A retrospective case series of 103 consecutive patients with leptomeningeal metastasis and breast cancer.

Approximately 2-5 % of patients with breast cancer (BC) develop leptomeningeal metastasis (LM). 103 consecutive patients with BC were diagnosed with LM and initially treated with intra-CSF liposomal cytarabine from 2007 to 2011 at a single institution. Correlations were determined with respect to patient characteristics and BC subtype with regard to overall survival (OS). At LM diagnosis, 61 % of patients had a 0-2 performance status (PS), the remaining 39 % were severely neurologically impaired. Regardless of PS, all patients received intra-cerebrospinal fluid (CSF) liposomal cytarabine as first-line treatment. Systemic treatment and radiotherapy were also given in 58 and 17 % of patients respectively as clinically appropriate. Second- (intra-CSF thiotepa) and third-line (intra-CSF methotrexate) treatment was administered in 24 and 6 patients respectively. Median OS was 3.8 months (range 1 day-2.8 years). In multivariate analysis, an initial combined treatment, a second-line treatment with intra-CSF thiotepa, an initial clinical response, and a non-'ER/PR/HER2 negative' BC were significantly associated with a better OS. Median OS in this heterogeneous retrospective case series was similar to that of previously observed BC patients treated with intra-CSF methotrexate suggesting intra-CSF liposomal cytarabine is a reasonable first choice therapy of BC-related LM.

Megestrol acetate versus metronomic cyclophosphamide in patients having exhausted all effective therapies under standard care.

BACKGROUND: To evaluate the antitumour activity and safety of metronomic cyclophosphamide vs megestrol acetate in progressive and advanced cancer patients having exhausted all effective therapies under standard care. METHODS: Patients were randomly assigned to receive orally metronomic cyclophosphamide (50 mg b.i.d) or megestrol acetate (160 mg only daily) until intolerance or progression (RECIST 1.0). The primary efficacy end point was a 2-month progression-free rate (PFR(2m)). According to Optimal Simon's design and the following assumptions, namely, P0=5%, P1=20%, alpha=beta=10%, the treatment is considered as effective if atleast 5 out of 44 patients achieved PFR(2m). RESULTS: Between September 2006 and January 2009, 88 patients were enrolled. Two patients experienced grade 3-4 toxicities in each arm (4%). One toxic death occurred in the megestrol acetate arm as a consequence of thrombosis. The metronomic cyclophosphamide arm reached the predefined level of efficacy with a PFR(2m) rate of 9 out of 44 and a PFR(4m) rate of 5 out of 44. The MA arm failed to achieve the level of efficacy with a PFR(2m) of 4 out of 44 and a PFR(4m) of 1 out of 44. The median overall survival was 195 and 144 days in the metronomic cyclophosphamide arm and megestrol acetate arm, respectively. CONCLUSION: Metronomic cyclophosphamide is well tolerated and provides stable disease in such vulnerable and poor-prognosis cancer patients. This regimen warrants further evaluations.

Additional direct medical costs associated with nosocomial infections after head and neck cancer surgery: a hospital-perspective analysis.

The clinical impact of surgical site infections (SSI) and postoperative pneumonia (PP) after head and neck cancer surgery has been assessed in the past, but little is known about their economic impact. The present study was designed to evaluate costs related to SSI and PP after head and neck cancer surgery with opening of mucosa. The incidence of SSI and PP was measured in a prospective cohort of 261 patients who had undergone head and neck cancer surgery. The additional direct medical costs related to these infections from the hospital perspective were determined based on postoperative length of stay. The mean direct hospital costs for patients with and without SSI or PP were compared. Of the 261 patients, 81 (31%), 21 (8%) and 13 (5%) developed SSI, PP or both, respectively. The additional lengths of stay attributable to SSI, PP or both were 16, 17 and 31 days, respectively, and additional direct medical costs related to these conditions were 17,000, 19,000 and 35,000 Euros. Nosocomial infections after head and neck cancer surgery significantly increase patients' length of stay and therefore generate additional direct medical costs. These results support the application of preventive interventions to reduce nosocomial infections in this setting.

[Analysis of the factors influencing the internal reporting of nosocomial infections. A review of 108 notifications]. / Analyse des facteurs influençant la démarche de signalement interne d'infection nosocomiale. A propos de 108 signalements.

INTRODUCTION: Since July 26, 2001, the external reporting to the regional office of health and social affairs (Direction départementale des affaires sanitaires et sociales--Ddass) and the coordination centre (Comité de lutte contre les infections nosocomiales--Cclin) for the fight against nosocomial infections (NI) is mandatory. However, the modalities of internal reporting to the Clin are unknown. METHOD: We performed a retrospective analysis of 108 cases of NI reported over 23 months in 4 medical-surgical departments (MSD) with 14 to 35 NI reported/MSD. The distribution of the bacteria responsible was compared with that of the local epidemiological state (chi2 test). A correlation analysis was performed between the number of NI reported in each MSD and the structural characteristics and activity index of these MSD (Spearmann's correlation test). RESULTS: The NI were predominantly infections related to a catheter (43), lower respiratory tract (25) and infection of the site of surgery (19). Ninety were documented biologically, among which 10 implied multi-resistant bacteria. Ninety-four NI were associated with the prescription of an antibiotic. Compared with the local epidemiological state, the NI reported generally implied multi-resistant bacteria (p=0.009). The other microbiological data had little implication. In each of the MSD, the number of cases reported was independent of: the global activity, the number of interventions, the mean duration of hospitalisation, the number of beds, the number of clinicians, the number of new patients managed and the chemotherapy outpatient activity. Conversely, there was a strong correlation between the global consumption of antibiotics (r=0.78), and the number of the Clin members in each MSD DMC (r=0.82). CONCLUSION: In each MSD, the internal reporting of NI relies on the discovery of multi-resistant bacteria, but above all on the implication of those involved in the fight against nosocomial infections.

Effects of Rosa canina fruit extract on neutrophil respiratory burst.

Respiratory burst leads polymorphonuclear neutrophils (PMN) to produce reactive oxygen species (ROS) such as superoxide anions (O(2)(o-)), hypochlorous acid (HOCl) and hydrogen peroxide (H(2)O(2)) which may possess deleterious effects for the organism. Rosa canina fruits are well known to contain a large amount of vitamin C which is antioxidant. This study was focused on the polyphenolics contained in rose hips to evaluate their antioxidative properties. We prepared a rose hip extract deprived of vitamin C. The extract contained mainly phenolics such as proanthocyanidins and flavonoids. We investigated its effects directly against (O(2)(o-)), HOCl and H(2)O(2) and investigated its effects on isolated PMN. For that, in vitro inflammatory conditions were reproduced by stimulating PMN with stimuli having different transductional pathways, in order to determine a possible mechanism of action. The results showed that the extract can inhibit ROS tested in acellular and cellular systems. The IC(50) obtained were 5.73 mg/L, 1.33 mg/L and 2.34 mg/L respectively for (O(2)(o-)), HOCl and H(2)O(2) in acellular experiments. For cellular experiments, the IC(50) were quite similar. Thus, the extract did not present an effect on PMN metabolism. Therefore, the antioxidative effects of Rosa canina are due not only to vitamin C but also to polyphenolics.

Investigation of the inhibitory effects of HA-1077 and Y-32885 on the translocation of PKCbetaI, PKCbetaII and PKCzeta in human neutrophils.

To transmit the information inside the cell, one possibility is the action of an enzyme called kinase that phosphorylates other proteins. To study these enzymes, chemical compound synthesis was needed to know the function and the mechanism of activation. The major difficulty is creating a specific molecule for one kinase. In this study, we test the action of Rho-kinase inhibitors (HA-1077 and Y-32885) on protein kinase C (PKC) in the respiratory burst in the human polymorphonuclear neutrophils. We have shown that these compounds could inhibit the anion superoxide production. To prove their action on PKC, we have shown a decrease of binding of a specific ligand (phorbol-12,13-dibutyrate) with each inhibitor. During its activation, PKC was translocated from the cytoplasm to the plasmic membrane. We have also shown an inhibition of this translocation, proving an inhibition of PKC by HA-1077 and Y-32885.

Investigation of the inhibitory effects of chelerythrine chloride on the translocation of the protein kinase C betaI, betaII, zeta in human neutrophils.

The protein kinase C (PKC) is a serine/threonine kinase, consisting of different isoforms, implicated in numerous processes of signal transduction. To understand this enzyme well, different pharmacological tools were developed. To activate PKC specifically, phorbol esters were previously used but recent research has shown that these compounds are able to stimulate other proteins. Our model is the respiratory burst in the polymorphonuclear neutrophils. A decrease in the inflammatory process was measured using chelerythrine chloride. Action on PKC was proved by a binding study and by showing the absence of translocation of this enzyme from the cytoplasm to the plasmic membrane during stimulation.

A simple method for isolating human and rabbit polymorphonuclear neutrophils (PMNs).

We report the successful application to human venous blood of a novel method developed to purify rabbit polymorphonuclear neutrophils (PMNs) from whole blood. Human PMNs were separated from whole blood after sedimentation with dextran and histopaque density gradient centrifugation. 3.92 +/- 0.26 x 10(6) PMNs per ml of blood was harvested. The purity of the preparation was 92.00 +/- 1.10%. The PMNs isolated were capable of generating a high amount of reactive oxygen species (ROS) and elastase after stimulation with phorbol 12-myristate 13-acetate (PMA): 13.43 +/- 0.3 microM of O2-, 9.62 +/- 0.15 microM of H2O2 and 5.48 +/- 0.01 microM of elastase. This method gives equivalent yield and viability when applied to isolating human or rabbit PMNs, in comparison with standard methods used to isolate human PMNs. Our method could be usefully exploited for comparative studies of rabbit and human PMNs with a cellular model of inflammatory oxidative stress in which the monitoring parameters are ROS and elastase. Thus, the results of animal (rabbit) studies can be extended to human diseases.

Reduced ammonium chloride haemolysis time enhances the number of isolated functional rabbit polymorphonuclear neutrophils.

In the present study we compared seven different methods for isolating rabbit polymorphonuclear neutrophils (PMNs) with a view to assessing viability, lymphocyte contamination and isolation yield. The two methods offering the best isolation yield and functional PMNs were retained. Leukocyte-containing plasma fraction was obtained after erythrocyte sedimentation with dextran. First, PMNs were isolated from this fraction, using hypotonic ammonium chloride haemolysis followed by Histopaque density gradient centrifugation (Method-A). Second, PMNs were obtained from leukocyte-containing plasma after centrifugation on two Percoll layers (Method-B). These processes resulted in a high cellular yield: 2.66x10(6)+/-0.22 PMNs per ml of blood (Method-A) and 1.87x10(6)+/-0.37 PMNs per ml of blood (Method-B). In both cases the PMNs isolated were of high purity and viability. In comparison, when using the standard techniques for rabbit - consisting of ammonium chloride haemolysis taking at least four times as long--fewer PMNs were isolated. The PMNs isolated by Method-A and -B were able to generate a high amount of reactive oxygen species (ROS) after stimulation with phorbol 12-myristate 13-acetate (PMA). These methods to separate PMNs are recommended for in vitro studies.

Phenolic compounds and antioxidant activities of buckwheat (Fagopyrum esculentum Moench) hulls and flour.

The interest of polyphenolics as therapeutic agents against diseases involving radical damage is growing. The phenolic contents of the hulls and flour from the seeds of Fagopyrum esculentum (French variety 'La Harpe') (total phenols, flavonoids, total flavanols, oligomeric proanthocyanidins) are compared with the antioxidative effects of the extracts against reactive oxygen species: hydrogen peroxide, hypochlorous acid, superoxide anion. The higher efficiency of the flour extract can be related to its higher flavanolic content rather than to flavonoids which are predominant in the hull extract.

High-performance liquid chromatographic determination of naltrexone in plasma of hemodialysis patients.

A simple, sensitive, selective and reliable reversed-phase high-performance liquid chromatographic (HPLC) method with UV detection is described for the determination of naltrexone in plasma samples. Naltrexone and the internal standard, naloxone, were isolated from plasma either with a liquid-liquid extraction method using ethyl acetate or with a solid-phase extraction method using Sep-Pack C18 cartridge before chromatography. The extracts were dried under a stream of nitrogen and the samples were reconstituted in the mobile phase, then 20 microL were injected on a Waters Symmetry C18 column (5 microm particle size, 4.6 x 150 mm). The mobile phase consisted of 0.06% triethylamine (pH 2.8)-acetonitrile (92:8, v/v) pumped at 1 mL/min. The peak-area ratio versus plasma concentration was linear over the range of 10-500 ng/mL and the detection limit was less than 8 ng/mL. Quantification was by ultra-violet detection at 204 nm. The present method was applied to the determination of the plasma concentration of naltrexone in dialyzed patients. Patients (n = 8) with severe generalized pruritus received 50 mg of naltrexone orally per day for 2 weeks. The variability in the therapeutic response in treated patients required plasma concentration investigations of this opioid antagonist.

Neurosedative and antioxidant activities of phenylpropanoids from ballota nigra.

Ballota nigra is a European plant known for its neurosedative properties. In this study, the ability of five phenylpropanoids (verbascoside, forsythoside B, arenarioside, ballotetroside, and caffeoyl malic acid) isolated from a hydroalcoholic extract, to bind to benzodiazepine, dopaminergic, and morphinic receptors was investigated. To carry out these studies, affinity tests with rat striata, entire brains and receptor rich preparations were employed. In addition, the phenolic aspect of these five phenylpropanoid esters led to investigate antioxidant activities using cell-free experiments and cellular experiments including isolated polymorphonuclear neutrophils (PMN). Effects of phenylpropanoid esters against reactive oxygen species as superoxide anion, peroxide hydrogen, hypochlorous acid and hydroxyl radical were tested. These molecules are liberated by PMN during inflammatory disorders, so that reproduction of this process in vitro stimulating PMN by chemical stimulants was undertaken. Results show that four of the five compounds are able to bind to the studied receptors. Inhibitory concentrations at 50% were determined and vary from 0.4 to 4.7 mg/ml. This may be in relation with the Ballota nigra known neurosedative activities. Results concerning antioxidant investigations evidence an ability to scavenge reactive oxygen species. Inhibitory concentrations at 50% obtained are comparable to those of known antioxidant drugs (mesna or N-acetyl cysteine). Moreover, the use of different stimuli having various pathways of action on PMN oxidative metabolism permits to establish that each phenylpropanoid ester has its own particular way of action by using proteine kinase C or phospholipase C pathways.

Effects of two antihypertensive agents, labetalol and metoprolol, on the production of reactive oxygen species by normal polymorphonuclear leukocytes in vitro.

OBJECTIVE: Increased lipid peroxidation is a putative causal factor for preeclampsia. Because oxygen free radicals are involved in inducing the lipid oxidation chain reaction, we evaluated two beta adrenoreceptor blockers, labetalol and metoprolol, currently used for treating hypertension with regard to their ability to inhibit the formation of reactive oxygen species during respiratory burst of human normal polymorphonuclear leukocytes in vitro. METHODS: We determined whether labetalol or metoprolol have antioxidative activity in a model of polymorphonuclear leukocytes stimulated in vitro with phorbol-12-myristate-13-acetate (PMA) and N-formyl-methionin-leucin-phenylalanin (fMLP). We also studied the scavenging properties of these two drugs using acellular systems. RESULTS: Labetalol inhibits O2-. production by neutrophils activated by fMLP (IC50 = 17.5 mg/L) and weakly by PMA (43.6% inhibition at 100 mg/L). It also possesses a significant activity on OH. production (IC50 = 65 mg/L) that may depend in part on its ability to interfere with iron in the Fenton reaction. The same assays performed with metoprolol did not show any inhibitory effect on O2.- generation. It decreased weak OH. production by neutrophils, as a result of cellular and scavenging effects. CONCLUSION: Labetalol shows important antioxidative properties in vitro with regard to normal leukocyte oxidative metabolism.

Antioxidant properties of albumin: effect on oxidative metabolism of human neutrophil granulocytes.

The present study aims at investigating the effect of bovine serum albumin (BSA) on the trial of oxidative-stress. The antioxidant effects of BSA were determined by human neutrophil granulocytes oxygen free radicals and their by-products (O2-, H2O2, HOCl) productions. BSA interacts with those reactive oxygen species (ROS) in a dose-dependent manner. The 50% inhibitory concentration (IC50) of BSA estimated, after phorbol-12-myristate-13-acetate (PMA) stimulation were: 33.5 mg/ml for O2-, 6.5 mg/ml for H2O2, and 6.85 mg/ml for HOCl. When neutrophils were washed after pre-incubation with BSA, there was no significant decrease of ROS after stimulation of PMA (maximal: 15 +/- 1.2%). In the free cell experiments, IC50 for H2O2 and HOCl were 7.86 mg/ml and 0.67 mg/ml, respectively. The mechanism at which BSA acts may result from a simple chemical interaction with ROS rather than an intracellular mechanism by intervention in PMA oxidative metabolism. These antioxidant activities confer to BSA properties, which might be used to prevent damage inflicted by these ROS during inflammatory disorders.

Serum opioid activity after physical exercise in rats.

In order to study the effect of exercise on the total serum opioid activity, female rats were trained for 3 weeks on a motor-driven treadmill and the experiment was ended by a final strenuous run until exhaustion. The serum samples were taken immediately after the final run and were analyzed by radioreceptor assay. Despite considerable interindividual variations, serum opioid activity, expressed in met-enkephalin equivalents (ME eq +/- S.D.), was significantly higher in the exercising group (74.5+/-50.5 pmol ME eq/ml) than in the control group (35.7+/-20.2 pmol ME eq/ml). Because of the much lower molar levels of beta endorphin and met-enkephalin, this result suggests that many other opioid peptides might be involved in that increase.

Implementing a multiple-isolator unit for centralized preparation of cytotoxic drugs in a cancer center pharmacy.

Due to numerous reasons: assuring safety (technicians, patients, nurses, environment), preventing medication errors, cost, maintaining pharmaceutical quality, rules and regulations, it was decided to create a dedicated room within the pharmacy for the preparation of intravenous cytotoxic drugs. After a preliminary study, the following choices were made: isolator unit instead of a vertical laminar air-flow hood, rigid surface instead of flexible film, a multiple-unit structure (one half suit unit for storage and one isolator unit for passthrough, two preparation units, each with four double-gloving portals) instead of a single-unit structure. After the equipment was installed, the physical and microbiological processes were validated and a medical/pharmaceutical catalog of protocols was created. Then the technicians were trained and the standard operating procedures were written. Updated every six months, they describe the general organization, gaseous sterilization of the isolators, the drugs and the medical devices, prescription analysis and circuit, preparation of nominative forms and labels, double checking the preparations delivery, cleaning and maintenance, documentation and reports, waste disposal, safety and protection and instructions for emergency. The pharmacists, pharmacy staff, physicians and nurses were all included in a work group responsible for the isolator unit project. The unit was opened in January 1997.

Evaluation of the effects of Gd complexes used as magnetic resonance imaging contrast agents, on superoxide dismutase: comparison of two methods.

Investigations are currently being made into the safety of gadolinium complex contrast agents used in Magnetic Resonance Imaging. Their hyperosmolality or potential Gd3+ release is evoked as a cause of various anaphylactoid reactions to be observed in humans after intravenous injection. An estimation has already been made of their effects on the liberation of reactive oxygen intermediates by neutrophils. The purpose of this study was to find a suitable method to measure SOD activity in the presence of these hyperosmolar solutions, and to evaluate their action on this activity. Two techniques were compared to measure this activity. Results and statistical analysis showed that pyrogallol autoxidation was greatly affected by solution osmolalities, whereas ferricytochrome C reduction was not. Gadopentetate dimeglumine and gadoterate meglumine seemed to activate Cu, Zn SOD in vitro, but did not exhibit any SOD-like activity. Gadodiamide did not interfere with this system of detoxication.

Pharmacokinetics of midazolam: comparison of sublingual and intravenous routes in rabbit.

In France, the legal routes used to administer midazolam to a patient are intravenously and intramuscularly. For anaesthetists, these routes are not well adapted to paediatric use; they lead to pain at injection site and stress on children. The sublingual route should be a good compromise between stress and quick efficiency. We have developed a sublingual tablet of midazolam. The aim of the present investigation is to compare the pharmacokinetic parameters of midazolam tablets administered by the sublingual and intravenous routes in 6 rabbits to determine the bioequivalence between these routes. We have estimated the 1-hydroxy-midazolam serum level by difference between RRA and HPLC values. By the sublingual route, midazolam absorption is substantial and fast. The statistical analysis, on data obtained with HPLC dosage, shows no significant difference between pharmacokinetic parameter values calculated after intravenous and sublingual administration (0.5 mg). The absolute bioavailability was close to 100%. With RRA dosage, however, AUCs were greater than those obtained by HPLC dosage (174%). 1-hydroxy-midazolam seems to have a great importance in BZD activity. To estimate the bioequivalence between intravenous and sublingual midazolam administration, it is necessary to take into account the active metabolites.

Time and concentration dependent influence of dirithromycin on neutrophils oxidative burst.

Dirithromycin is a 14-membered macrolide antibiotic, well known to yield high intragranulocytic levels after several hour exposure. We chose therefore to investigate oxidative metabolism after prolonged incubation periods with neutrophils. Neutrophil generation of reactive oxygen species, represented by superoxide anion, was assessed after fMLP or Staphylococcus aureus-induced activation of the respiratory burst. Cellular uptake of the drug was assessed concurrently, in order to attempt a correlation with time-dependent modifications of the cellular oxidative status. For 1 hour exposure time, a pro-oxidant effect was reported for lower concentrations, achievable during therapeutic administration, whereas the highest ones promoted a potent anti-oxidant effect. After prolonged incubation times, the anti-oxidant effect alone was reported, with time-dependent modifications of IC50 values. These values could be correlated with intracellular accumulation of the drug. The anti-inflammatory activity reported here for high dirithromycin concentrations, could be nevertheless clinically relevant, since dirithromycin cellular uptake extends beyond 4 hours.

Effects of PVC bags sterilization process on the 5-fluorouracil stability.

The stability and compatibility of 5-fluorouracil (5-FU) in undiluted or diluted admixtures stored in beta-radiation sterilized portable poly(vinyl chloride) (PVC) infusion bags were investigated. Admixtures containing 5-FU 50 mg ml(-1) not diluted or 25 mg ml(-1) diluted in 0.9% sodium chloride injection were placed in 100 or 250 ml empty PVC reservoirs sterilized initially by beta-irradiation. They were protected from light and placed at 37 degrees C. Two ml quantities were withdrawn immediately after preparation and after storage for 1, 2, 3, 4, 5, 6, 7 and 14 days. For each condition, samples from each admixture were tested for drug concentration by stability-indicating high-performance liquid chromatography. The admixtures were also monitored for precipitation, color change and pH. Evaporative water loss from the containers was also measured. 5-FU was compatible with PVC containers in all tested conditions for 14 days. No loss of drug and no color change were detected throughout the storage period. pH values were stable and neither precipitation nor loss of water through the reservoirs was observed when drug 50 or 25 mg ml(-1) (diluted using 0.9% sodium chloride) was stored in 100 ml capacity polyvinyl PVC bags. However, when stored in 250 ml capacity PVC bags, the 5-FU solution showed precipitation after 13 and 14 days of storage, but no drug loss was detected due to a substantial loss of water. The precipitation of the drug was due to the decrease of pH induced by the dehydrochlorination of PVC during beta-irradiation leading to the formation of hydrochloric acid in solution. Differences observed between 100 and 250 ml capacity bags can be explained by the greater area of PVC present in 250 ml reservoirs, and consequently more HCl formed. Finally, more plasticizer, di-(2-ethylhexyl) phthalate (DEHP), was then detected in drug solutions stored in 250 ml PVC bags. So, we recommend the use of 100 ml bags to store 5-FU at longer storage times and higher temperatures.