Improvement of yeast biochip sensitivity using multilayer inorganic sol-gel substrates.

Today, microarray fluorescence detection is still limited because a great proportion of hybrids remain undetectable. In this paper we describe sol-gel optical multilayers (stacks of low- and high-index layers) deposited on glass slides which increase the fluorescence of DNA microarrays and favour the detection of fluorescent targets. An alternative to the expensive and time-consuming physical vapour deposition technology is proposed. It is a low-cost sol-gel coating of glass slides, each layer being made by "dipping" (alternatively in SiO2 or TiO2 solutions), "draining and drying". After the selection of the best surface layer of the substrates, the multilayer mirrors modelled for one (Cy3) or two (Cy3 and Cy5) fluorophores are spotted with a series of Yeast probes and compared to similar microarrays on standard glass slides through hybridisation experiments. The fluorescence images of the mirrors show increased signals for all the probes. The enhancement factors determined for Cy3 and for Cy3/Cy5 mirrors (10-12 and 4-5, respectively) are consistent with the initial modelling. This allows the assessment of the basal expression levels of Yeast low-expressed genes. Moreover, these substrates show a noticeable increase in sensitivity for induction/repression ratio measurements in differential gene expression experiments. So, they could be considered as promising tools for the analysis of small biological samples.

The restoration of Loch Leven, Scotland, UK.

This paper reviews the progress made towards the restoration of Loch Leven, the largest lake in lowland Scotland, over the last 20 years. In particular, the importance of direct regulation and of setting water quality objectives and targets is examined. Various means of engaging with stakeholders and the general public are also considered. Success criteria and catchment management initiatives are described and briefly reviewed.

Determination of air-to-water partition coefficients using automated multiple headspace extractions.

Air-to-water partition coefficients are experimentally determined using a multiple headspace extraction procedure and an automated headspace cell. The approach is first validated with 2-butanone and then applied to a homologous series of methyl ketones. As adsorptions of the most hydrophobic compounds occurred in the sampling cell, technical improvements have been tested. This study represents the first attempt to overcome analyte adsorptions by studying and minimizing the effect of the cell's adsorption of hydrophobic analytes on the determination of their partition coefficients. The present method allows the measurement of several analytes at the same time, in the ppm range, without calibration, and with a limited manpower.

Automated multiple headspace extraction procedure: adsorption modeling and determination of air-to-water partition coefficients.

The present work makes the first attempt to take into account adsorptions in the determination of partition coefficients by modeling the multiple headspace extraction (MHE) process. Modeling a six-step MHE procedure of a homologous series of methyl ketones revealed that their adsorption-desorption on the walls was the rate-limiting step. Moreover, a comparison between experimental and predicted MHE plots shows that only the last MHE points were affected by adsorption phenomena. Using cell materials with the lowest sorptive properties, partition coefficients were then accurately calculated from the first four MHE steps. This kinetic approach supports previous work in which adsorptions were lowered owing to the choice of sampling cell materials. It also justifies some reproducibility limitations of the MHE quantification procedure mentioned in the literature.

Focused microwave-assisted extraction of cocaine and benzoylecgonine from coca leaves.

Extraction of cocaine and benzoylecgonine from coca leaves was performed by focused microwave-assisted extraction (FMAE). Cocaine extraction was optimised with respect to the nature of the extracting solvent, the particle size distribution, the moisture of the sample, the applied microwave power and the radiation time. A central composite design was used to optimise the two latter parameters and to assess the robustness of the extraction method around the best conditions. FMAE generated extracts similar to those obtained by conventional solid-liquid extraction but in a more efficient manner, i.e. 30 s were sufficient to extract cocaine quantitatively from leaves. Analyses of cocaine and benzoylecgonine in coca leaves was carried out by capillary GC-FID and GC-MS for peak identification, as well as by capillary electrophoresis with UV detection.

Experimental design in supercritical fluid extraction of cocaine from coca leaves.

An optimisation procedure for the supercritical fluid extraction (SFE) of cocaine from the leaves of Erythroxylum coca var. coca was investigated by means of experimental design. After preliminary experiments where the SFE rate-controlling mechanism was determined, a central composite design was applied to evaluate interactions between selected SFE factors such as pressure, temperature, nature and percentage of the polar modifier, as well as to optimise these factors. Predicted and experimental contents of cocaine were compared and robustness of the extraction method estimated by drawing response surfaces. The analysis of cocaine in crude extracts was carried out by capillary GC equipped with a flame ionisation detector (GC-FID), as well as by capillary GC coupled with a mass spectrometer (GC-MS) for peak identification.

Peritoneal carcinomatosis from non-gynecologic malignancies: results of the EVOCAPE 1 multicentric prospective study.

BACKGROUND: Peritoneal carcinomatosis (PC) is a common evolution of digestive cancer, associated with a poor prognosis. Yet it is poorly documented in the literature. METHODS: Three hundred seventy patients with PC from non-gynecologic malignancies were followed prospectively: the PC was of gastric origin in 125 cases, of colorectal origin in 118 cases, of pancreatic origin in 58 cases, of unknown origin in 43 cases, and of miscellaneous origins in 26 cases. A previously reported PC staging system was used to classify these 370 patients. RESULTS: Mean and median overall survival periods were 6.0 and 3.1 months, respectively. Survival rates were mainly affected by the initial PC stage (9.8 months for Stage I with malignant peritoneal granulations less than 5 mm in greatest dimension, versus 3.7 months for Stage IV with large, malignant peritoneal masses more than 2 cm in greatest dimension). The presence of ascites was associated with poor survival of patients with gastric or pancreatic carcinoma. Differentiation of the primary tumor did not influence the prognoses of patients with PC. CONCLUSIONS: A better knowledge of the natural history of PC is needed, in view of the many Phase I, II, and III trials currently being conducted to evaluate aggressive multimodal therapeutic approaches to treating patients with PC from non-gynecologic malignancies.

Prevention of lymphorrhea by means of fibrin glue after axillary lymphadenectomy in breast cancer: prospective randomized trial.

A prospective randomized trial was carried out to evaluate the efficacy of fibrin glue in preventing lymphorrhea after axillary lymphadenectomy in breast cancer. One hundred and eight breast cancer patients, operated on by two senior surgeons, were randomized into two groups: group 1 (n = 58) without fibrin glue and group 2 (n = 50) with 2 ml of fibrin glue applied to the axillary dissection area at the end of the lymphadenectomy procedure. Early postoperative morbidity was 2/58 and 0/50 in groups 1 and 2, respectively. Mean daily postoperative drainage was significantly greater in group 1. The mean cumulative drainage quantity 6 days after the operation was 407.8 ml and 214.4 ml in groups 1 and 2, respectively (p = 0.001). The mean postoperative hospital stay was 10.1 days and 8.0 days in groups 1 and 2, respectively (p = 0.006). One delayed seroma was observed in each group. Fibrin glue seems to reduce daily postoperative drainage and hospital stay, but did not affect delayed seroma formation after axillary lymphadenectomy for breast cancer.

[Invasive pulmonary aspergillosis in neutropenic patients. Analysis of a series of 36 cases: contribution of thoracic scanners and itraconazole]. / Aspergillose pulmonaire invasive des patients neutropéniques. Analyse d'une série de 36 cas: apport du scanner thoracique et de l'itraconazole.

In neutropenic patients, one way of improving invasive pulmonary aspergillosis (IPA) prognosis is an earlier initiation of the antifungal treatment. We report our experience with 36 cases of IPA in 35 patients with haematological malignancies. When aspergillosis was diagnosed, all but 2 patients were neutropenic (PMN < 500; median duration = 20 days). The most frequent clinical signs were cough (100%), chest pain (78%) and haemoptysis (58%). Before the diagnosis of IPA, Aspergillus antibody test was positive in 60% of cases. A thoracic CT-scan was performed in 23 patients and demonstrated highly suggestive images in 22 cases (96%) with presence of CT halo sign (n = 13) or CT air-crescent sign (n = 9). Moreover, Aspergillus antigen test was positive in 13 of 28 tested patients. IPA diagnosis was determined to be definite in 23 cases and probable or possible in 13 cases. Thirty-five patients were treated: in 3 cases with amphotericin B (2 failures) and in 32 cases with itraconazole (7 failures and 25 successes or improvements). In 7 cases (including 5 emergencies) surgical resection was successfully combined with the medical treatment. In neutropenic patients with fever, we consider that thoracic CT-scan and repeated biological tests (Aspergillus antibody and antigen tests) can be used to establish the diagnosis of aspergillosis. Early treatment with itraconazole and surgical resection appears to improve prognosis.

Regional chemotherapy (with mitomycin C) and intra-operative hyperthermia for digestive cancers with peritoneal carcinomatosis.

Intraperitoneal chemo-hyperthermia with mitomycin C was used to treat 28 patients with far advanced digestive adenocarcinoma and histologically confirmed peritoneal carcinomatosis. Surgical resection of the primary tumor was possible in 17 cases. After closure of the abdominal wall, intraperitoneal chemo-hyperthermia was performed for 90 to 120 minutes under general anesthesia and 32 degrees C hypothermia, through 3 intraperitoneal drains forming a closed circuit, using 10 mg/l of mitomycin C in 6 liters of peritoneal dialysate heated to an inflow temperature of 46-49 degrees C. No mortality occurred, and there were 2 post-operative complications, with transitory biological side effects. In 9 out of 10 patients with preoperative malignant ascites, the ascites cleared after treatment. One-year survival rate was 54.2%. These encouraging preliminary results show that intraperitoneal chemohyperthermia with mitomycin C is a safe and reliable treatment for peritoneal carcinomatosis in far advanced digestive cancers.

Utility of C-reactive protein measurements for empyema diagnosis after pneumonectomy.

Serum C-reactive protein (CRP) levels were studied serially during the postoperative period in 151 consecutive patients who underwent pneumonectomy. Virtually all patients who had a simple postoperative course (115 of 120), as well as 9 patients who had a bronchial infection of the remaining lung, 3 with a pulmonary embolus, and 2 who suffered postoperative bleeding requiring reoperation, demonstrated a similar postoperative evolution in their CRP values: a rapid postoperative rise until a peak or a plateau (mean peak value, 132 +/- 25 mg/L) was reached within 3 to 6 days, followed by a progressive decline to a value of less than 75 mg/L on day 9, and less than 50 mg/L on day 12. Conversely, all 12 patients who suffered empyema postoperatively, as well as 3 patients with bacterial pneumonia, 1 patient with chylothorax, and 1 patient with inflammatory pericarditis, demonstrated either a markedly persistent elevation in their CRP values or a secondary rise in the levels which exceeded 100 mg/L. Because of the high sensitivity (100%) and specificity (91.4%) of the CRP levels in detecting postpneumonectomy empyema, we recommend the routine use of this measure. Furthermore, a low CRP value after pneumonectomy (less than 50 mg/L) may help in deciding whether to confidently discharge a patient from the hospital in the absence of empyema. The negative predictive value of this method was found to be 100%.

[Effect of fibrin glue on lymphostasis during lymph node excisions. Prospective randomized study in 40 patients]. / Efficacité d'une colle de fibrine sur la lymphostase au cours des curages ganglionnaires. Etude prospective randomisée sur 40 patients.

To evaluate the efficacy of fibrin glue for lymphostasis during axillary and inguinal lymph node removal, we conducted a prospective randomized study including 40 patients. Post-operative mortality and morbidity rates were not different throughout the 2 groups. However they were significant decreasement in drainage duration, in drainage quantity and in hospital duration in the group "with" fibrin when compared with the group "without".

Intraoperative chemohyperthermia in the management of digestive cancers. A general review of literature.

Intraoperative chemohyperthermia is a new method in the treatment of peritoneal seedings from digestive cancers, which combines surgery, intraperitoneal chemotherapy (mitomycin C and/or cisplatyl) and peritoneal hyperthermia. After a brief reminder on the general principles concerning high temperature action, a review of literature is made: 5 teams have performed this technique. We differentiate the indications, design features and results of each team. The results show a mean survival after 2 years of 35% (in peritoneal carcinomatosis) up to 78% (in gastric serosal invasion, peritoneal seeding free). The best result of the method is the drying up of cancerous ascites, allowing a more comfortable survival.

A new device for the treatment of pleural malignancies: intrapleural chemohyperthermia preliminary report.

The prognosis of malignant pleural tumors remains extremely unfavorable. The aim of this study is to evaluate the combination of intrathoracic intrapleural chemotherapy and intrapleural hyperthermia (ITCH) in these diseases. Under anesthesia, 5 men were studied. After pleurectomy for mesothelioma (3/5) or adenocarcinoma (2/5), ITCH is carried out for over 60 min, either with mitomycin C (4/5) or cisplatin (1/5). No pre- or postoperative death occurred. The maximal pleural temperature is 42.6 degrees C. The blood level of mitomycin C never reached the systemic toxic level. All the patients were discharged from the surgical ward, 3 are still alive 15 months later. Therefore, ITCH appears to be a safe and reliable therapy.

Study of the pharmacokinetics of mitomycin C in humans during intraperitoneal chemohyperthermia with special mention of the concentration in local tissues.

The aim of this work is to estimate the 24-hour distribution and elimination of mitomycin C (MMC) during and after intraperitoneal chemohyperthermia (IPCH) in 18 patients (13 gastric adenocarcinoma, 3 pancreatic adenocarcinoma, 2 malignant mesothelioma) who received 60 mg MMC during 90-120 min in 6 liters of heating solution HS; (42 degrees C) or HS flowing at 0.4 liters/min in a closed circuit. MMC assay in the serum, urine, HS and in local biopsies were performed by high performance liquid chromatography. The amount of MMC in HS decreased by 54.1 +/- 13.6% during IPCH. The maximum MMC levels in serum reached 0.4 +/- 0.18 mg/l 45 min after the start of IPCH, then rapidly decreased. Only 1.77 +/- 0.93 mg were recovered in urine in 24 h. These data are consistent with a large and rapid absorption, mostly in local tissue, demonstrated by the level in 7 post-IPCH biopsies (8.3 +/- 7.6 mg/kg).

[Cancers of the stomach and peritoneal carcinoses: will hyperthermia be a new therapeutic approach?]. / Cancers de l'estomac avec carcinoses péritonéales: l'hyperthermie sera-t-elle une nouvelle voie thérapeutique?

Overall survival rate of gastric adenocarcinoma is poor whatever the therapeutic is. Main reasons for this poor prognosis are regional failure and local recurrences. Using hyperthermia could be of interest in peritoneal carcinomatosis during gastric carcinoma: in vitro cytotoxic effect of hyperthermia is well known. Using hyperthermia in combination with intra peritoneal chemotherapy could improve local control. Preliminary results of pilot studies treating peritoneal carcinomatosis with intra peritoneal chemohyperthermia are reported.

[Intraoperative radiotherapy in gastric adenocarcinomas. Apropos of 45 cases]. / Radiothérapie per-opératoire dans les adénocarcinomes gastriques. A propos de quarante-cinq cas.

Intra-operative radiation therapy (IORT) is a multidisciplinary approach in which residual tumors or tumor beds are directly irradiated during a surgical procedure. To evaluate its efficacy we conducted since 1985 a prospective study including non metastatic gastric adenocarcinoma treated by surgery, IORT (15 Gy) and post-operative external beam radiotherapy (44 Gy). Up to now, 45 gastric adenocarcinomas (18 pNO and 27 pN1N2) have been included in the study. Mortality and morbidity rates were not different from those of surgery alone. Overall 5 year survival rate is 59.1% and pN1N2 5 years survival rate is 50.6%. These promising results are comparable with those of asian randomized studies which demonstrate the possible value of IORT in the treatment of gastric adenocarcinoma.

[Inflammatory pseudotumor of the superior vena cava: rare etiology of mediastinal tumor]. / Pseudotumeur inflammatoire de la veine cave supérieure: étiologie rare de tumeur médiastinale.

The patient reported here had no particular past history. He presented with effort dyspnea, jugular turgescence and positional vertigo suggestive of a superior vena cava obstruction syndrome. Bronchial fibroscopy was normal, but chest X-ray, CT scan and phlebography of the superior vena cava revealed an anterior mediastinal tumour compressing the superior vena cava. The tumour was excised with replacement of the vena cava by a PTFE n. 16 graft. Pathological examination showed this to be an inflammatory tumour by fibrosis with plasmocytes. This plasmocyte fibrosis is histologically benign and becomes malignant only when complications occur as a result of its spread.

[Intraperitoneal chemo-hyperthermia in the treatment of peritoneal carcinomatosis of ovarian origin. Initial cases, physiopathologic data]. / Chimio-hyperthermie intrapéritonéale dans le traitement des carcinoses péritonéales d'origine ovarienne. Premières observations, données physiopathologiques.

The authors report the first results using intraperitoneal hyperthermic chemotherapy with mitomycin C or cisplatinum with protective hypothermia up to 32 degrees C in four cases of peritoneal carcinomatosis secondary to cancer of the ovary which was resistant to conventional treatment. They were stage III and IV in the FIGO classification. There were no post-operative complications. Two patients are still alive 4 and 6 months after intraperitoneal hyper-chemotherapy. Hypothermia has its own action of destroying tumour cells and potentiates the effects of chemotherapy. We have, following the work of the japanese, devised a therapeutic regime combining chemotherapy and intraperitoneal hyperthermia.

[Intraperitoneal chemo-hyperthermia with mitomycin C. Development and in vivo study of tolerability in dogs]. / Chimio-hyperthermie intra-péritonéale (CHIP) avec mitomycine C (MMC). Mise au point et étude in vivo de la tolérance chez le chien.

Peritoneal carcinosis is relatively frequent in the clinical course of digestive cancers (48% in cases with invasion of the serous membrane). Despite various therapeutic strategies, prognosis is almost always unfavourable, survival after 6 months is extremely rare. Because of the known in vitro anti-tumour effect of hyperthermia and its chemosensitizing effect during Mitomycine C therapy, we decided to develop a technique based on these two anticancer effects: intraperitoneal hyperthermia and intraperitoneal chemotherapy. Termed CHIP, or chemo-hyperthermia intraperitoneal, this new method could be used for the treatment of peritoneal carcinosis. We report here results of the necessary animal experimentation before such a method could be proposed for human anticancer therapy.