A new agarose-based microsystem to investigate cell response to prolonged confinement.

Emerging evidence suggests the importance of mechanical stimuli in normal and pathological situations for the control of many critical cellular functions. While the effect of matrix stiffness has been and is still extensively studied, few studies have focused on the role of mechanical stresses. The main limitation of such analyses is the lack of standard in vitro assays enabling extended mechanical stimulation compatible with dynamic biological and biophysical cell characterization. We have developed an agarose-based microsystem, the soft cell confiner, which enables the precise control of confinement for single or mixed cell populations. The rigidity of the confiner matches physiological conditions and its porosity enables passive medium renewal. It is compatible with time-lapse microscopy, in situ immunostaining, and standard molecular analyses, and can be used with both adherent and non-adherent cell lines. Cell proliferation of various cell lines (hematopoietic cells, MCF10A epithelial breast cells and HS27A stromal cells) was followed for several days up to confluence using video-microscopy and further documented by Western blot and immunostaining. Interestingly, even though the nuclear projected area was much larger upon confinement, with many highly deformed nuclei (non-circular shape), cell viability, assessed by live and dead cell staining, was unaffected for up to 8 days in the confiner. However, there was a decrease in cell proliferation upon confinement for all cell lines tested. The soft cell confiner is thus a valuable tool to decipher the effects of long-term confinement and deformation on the biology of cell populations. This tool will be instrumental in deciphering the impact of nuclear and cytoskeletal mechanosensitivity in normal and pathological conditions involving highly confined situations, such as those reported upon aging with fibrosis or during cancer.

SU-D-213CD-06: Workflow and Safety Systems of a Linac-MR Sim-Brachytherapy MRgRT™ Facility.

PURPOSE: To develop the operational workflow and safety systems of a magnetic resonance-guided radiotherapy system (MRgRT™), which comprises an MR scanner on rails that travels between a linac vault, MR simulation room and brachytherapy suite. METHODS: To develop a safe and streamlined clinical workflow, we conducted a comprehensive process review based on a layered approach to overall MRgRT safety that included i) facility design, (ii) workflow iii) system design and interlocks and iv) policies and procedures. We applied existing guidelines for MR and radiation safety, and employed system-level failure modes and effects analyses to design the MRgRT facility and clinical procedures. RESULTS: In the MRgRT system configuration, the MR and treatment systems are physically decoupled and used independently requiring novel administration of existing MR and radiation guidelines. A key element for the safe operation of the moving MR unit is the concept that all three rooms represent zone 4 areas (American College of Radiology guidelines). Using this concept, we applied MR guidelines to develop safe procedures for the overall suite, including screening of all persons entering the suite in zone 2 and control of ferromagnetic materials. We generated a clinical workflow that ensures expedient and safe transition between MR imaging and treatment delivery in both the linac and brachytherapy rooms. In addition, we designed emergency protocols for MRgRT, which helped drive requirements for the facility and system design, e.g., need for an accessible MR-safe stretcher. CONCLUSIONS: We designed the first comprehensive description of the MRgRT workflow, interlocking systems and safety procedures. With this layered approach to safety, we addressed critical aspects regarding safe operation and workflow for the system and provided multiple redundancies for key processes. Coupled with customized staff training, the proposed design ensures the safe operation of the MRgRT facility. This work has received research personnel support from IMRIS.

Effect of operating conditions on ochratoxin A extraction from roasted coffee.

Operating conditions affect ochratoxin A (OTA) extraction from roasted coffee. The OTA content found in the beverage can thus be greater than that found in the roasted coffee used to prepare it. Three extraction parameters were studied for roasted coffee: type of extraction solvent (alkaline, neutral, acid), temperature (ambient temperature/23 degrees C, 60 degrees C and 85 degrees C), and extraction time (5, 20, 30, 40, 50, 60 and 80 min). The alkaline solvent used in the method recommended by the European Union extracted OTA better, but a maximum content was obtained at 60 degrees C after 50 min. At least a 100% improvement in extraction was obtained when compared with the European Union usual quantification method that is carried out at ambient temperature. It turned out that the OTA extraction parameters for roasted coffee, as defined by that method, were not optimum and needed to be modified. These results were verified in double-extraction experiments showing that OTA is not completely extracted by this method. Confirmation was obtained by comparison of extraction methods on several commercial samples of roasted coffee.

[Postoperative pain after hysterectomy through vaginal routes using electro surgical bipolar vessel sealing versus conventional suture ligature]. / Douleur postopératoire après hystérectomie par voie vaginale selon la méthode d'hémostase utilisée: thermofusion ou suture aux fils.

OBJECTIVES: The purpose of the study was to compare the postoperative pain of patients who had a hysterectomy through vaginal route according to the process of binding: wire or electrosurgical bipolar vessel sealing. PATIENTS AND METHODS: Retrospective study carried out in the 60 last patients who underwent a hysterectomy by vaginal route for a benign pathology in the gynaecological service of surgery of the CHI Poissy-Saint-Germain-en-Laye until March 2006. Among these patients, 32 had profited from a binding by wire and 28 of the electrosurgical bipolar vessel sealing. The studied criteria were the post-operative pain, total morphine consumption and the durations of the analgesic treatment, the hospitalisation and intervention time. RESULTS: The postoperative pain in the first 24 hours was twice lower using thermofusion; it was valid in immediate post-operative period and after 24 hours. In addition, total morphine consumption was also significantly lower using thermofusion. DISCUSSION AND CONCLUSION: This pilot study shows that the electrosurgical bipolar vessel sealing allows a reduction in the pain into the immediate postoperative period. Other prospective and randomised studies would allow it and conclude on the duration of hospitalisation, the quality of life from the patients and the cost in terms of public health.

[Interest of modern imagery for conservative management of a placenta percreta]. / Prise en charge conservatrice d'un placenta percreta. Apport des nouvelles techniques d'imagerie.

The placenta percreta is a rare form of anomaly of placental insertion threatening the maternal and foetal lives. The incidence of abnormal placental adhesion is correlated to frequency of caesarean sections and advanced maternal age. Patients who are at high risk should be identified during pregnancy by ultrasound examination with Color Doppler looking for characteristic features. The practice of MRI in case of echographic suspicion may be useful, particularly in posterior placentas. A late diagnosis, in an emergency context, leads generally to hysterectomy and even to partial resection of neighbour organs. Thus, we report the case of a conservative management associating uterine embolisation following an elective caesarean delivery at 36 weeks gestation. In postpartum, the placental involution was followed clinically and by imagery. In our case, a total abdominal hysterectomy was performed on the fifth postoperative week because of a severe antibiotics resistant infection. No blood transfusion was required and the postoperative period was uneventful. This clinical case aims to show, through an analysis of the recent data of the literature, the interest of modern imagery to select patients with suspected placenta percreta that would be suitable candidates for conservative management.

[Proposition of a simplified investigation model for adverse outcomes in obstetrics and gynaecology]. / Proposition d'un modèle d'analyse simplifiée d'événements indésirables en chirurgie gynécologique et en obstétrique.

Investigation of adverse outcomes in obstetrics and gynaecology requires a systematic and systemically oriented approach. The main objective of these assessments is to decrease their incidence. Lethal complications in obstetrics and gynaecological surgery have become so rare in developed countries that we have to grapple mainly with severe morbidities. A model for such investigation is presented, based on answers to 3 questions: What? How? and Why? These questions correspond to 3 components of quality assurance: Outcome, Process, and Structure, respectively. Aspects of Reason's model of accidents are also included, in the form of "active failures" and "latent conditions." The latter are divided into basic contributory factors, 4 of which are derived from a model of human factors influencing behaviour in anaesthesia or flight-crew environments. To illustrate its basic nature, the model is applied to anonymized cases of surgical complications. Application of the model to the reactive investigation of close calls is discussed, as is application to the proactive interrogation of the care system for the presence and nature of hazards that pose threats to safety.

[Use of prosthetic materials in reconstructive pelvic floor surgery. An evidence-based analysis]. / Utilisation de matériaux prothétiques dans la chirurgie réparatrice des prolapsus pelviens. Analyse factuelle des connaissances.

Many surgical procedures for the repair of pelvic organ prolapse are used nowadays. Reconstructive pelvic surgery continues to evolve while surgeons continue to search the definitive surgical cure and have to choose the most appropriate procedures for their patients. Concerning the vaginal approach procedures, there is an increasing interest in the use of synthetic meshes which are at present widely used for surgical repair of pelvic organ prolapse. Prosthetic repair seems to be more reliable, especially when native tissues are of poor quality. The use of synthetic meshes may also simplify surgical procedures and reduce operative duration and morbidity. Material must be inert, permanent and resistant to infection. Based on authors' and other researchers' published experimental and clinical experience, polypropylene is assumed to be the most appropriate material for the vaginal repair of pelvic organ prolapse. However, since no standardized outcome measure is available, it is difficult to compare the results of surgical procedures. Only in recent studies, the subjective cure rates (patient satisfaction and outcome) have been assessed as well as the objective cure rates determined by the investigators. The subjective cure rate is probably more influenced by the functional outcome and sexual activity than by the anatomical result. Continuous evaluation is necessary to study replacement synthetic materials which should improve the rate of prolapse recurrence and reduce the risk of complications. Randomized controlled trials are required to determine which surgical procedures and type of prosthesis are most suitable. This review evaluates the properties of prosthetic materials, their complications and the most common procedures involved in the use of meshes for pelvic reconstructive surgery.

[Raloxifene and breast: from the SERMs concept to its place in clinical practice]. / Raloxifène et tissu mammaire : du concept des SERMs à la place de ce composé en pratique clinique.

Raloxifene is a second generation Selective Estrogen Receptor Modulator. It is indicated in prevention and treatment of post-menopausal osteoporosis. This drug has an estrogen agonistic effect on bone tissue, an estrogen-antagonistic effect in breast and endometrial tissue. It inhibits estrogen induced breast cells proliferation and, in animal studies, it prevents the growth of chemically induced mammary tumors. Raloxifene has a limited effectiveness in women with advanced breast cancers. The effects of raloxifene after treated breast cancer and sequential use of tamoxifene after raloxifene (or the contrary) are not known and should be tested in specific pre-clinical and clinical studies. In osteoporotic women included in the MORE trial, the risk reduction of hormone-dependent breast cancer due to raloxifene raises the issue of some effectiveness of raloxifene in breast cancer prevention. The STAR trial is currently comparing tamoxifene vs raloxifene in breast cancer prevention in women at increased risk of breast cancer. Like tamoxifene, raloxifene increases the risk of venous thrombo-embolic events. On the other hand, in a subgroup of women at increased vascular risk in the MORE trial, the reduction of cardiovascular events raises the issue of an effect of raloxifene in the prevention of coronary events. The ongoing RUTH trial is testing this assumption and it is comparing raloxifene to a placebo in women at increased vascular risk. Today, in clinical practice, before the results of ongoing trail, raloxifene should be used in the setting of osteoporosis prevention and treatment.

[Ovarian choriocarcinoma: about one case]. / Choriocarcinome ovarien: à propos d'un cas.

We report the case of a woman, 43 years old, presenting with an ovarian choriocarcinoma. This patient, addressed for a right-side suspicious pelvic mass, had an exploratory laparotomy. Frozen section found an ovarian cancer. The patient had total hysterectomy, left annexectomy and a complete staging surgery. Histology concluded to an ovarian gestational choriocarcinoma. It is a rare germinal tumour, the treatment of which was medical, with radical exeresis and complete staging surgery associated with polychemotherapy.

Impact of the Coffea canephora gene introgression on beverage quality of C. arabica.

Lines of Coffea arabica derived from the Timor Hybrid (hybrid between C. arabica and C. canephora) are resistant to coffee leaf rust (Hemileia vastatrix) and to the nematode Meloidogyne exigua. The introgression of C. canephora resistance genes is suspected of causing a drop in beverage quality. Coffee samples from pure lines, compared in a Trial 1, and from F1 hybrids and parental lines from a half-diallel trial in a Trial 2, were studied for beverage quality, chemical composition and amount of introgressed genetic material. Chemical analyses (caffeine, chlorogenic acids, fat, trigonelline, sucrose) were carried out with near-infrared spectrometry by reflectance of green coffee. The number of amplified fragment length polymorphic (AFLP) markers introgressed from the Timor Hybrid varied from 1 to 37 for the lines studied. There were significant differences between lines for all of the biochemical compounds analysed and for the acidity and the overall standard of the beverage. Two lines (T17927, T17924) were significantly poorer than the controls for sucrose and beverage acidity. T17924 also had more chlorogenic acids and was poorer for the overall standard. However, two highly introgressed lines, T17934 and T17931 (25 and 30 AFLP markers, respectively), did not differ from the non-introgressed controls. There were no correlations between the number of AFLP markers and the chemical contents or beverage attributes. Significant correlations were found between the performance of the parents and their general combining ability for beverage quality. It was concluded that it should be possible to find lines with both the desired resistance genes and good beverage quality. Selection can avoid accompanying the introgression of resistance genes with a drop in beverage quality.

Characterization of promoter elements directing Mona/Gads molecular adapter expression in T and myelomonocytic cells: involvement of the AML-1 transcription factor.

Monocytic adaptor (Mona, also called Gads) is a molecular adaptor implicated in T cell activation and macrophage differentiation. The objective of this study was to identify elements regulating specific expression of Mona/Gads in human T cell and myelomonocytic cell lines. We first confirmed that the -2000 to +150 genomic region relative to the Mona gene transcription start site is sufficient to direct specific reporter gene expression in T cell lines, Jurkat, and MOLT-4 and in the immature myeloid cell lines, KG1a and RC2A. Deletion analysis and electrophoresis mobility shift assay identified several cis regulatory elements: overlapping initiator sequences, one interferon response factor-2 (IRF-2)-binding site at position -154, one GC box recognized by Sp1 and Sp3 at position -52, and two acute myeloid leukemia (AML)-1 binding sites at positions -70 and -13. Site-directed mutagenesis experiments indicated a key role of AML-1 for driving Mona expression in T cells and myeloid cells, and involvement of Sp1/Sp3 and IRF-2 transcription factors to modulate Mona expression in a cell-specific manner.

Genomic organization and restricted expression of the human Mona/Gads gene suggests regulation by two specific promoters.

Monocytic adaptor (Mona) also known as Gads is a Grb2-related adaptor whose expression is restricted to hematopoietic cells. It plays an important role in intracellular signaling in T cells, monocytic cells, and platelets. Here we investigated the regulatory aspects of Mona expression in human hematopoietic cells. This was carried out by combining nucleotide sequence analyzes and experimental approaches. We confirmed that Mona expression is restricted to T-cell, myeloid and platelet lineages. In the various cells examined, we detected two major Mona transcripts (1.9 and 4 kb), likely resulting from the alternative use of two polyadenylation sites. Consequently, Mona transcripts of the same size have identical 3' untranslated region (UTR), irrespective of the cell type. In contrast, Mona transcripts contain either 5' UTR-1A or -1B exons, that were detected in a cell-lineage specific manner. Thus, T cells and several myeloid cell lines express 5' UTR-1A-containing transcripts, whereas platelets and cell lines exhibiting megakaryocytic potential express 5' UTR-1B-containing transcripts. Interestingly, 5' UTR-1A is generated from an exon located approximately 45 kb upstream of exon 1B. This suggested that lineage-restricted transcription of the Mona gene is controlled by specific promoters. Indeed, 2-kb genomic fragments upstream of each 5'-UTR showed lineage-restricted ability to drive expression of luc reporter gene.

Fate of mucilage cell wall polysaccharides during coffee fermentation.

Effects of a 20-h fermentation on cell wall polysaccharides from the mucilage of pulped coffee beans were examined and compared to those of unfermented beans, on alcohol insoluble residues (AIRs), their hot-water-soluble crude pectic substances (PECTs), and their hot-water-insoluble residues (RESs). Yields and compositions were very similar: AIRs, which consisted of approximately 30% highly methylated pectic substances, approximately 9% cellulose, and approximately 15% neutral noncellulosic polysaccharides, exhibited no apparent degradation. However, PECTs from fermented beans were shown to have undergone a slight reduction of their intrinsic viscosity and weight-average molecular weight by capillary viscosimetry and high-performance size-exclusion chromatography. After fermentation, hot-water-insoluble pectic substances of RES exhibited partial de-esterification. Removal of coffee bean mucilage by natural fermentation seems to result from a restricted pectolysis, the mechanism of which remains to be elucidated.

Microbiological and biochemical study of coffee fermentation.

The coffee fermentation microflora were rich and mainly constituted of aerobic Gram-negative bacilli, with Erwinia and Klebsiella genuses at the highest frequencies. The best population increase was observed with lactic acid bacteria and yeasts, whereas those microorganisms that counted on a pectin medium remained constant during the fermentation step. Qualitatively, lactic acid bacteria belonged mainly to Leuconostoc mesenteroides species but the others microflora were relatively heterogeneous. The microorganisms isolated on pectin medium were Enterobacteriaceae, identified as Erwinia herbicola and Klebsiella pneumoniae, not reported as strong pectolytic strains. Throughout coffee fermentation, 60% of the simple sugars were degraded by the total microflora and not specifically by pectolytic microorganisms.

[Nancy medicines for hospices and prisons in 1805]. / Médicaments pour hospices et prisons en 1805 à Nancy.

The chemist François Mandel, past author of Pharmcopée de Nancy, writes a list of medicines for civil hospices and a formulary for prisons. To take care of these adult, poor sick, drugs and medicines are chosen simple, at little expense, but adequate to treat everyday diseases. The prison formulary holds many novel preparations. The two manuscripts have never been published.