects of RP 62203, a new 5-HT2 antagonist, and ritanserin in different animal models of anxiety. In the elevated plus-maze in mice, RP 62203 increased dose-dependently the percentage of entries onto, and time spent on open arms, over the dose range 0.25-4 mg.kg-1 p.o. By contrast, ritanserin was ineffective up to the dose of 4 mg.kg-1 p.o. In addition, both compounds were tested against the anxiogenic compound FG 7142 (20 mg.kg-1, i.p.) in the plus-maze test in mice and via electrocorticographic recordings (ECoG) in rats. The anxiolytic effect of RP 62203 is antagonized by FG 7142 at a dose devoid of anxiogenic properties. A similar interaction between RP 62203 and FG 7142 is observed in ECoG studies. In contrast, ritanserin seemed to potentiate the anxiogenic and awakening activities of FG 7142. These results demonstrate that RP 62203, a selective 5-HT2 antagonist, possesses anxiolytic properties in rodents suggesting that 5-HT2 receptors are involved in the control of anxiety.

[Aortic paraprosthetic infection. Diagnostic and therapeutic difficulties. Apropos of a case]. / Infection paraprothétique aortique. Difficultés diagnostiques et thérapeutiques. A propos d'un cas.

Despite advances in vascular surgical techniques and antimicrobial therapy, aortic graft infection remains a difficult clinical problem to manage. We report a case of secondary paraprosthetic fistula. All complementary investigations were negative and the fistula was diagnosed by laparotomy. In situ replacement of the infected graft was decided despite signs of infection. The postoperative course was complicated by septic shock, successfully treated by organism-specific antibiotic therapy. In conclusion, complementary investigations should not delay laparotomy in patients with a aortic prosthesis presenting with unexplained infection. They must be considered to be suffering from a graft infection until proven otherwise. Conditions in situ replacement of the graft appears to be possible under these.

[Temporomandibular staphylococcal arthritis]. / Arthrite temporo-maxillaire à staphylocoque.

Two cases of temporo-mandibular joint infection with several blood cultures positive for Staphylococcus aureus are reported. Positive diagnosis depended on the importance of the pain and trismus, as well as the lack of abscess formation. The rapid development of ankylosis requires treatment to be started as soon as the first signs appear.

[Protein C deficiency and cerebral venous thrombosis in pregnancy]. / Déficit en protéine C et thrombose veineuse cérébrale au cours de la grossesse.

A fatal case of cerebral venous thrombosis associated with protein C deficiency is reported. It occurred at six months of gestation in a 25 year old patient during her first pregnancy. She had generalized seizures. Computed axial tomography displayed cerebral haemorrhagic infarction, and the carotid angiograms signs of superior longitudinal sinus thrombosis. Before the patient's death on the seventh day after admission, protein C deficiency was discovered. The occurrence of thrombosis of the superior longitudinal sinus during pregnancy, as well as that of protein C deficiency in atypical cases of thromboembolic disease, are discussed.

[Use of norepinephrine in the treatment of septic shock]. / Utilisation de la noradrénaline dans le traitement du choc septique.

The effects of noradrenaline were studied in 16 patients, with either a hyperkinetic septic shock syndrome or a septic shock resistant to dobutamine treatment. The study aimed to restore normal tissue perfusion pressure, assessed by a return to normal of urine output or blood pressure. An optimal left ventricular filling pressure, estimated by the pulmonary capillary wedge pressure, was obtained for each patient using a Swan-Ganz catheter. The administration of 10.6 +/- 0.5 micrograms.kg-1.min-1 dobutamine (starting dose: 6 micrograms.kg-1.min-1) was started when the cardiac index (CI) was less than 3.3 l.min-1.m-2 after vascular filling with plasma expanders. Patients became eligible for noradrenaline treatment when they fulfilled the following conditions: arterial systolic pressure (Pasys) less than or equal to 90 mmHg; systemic vascular resistances less than or equal to 600 dyn.s.cm-5; CI greater than 3.5 l.min-1.m-2; persistent oliguria (less than 30 ml.h-1). This drug was given at a constant rate with a starting dose of 0.5 micrograms.kg-1.min-1, increased every 10 min by 0.3 to 0.6 micrograms.kg-1.min-1 according to the effects on Pasys and hourly urine output. Eight patients received noradrenaline alone; the efficient dose was 0.9 +/- 0.2 micrograms.kg-1.min-1, and it was used for a mean 5.1 +/- 1 days. CI increased in those patients who were given both noradrenaline and dobutamine. Thirteen out of the 16 patients had a dramatic increase in urine output; only three patients remained oliguric. There were no effects on serum creatinine concentration, anion gap, intrapulmonary shunt and oxygen consumption.(ABSTRACT TRUNCATED AT 250 WORDS)

[Evaluation of anesthetic activity in the operating room. Use of beta relative cost index]. / Evaluation de l'activité anesthésique au bloc opératoire. Utilisation de l'ICR bêta.

OBJECTIVE: To evaluate the anaesthetic activity in the operating rooms using the newly reconstructed RCI beta, or relative cost index beta, a specific tool for analysis of anaesthetic activity and the linked cost. STUDY DESIGN: Prospective multicentric survey. METHODS: All scheduled anaesthetic procedures performed in March 1995 were collected. RCI beta items were entered in a standardized data base. Gender, age group, time of admission to the operating room, time of incision, time of exit from the operating room, and the code number of the surgical or radiological procedure were added on request of the steering committee. RESULTS: Complete responses were obtained from 35 out of the 37 contacted departments. A total of 31,391 procedures were analysed. Only 14% of patients were of ASA class over 2. Anaesthetic practices were comparable between institutions. Only the incidence of special circumstances and techniques was higher in University hospitals. General anaesthesia was the most widely used technique (76.58%). A large proportion (19%) of anaesthetics were given for endoscopy and radiology. More than 87% of patients were monitored postoperatively in recovery areas. There was a lower correlation between the theoretical standard duration and median actual duration of the perioperative period than the operative period (r = 0.54 vs r = 0.81). DISCUSSION: Part of our activity could be described with RCI beta. The obtained data allow a comparison of anaesthetic activity in the operating rooms of different hospitals, departments and units. Further analysis of these data will also provide information about the types of surgical procedures and the level of global activity.

[A scale of perioperative satisfaction for anesthesia. I--Construction and validation]. / Echelle de vécu périopératoire de l'anesthésie. I--construction et validation.

OBJECTIVE: To develop and to validate a scale assessing perioperative patient's satisfaction with anaesthesia (Evan). STUDY DESIGN: Descriptive and evaluative study. PATIENTS: The study included 742 adults undergoing a surgical or a diagnostic procedure under general anaesthesia. Emergency, ambulatory and obstetrical cases were excluded. METHODS: A multidisciplinary working party produced 85 questions focusing on various pertinent areas describing satisfaction. After a validation, 25 out of them were selected for the questionnaire. The latter was completed within the 24 hours following anaesthesia by 742 inpatients. RESULTS: Item analysis showed a homogeneous distribution of the answers to each item. Main component analysis allowed to explain 53% of total variance. Six dimensions were isolated by the exploratory analysis: anxiety, embarrassment, fear, pain-discomfort, information and physical needs. Scoring method followed a simple additive model: for each dimension, the scale scored 0-100. The global score represented the sum of the six dimensions also scored 0-100. Acceptability of Evan questionnaire was satisfactory, with a spontaneous non response rate of less than 1% and a completion duration at 11 +/- 8 min. CONCLUSION: A self-completed questionnaire on patient's satisfaction with anaesthetic period was validated, allowing a global and multidimensional assessment of patient's satisfaction.

[A scale of perioperative satisfaction for anesthesia. II--Preliminary results]. / Echelle de vécu périopératoire de l'anesthésie. II--Résultats préliminaires.

PURPOSE: To assess the patient's experience of anaesthesia in the early postoperative period, with a self-completed questionnaire (Evan). STUDY DESIGN: Descriptive and evaluative study. PATIENTS: The study included 742 adults undergoing an elective surgical or non surgical procedure under anaesthesia. METHODS: An Evan questionnaire with 25 questions was completed 24 hours after anaesthesia by the patient. The questionnaire explored six areas, each one being marked out from 0 to 100, as the visual analogue scale. The marks were compared with consideration of age, gender, ASA physical class, type of anaesthesia, anaesthesia duration and type of surgery. RESULTS: The mean global mark was 76 +/- 9 (min-max: 34-99). Marks were lower in the youngest patients, in females, in ASA 1 patients, in longest surgical procedures, especially with regard to areas belonging to "apprehension", "pain-discomfort" and "physical needs". The lowest mark was given for the "information" provided during the pre-anaesthetic evaluation. Differences in marks occurred also between surgical specialities. CONCLUSION: The Evan questionnaire is a valuable tool for assessing the patient's opinion on the perioperative period. Further studies are required to extend its use to other fields, as ambulatory surgery.

[Ischemic heart failure making the diagnosis of a Vaquez disease: a rare event]. / Choc cardiogénique ischémique révélateur d'une maladie de Vaquez, un événement rare.

We present the case of a 46-year-old patient without any past medical history, admitted to our ICU for cardiogenic shock complicating acute coronary syndrome. The blood tests found polycethemia, a polycethemia vera was suspected and confirmed by genetic analysis. Ischemic heart failure as an initial symptom of polycethemia vera and its treatment by arterial bleeding is a rare event that we describe in this article.

[Perioperative conflicts between anaesthesiologists and surgeons: ethics and professionalism]. / Conflits en période périopératoire: un enjeu collectif, éthique et professionnel.

In the perioperative period, several potential conflicts between anaesthetists/intensive care specialists and surgeons may exist. They are detrimental to the quality of patient care and to the well-being of the teams. They are a source of medical errors and contribute to burn-out. Patients can become the victims of such conflicts, which deserve ethical reflection. Their resolution through analysis and shared solutions is necessary. This article seeks to analyse these conflicts, taking into account their specificities and constraints. In order to understand this context, it is important to consider the specificities of each group involved and the records of such situations. Several factors can prevent these conflicts, first and foremost the patients themselves and the quality of the care that is provided. Medical deontology aims mainly at preventing and resolving these conflicts. Generally speaking, the quality approach which is increasingly applied in health care institutions (involving declarations of adverse events, morbidity/mortality reviews, benchmarking, analysis and improvement of practices, etc.) also contributes to the prevention and resolution of disagreements. The teaching of communication techniques that begins with the initial training, the evaluation of team behaviours (through simulation training for example), the respect of others' constraints, particularly when it comes to learning, as well as transparency regarding conflicts of interests, are all additional elements of conflict prevention. Lastly, conflicts may at times be caused by deviant behaviours, which must be met with a clear and uncompromising collective and institutional approach. This article concludes by offering a standardised approach for conflict resolution.

Critical appraisal of organ procurement under Maastricht 3 condition.

The ethics committee of the French Society of Anesthesia and Intensive Care (Sfar) has been requested by the French Biomedical Agency to consider the issue of organ donation in patients after the decision to withdraw life-supportive therapies has been taken. This type of organ donation is performed in the USA, Canada, the United Kingdom, the Netherlands and Belgium. The three former countries have published recommendations formalizing procedures and operations. The French Society of Anesthesia and Intensive Care (Société française d'anesthésie et de reanimation [Sfar]) ethics committee has considered this issue and envisioned the different aspects of the whole process. Consequently, it sounded a note of caution regarding the applicability of this type of organ procurement in unselected patients following a decision to withdraw life-supportive therapies. According to French regulations concerning organ procurement in brain-dead patients, the committee stresses the need to restrict this specific way of procurement to severely brain-injured patients, once confirmatory investigations predicting a catastrophic prognosis have been performed. This suggests that the nature of the confirmatory investigation required should be formalized by the French Biomedical Agency on behalf of the French parliamentarians, which should help preserve population trust regarding organ procurement and provide a framework for medical decision. This text has been endorsed by the Sfar.

[Implementation of morbidity and mortality conferences in French intensive care units: a survey]. / Enquête nationale sur la pratique des "Revues de morbi-mortalité" en réanimation.

OBJECTIVE: A national survey was conducted by the "Collège français d'anesthésie et de réanimation (CFAR)" and the "Collège des bonnes pratiques en réanimation (CBPR)", to analyze the implementation of morbidity and mortality conferences (MMCs) in French intensive care units (ICUs). STUDY DESIGN: An electronic questionnaire was set up. We directed the survey at French ICUs physicians registered in the two Colleges directories, only one form was filled in by each participating unit. RESULTS: From December 2009 to February 2010, Among the 170 replies, 120 ICUs (71%) practiced MMC. No difference in the typology of the two groups was found. The median annual number of MMCs was 4 per year (1-15). The perimeter of the MMCs concerned only the ICU unit in 70 cases (58%), more than one ICU unit in the same department in 11 cases (9.8%), more than one department of ICU in 16 cases (13%) and other departments in 57 cases (48%). The events analyzed were: all deaths in 45 cases (37.5%), unexpected deaths in 50 cases (41.7%), severe adverse events in 67 cases (55.8%) and other events in 19 cases (15.8%). At least one adverse event defined by the two colleges in the process of "accreditation" was analyzed in 86 cases (72%). Participation of a physician of at least one other unit was reported in 56 cases (47%) and of medical students in 62 cases (52%). The low rate of participation of ICU nurses was reported in 62 cases (69.2%) and their absence in 35 cases (29%). MMCs consequences were drafting of new procedure in 99 cases (83%), changes in procedures in 75 cases (63%), conducting training programs in 60 cases (50%), organizational changes in 86 cases (72%), adverse event declaration in 21 cases (18%) and monitoring indicators in 40 cases (33%). Among units which did not practice MMCs, Identified obstacles were organizational causes in 25 cases (50%), inexperience in seven cases (14%), lack of methodology in 4 cases (8%), realization of other methods of formative assessment in 4 cases (8%) and physician's refusal in three cases (6%). The fear of medico-legal problem was never reported as a barrier to MMCs practice. Forty-five units (90%) projected to practice MMR. CONCLUSION: This survey showed that the practice of MMR is common in French ICUs, allowing the identification of organizational problems, but also of training needs, joining one of the initial concerns that have led to their implementation. Expanding the participation to non-physician members of the units should be encouraged, without underestimating the difficulties particularly in the organizational domains that represent an obstacle to development of MMCs.

[Critical appraisal of organ procurement under Maastricht 3 condition]. / Analyse critique du prélèvement en condition M3 de Maastricht.

The ethics committee of the French Society of Anesthesia and Intensive Care (Sfar) has been requested by the French Biomedical Agency to consider the issue of organ donation in patients after a decision of withdrawing life supporting therapies has been taken. This category of organ donation is performed in the USA, Canada, United Kingdom, the Netherlands and Belgium. The three former countries have published recommendations, which formalize procedures and operations. The Sfar ethics committee has considered this issue and envisioned the different aspects of the whole process. Consequently, it sounds a note of caution regarding the applicability of this type of organ procurement in unselected patient following a decision to withdraw life supporting therapies. According to the French regulation concerning organ procurement in brain dead patients, the committee stresses the need to restrict this specific way of procurement to severely brain injured patients, once confirmatory investigations predicting a catastrophic prognosis have been performed. It suggests that the nature of the confirmatory investigation required should be formalized by the French Biomedical Agency on behalf of the French parliamentarians. This should help preserving population trust regarding organ procurement and provide a framework to medical decision. This text has been endorsed by the Sfar.

[Euthanasia, assisted suicide and palliative care: a review by the Ethics Committee of the French Society of Anaesthesia and Intensive Care]. / Fin de vie, euthanasie et suicide assisté : une mise au point de la Société française d'anesthésie et de réanimation (Sfar).

CONTEXT: Management of the end of life is a major social issue which was addressed in France by law, on April 22nd 2005. Nevertheless, a debate has emerged within French society about the legalization of euthanasia and/or assisted suicide (E/AS). This issue raises questions for doctors and most especially for anesthetists and intensive care physicians. OBJECTIVE: To highlight, dispassionately and without dogmatism, key points taken from the published literature and the experience of countries which have legislated for E/AS. RESULTS: The current French law addresses most of the end of life issues an intensive care physician might encounter. It is credited for imposing palliative care when therapies have become senseless and are withdrawn. However, this requirement for palliative care is generally applied too late in the course of a fatal illness. There is a great need for more education and stronger incentives for early action in this area. On the rare occasions when E/AS is requested, either by the patient or their loved-ones, it often results from a failure to consider that treatments have become senseless and conflict with patient's best interest. The implementation of E/AS cannot be reduced to a simple affirmation of the Principle of autonomy. Such procedures present genuine difficulties and the risk of drift. CONCLUSION: We deliver a message of prudence and caution. Should we address painful end of life and moral suffering issues, by suppressing the subject, i.e. ending the patient's life, when comprehensive palliative care has not first been fully granted to all patients in need of it ?