Results of induction chemotherapy in children older than 18 months with stage-4 neuroblastoma treated with an adaptive-to-response modified N7 protocol (mN7)

Purpose. We report the response rate in children older than 18 months with stage 4 Neuroblastoma, using a modified dose-intensive, response-adaptive, induction mN7 protocol. Methods. From 2005 to 2012, 24 patients were treated with the mN7 protocol. Phase 1 included five MSKCC N7 cycles and surgery and two high-dose cyclophosphamide-topotecan (HD-CT) cycles for those who did not achieve complete remission (CR) and negative bone marrow (BM) minimal residual disease (MRD) status (CR+MRD-). Phase 2 consisted of myeloablative doses of topotecan, thiotepa and carboplatin plus hyperfractionated RT. Phase 3 included isotretinoin and 3F8 immunotherapy plus GM-CSF. BM MRD was monitored using GD2 synthase, PHOX2B and cyclin D1 mRNAs. Results. After 3 cycles, all patients showed BM complete histological clearance and 6 (25 %) were MRD-. Twenty of 21 s-look surgeries achieved macroscopic complete resection. After 5 cycles and surgery, 123I-MIBG scan was negative in 15 (62.5 %) cases, BM disease by histology was negative in 23 (96 %) and 10 (42 %) patients were MRD-. Twelve (50 %) pts were in CR, 2 in very good partial response (VGPR), 9 partial response (PR) and one had progressive disease. With 2 HD-CT extra cycles, 17 (71 %) pts achieved CR+MRD- status moving to phase 2. Overall and event-free survival at 3 years for the 17 patients who achieved CR+MRD- is 65 and 53 %, respectively, median follow-up 47 months. Seven (29 %) patients never achieved CR+MRD-. Univariate Cox regression analysis shows CR+MRD- status after mN7 induction as the only statistically significant prognostic factor to predict overall survival. Conclusions. mN7 induction regimen produced a CR+MRD- rate of 71 %. CR+MRD- status following induction was the only predictive marker of long-term survival (AU)

No disponible

Results of induction chemotherapy in children older than 18 months with stage-4 neuroblastoma treated with an adaptive-to-response modified N7 protocol (mN7).

PURPOSE: We report the response rate in children older than 18 months with stage 4 Neuroblastoma, using a modified dose-intensive, response-adaptive, induction mN7 protocol. METHODS: From 2005 to 2012, 24 patients were treated with the mN7 protocol. Phase 1 included five MSKCC N7 cycles and surgery and two high-dose cyclophosphamide-topotecan (HD-CT) cycles for those who did not achieve complete remission (CR) and negative bone marrow (BM) minimal residual disease (MRD) status (CR+MRD-). Phase 2 consisted of myeloablative doses of topotecan, thiotepa and carboplatin plus hyperfractionated RT. Phase 3 included isotretinoin and 3F8 immunotherapy plus GM-CSF. BM MRD was monitored using GD2 synthase, PHOX2B and cyclin D1 mRNAs. RESULTS: After 3 cycles, all patients showed BM complete histological clearance and 6 (25 %) were MRD-. Twenty of 21 s-look surgeries achieved macroscopic complete resection. After 5 cycles and surgery, (123)I-MIBG scan was negative in 15 (62.5 %) cases, BM disease by histology was negative in 23 (96 %) and 10 (42 %) patients were MRD-. Twelve (50 %) pts were in CR, 2 in very good partial response (VGPR), 9 partial response (PR) and one had progressive disease. With 2 HD-CT extra cycles, 17 (71 %) pts achieved CR+MRD- status moving to phase 2. Overall and event-free survival at 3 years for the 17 patients who achieved CR+MRD- is 65 and 53 %, respectively, median follow-up 47 months. Seven (29 %) patients never achieved CR+MRD-. Univariate Cox regression analysis shows CR+MRD- status after mN7 induction as the only statistically significant prognostic factor to predict overall survival. CONCLUSIONS: mN7 induction regimen produced a CR+MRD- rate of 71 %. CR+MRD- status following induction was the only predictive marker of long-term survival.

Evaluation of corneal anaesthesia after the application of topical 2 per cent lidocaine, 0.5 per cent bupivacaine and 1 per cent ropivacaine in dogs.

The objective of this study was to describe and compare the efficacy and duration of topical anaesthesia induced by 2 per cent lidocaine (L), 0.5 per cent bupivacaine (B) and 1 per cent ropivacaine (R) in the dog using 24 clinically healthy beagles with normal ocular examination. Dogs were randomly divided into three groups: 2 per cent lidocaine (n=8), 0.5 per cent bupivacaine (n=8) and 1 per cent ropivacaine (n=8). The baseline corneal touch threshold (CTT) was measured using a Cochet-Bonnet aesthesiometer. Following baseline CTT measurement, a single drop of the assigned anaesthetic was applied to each eye and the CTT was measured bilaterally within one minute after administration and every five minutes until the basal CTT value was restored. Data were analysed with non-parametric analysis of variance models and Dunnet's test for post hoc analysis. One per cent ropivacaine was the most effective drug (LCTTmax=3 cm, BCTTmax=2 cm, RCTTmax=0 cm; P<0.001), and had the shortest latency (LLatency=5 minutes, BLatency=5 minutes, RLatency=1 minute; P<0.001) and the smallest AUC (LAUC=80 cm×minute, BAUC=68.25 cm×minute, RAUC=36.88 cm×minute; P<0.001). There was no statistical difference in the duration of corneal anaesthesia between the groups (P=0.09) and all topical anaesthetics tested reduced corneal sensitivity, although 1 per cent ropivacaine had the maximal and quickest anaesthetic effect. This drug could be used for specific diagnostic procedures where quick and short, but effective, corneal anaesthesia is needed.

Shikonin promotes intestinal wound healing in vitro via induction of TGF-ß release in IEC-18 cells.

The intestinal barrier is a complex system with a dynamic structure that is designed for the maintenance of homeostasis in healthy individuals. Ulcerative colitis, one of the main manifestations of inflammatory bowel disease, is characterized by an inadequate and delayed wound healing. Shikonin, the active principle in the root of Lithospermum erythrorhizon, has demonstrated its ability to attenuate dextran sulfate sodium-induced ulcerative colitis in mice. Moreover, the root of L. erythrorhizon has been used in traditional Chinese medicine for treatment of burns, anal ulcers, hemorrhoids and skin wounds. However, the effect of shikonin on intestinal wound healing is unknown. Using an in vitro model for wound healing, we observed that shikonin enhances cell migration of intestinal epithelial cells through a mechanism that involves TGF-ß1 induction. The combination of shikonin's anti-inflammatory activity together with its wound-healing properties makes it a great potential therapeutic agent for the treatment of injury associated with intestinal inflammation.

Traditional chinese medicine remedy to jury: the pharmacological basis for the use of shikonin as an anticancer therapy.

Shikonin is the major constituent of the root of Lithospermum erythrorhizon, which has been used in traditional Chinese medicine to treat external wounds, burns, or dermatitis for centuries. Nowadays, this root is commonly used as an herbal medicine against cancer. Studies carried out over the past 30 years have demonstrated that many of the effects historically associated with the use of this root have a scientific basis, with shikonin and its derivatives being responsible for its pharmacological properties. These include both anti-inflammatory and anticancer effects. While previous summaries have focused on the pharmacokinetics and toxicity of shikonin, the aim of this review is to report on the most current findings with regard to shikonin's antitumor activity by summarizing and comparing the various studies published in the last ten years and discussing the pharmacological aspects that make shikonin a promising anticancer agent.

Anti-inflammatory agents from plants: progress and potential.

The identification of substances that can promote the resolution of inflammation in a way that is homeostatic, modulatory, efficient, and well-tolerated by the body is of fundamental importance. Traditional medicines have long provided front-line pharmacotherapy for many millions of people worldwide. Medicinal extracts are a rich source of therapeutic leads for the pharmaceutical industry. The use of medicinal plant therapies to treat chronic illness, including rheumatoid arthritis (RA) and inflammatory bowel disease (IBD), is thus widespread and on the rise.The aim of this review is to present recent progress in clinical anti-inflammatory studies of plant extracts and compound leads such as green tea polyphenols, curcumin, resveratrol, boswellic acid, and cucurbitacins, among others, against chronic inflammatory diseases, mainly RA and IBD. In this context, the present paper also highlights the most promising experimental data on those plant extracts and pure compounds active in animal models of the aforementioned diseases.

Secondary plant substances in various extracts of the leaves, fruits, stem and bark of Caraipa densifolia Mart.

Thirty secondary plant substances were detected in various extracts of the leaves, fruits, stem and bark of Caraipadensifolia Mart. Phenolic compounds were preliminarily identified and quantitated by HPLC-ESI-MS and the structures of the compounds, purified by semi-preparative HPLC, were further characterized by nano-ESI-MS-MS. The presence of gallic acid, 3,4-dihydroxybenzoic acid, neochlorogenic acid, chlorogenic acid, methyl gallate, p-coumaric acid quinate, epicatechin, procaynidin dimer B(2), procyanidin trimer C(1), syringic acid, 1,2,3,6-tetragallate glucoside, 1,3,4,6-tetragallate glucoside, corilagin, ellagic acid, methyl ellagic acid rhamnoside, quercetin-3-O-rhamnoside, two apigenin-C-glycosides (vitexin and isovitexin) and two luteolin-C-glycosides (orientin and isoorientin) are reported in this species for the first time. In addition, the previously reported following terpenoids, lupeol, lupenone, betulinic acid, betulin, friedelin and a previously non-characterized terpenoid in this species, friedelinol were identified and quantitated by GC-MS. A previously identified sterol was beta-sitosterol along with stigmasterol in this species for the first time. The vitamins alpha-tocopherol and gamma-tocopherol were also identified in extracts of the leaves of Caraipa species for the first time. The data shows that the botanical parts of C. densifolia Mart. has a much richer spectrum of secondary plant substances than previously reported.

Headspace solid-phase microextraction of oil matrices heated at high temperature and phthalate esters determination by gas chromatography multistage mass spectrometry.

A solvent-free analytical approach based on headspace solid-phase microextraction (SPME) of oil matrices heated at high temperatures coupled to gas chromatography with mass spectrometry detector (GC-ion trap) has been developed for the determination of phthalic acid esters (PAEs) in oil matrices without sample manipulation. For this study, three fibers, i.e., 85 microm-polyacrylate (PA), 50/30 microm-divinylbenzene-carboxen-polydimethylsiloxane (DVB/CAR/PDMS) and 100 microm-polydimethylsiloxane (PDMS) were tested. Variables affecting the SPME headspace composition such as incubation sample temperature, sample incubation time and fiber exposition time were optimized. The optimal values found were 250 degrees C for sample incubation temperature and 30 min for incubation and extraction time. PA fiber was not suitable for the lightest polar phthalates which showed poor extraction and repeatability values. PDMS fiber had very poor response for some of the heavier and non-polar phthalates, whereas DVB/CAR/PDMS fiber showed the best response and repeatability values for the majority of the phthalates studied. The main benefit of the analytical method proposed is the absence of sample manipulation and hence avoidance of possible contamination coming from glassware, environment, solvents and samples.

Increased transcutaneous oxygen tension in the skin dorsum over the foot in patients with diabetic foot disease in response to the topical use of an emulsion of hyperoxygenated Fatty acids.

The aim of this study was to examine changes in the skin over the feet of patients with diabetic foot syndrome after local application of a product containing hyperoxygenated fatty acids (HOFAs) by measuring transcutaneous oxygen. In 64 patients, transcutaneous oxygen pressure (TcPo(2)) was measured on days 0, 7, 30, 60, and 90 of the study. Foot skin dryness, shedding, and skin color were also assessed using a clinical score. The patients were grouped on the basis of initial levels of transcutaneous oxygen; group 1 comprised patients with TcPo( 2) >30 mm Hg and group 2 comprised patients with TcPo(2) <30 mm Hg on the skin over the dorsum of the feet. Increases in local oxygenation values were observed at a local level in group 2 patients after 30 days of treatment. Skin trophism showed clinical improvement in all patients and these observations may be attributed to improved local microcirculation.

Inhibition of transcription factors by plant-derived compounds and their implications in inflammation and cancer.

Inflammation is a general term used to describe various pathological processes with diverse causes that can include infection, trauma, or an autoimmune response. Due to its many causes, the inflammatory response involves multiple and varied mediators, including vasoactive amines, free radicals, and both lipidic and peptidic mediators. Medicinal plants and the compounds derived from them are a good source of new and specific inhibitors of the inflammatory process. The past decade has witnessed many important discoveries in this field, with new findings challenging the more traditional views of pharmacologists. Various studies, for example, have demonstrated the positive effects of plant-derived phenolics, which act as anti-oxidants, free radical scavengers, and inhibitors of nitric-oxide synthase, cyclooxygenase, and lipoxygenase. The anti-inflammatory activity of chalcones has been correlated with the induction of heme oxygenase-1 while phlorotannins have been found to inhibit matrix metalloproteinase, which is implicated in arthritis, chronic inflammation, and wrinkle formation. Sesquiterpene lactones have been studied as inhibitors of NF-kappaB activity and the relationship between their chemical structure and pharmacological activity has been clearly established. Recently, cucurbitacins have been described as inhibitors of JAK -STAT and NF-AT functions related to inflammation; they were also found to induce apoptosis of cells involved in the inflammatory response. This review focuses mainly on the effects of natural products on transcription factors, which are the most promising targets for designing new active drugs against inflammation and cancer.

In vitro and in vivo effects of Ranunculus peltatus subsp. baudotii methanol extract on models of eicosanoid production and contact dermatitis.

Ranunculus (Crowfoot) species are numerous and they are all reputed to be counter-irritants and are used in several topical conditions. In order to study the pharmacological mechanisms of action underlying this popular use, a methanol extract of Ranunculus peltatus was tested in vitro in various assays involving eicosanoid and human elastase release by intact cells as well as in vivo, with models of delayed-type hypersensitivity (DTH) contact dermatitis. The extract proved to be a selective inhibitor of the cyclooxygenase-1 pathway, producing the total inhibition of 12-(S)-HHTrE release at 200 microg/mL, while leaving both 5-lipoxygenase and 12-lipoxygenase activities unaffected at the same dose. The n-hexane, chloroform and ethyl acetate fractions of the crude methanol extract inhibited LTB(4) release by intact rat peritoneal neutrophils, but more polar fractions were inactive and did not increase the 5-LOX activity as seen previously for extracts of other Ranunculus species. In the in vivo models, the methanol extract reduced the dinitrofluorobenzene (DNFB)-induced oedema by 40%, but failed to inhibit the oedema brought on by oxazolone. The results agree with the age-old assertion that Water Crowfoot species can be used as a topical antiinflammatory remedy without the prominent irritant action that accompanies the application of non-aquatic Ranunculus species.

Demethylnobiletin inhibits delayed-type hypersensitivity reactions, human lymphocyte proliferation and cytokine production.

BACKGROUND AND PURPOSE: Our aim was to examine the effect of demethylnobiletin on various experimental models of delayed-type hypersensitivity (DTH) reactions and to determine its influence on the mediators and enzymes involved in these reactions. EXPERIMENTAL APPROACH: DTH was induced in mice by oxazolone, dinitrofluorobenzene (DNFB) and sheep red blood cells (SRBC). The effect of demethylnobiletin on the ensuing DTH was studied, especially in relation to oedema formation, cell infiltration and tissue damage. Its activity on different mediators implicated in DTH reactions was also determined and its effect on nitric oxide synthase (NOS)-2 analysed. Finally, its influence on T lymphocyte proliferation, apoptosis and caspase 3 activity was tested. KEY RESULTS: DTH reactions were all reduced by demethylnobiletin. The experimental results suggest that the compound may act by reducing cell infiltration and by suppressing mediators such as interleukin-2 (IC50=1.63 microM), interleukin-4 (IC50=2.76 microM), tumour necrosis factor-alpha (IC50=0.66 microM), interferon-gamma (IC50=1.35 microM), and interleukin-1 beta (46% at 2.5 microM) and by concomitantly increasing the production of the anti-inflammatory cytokine, interleukin-10. In addition, while demethylnobiletin affected nitric oxide production, it did not modify NOS-2 expression. Finally, demethylnobiletin inhibited proliferation of T cells and induced their apoptosis. CONCLUSIONS AND IMPLICATIONS: Demethylnobiletin decreased DTH reactions induced by various agents. This finding, along with the fact that the compound has a low toxicity and exhibits several other interesting properties, could pave the way for other structurally related citroflavonoids to be used as pharmacological agents in complementary therapies.

Solid-phase extraction gas chromatography-ion trap-mass spectrometry qualitative method for evaluation of phenolic compounds in virgin olive oil and structural confirmation of oleuropein and ligstroside aglycons and their oxidation products.

Phenolic compounds in Spanish virgin olive oil were analyzed by GC-MS after an SPE diol cartridge extraction and clean-up procedure. Posterior derivatization to trimethylsilyl (TMS) ethers using a mixture of hexamethyldisilazane:dimethylclorosilane (HMDS:DMCS) in pyridine (3:1:9) was performed. Several compounds were detected and 21 of them were identified. Free phenols such as hydroxytyrosol, tyrosol, tyrosyl and hydroxytyrosyl acetate, and aldehydic and dialdehydic forms of elenolic acid linked to tyrosol and hidroxytyrosol were the most abundant compounds. Likewise, oxidation products coming from the aldehydic and dialdehydic forms of elenolic acid, and of ligstroside and oleuropein aglycons, were detected, and their structure confirmed by other mass spectrometry technique, i.e., HPLC-APCI-MS. Individual oxidation products were isolated from an oxidized sample by preparative HPLC, converted to TMS ethers and re-analyzed by GC-MS. When necessary and for identification purposes, selective ion monitoring, namely, GC-MS-SIM, was employed. This is the first time that structures of oxidized forms are determined by GC-MS.

Medicinal plants and antimicrobial activity.

In the present paper, we analyze the past, present and future of medicinal plants, both as potential antimicrobial crude drugs as well as a source for natural compounds that act as new anti-infection agents. In the past few decades, the search for new anti-infection agents has occupied many research groups in the field of ethnopharmacology. When we reviewed the number of articles published on the antimicrobial activity of medicinal plants in PubMed during the period between 1966 and 1994, we found 115; however, in the following decade between 1995 and 2004, this number more than doubled to 307. In the studies themselves one finds a wide range of criteria. Many focus on determining the antimicrobial activity of plant extracts found in folk medicine, essential oils or isolated compounds such as alkaloids, flavonoids, sesquiterpene lactones, diterpenes, triterpenes or naphtoquinones, among others. Some of these compounds were isolated or obtained by bio-guided isolation after previously detecting antimicrobial activity on the part of the plant. A second block of studies focuses on the natural flora of a specific region or country; the third relevant group of papers is made up of specific studies of the activity of a plant or principle against a concrete pathological microorganism. Some general considerations must be established for the study of the antimicrobial activity of plant extracts, essential oils and the compounds isolated from them. Of utmost relevance is the definition of common parameters, such as plant material, techniques employed, growth medium and microorganisms tested.

Inhibition of Trypanosoma cruzi by plant extracts used in Chinese medicine.

In this work, we assessed the effect of extracts obtained from 17 plants used in traditional Chinese medicine. These extracts were tested in vitro with the epimastigote form of Trypanosoma cruzi, clone Bra C(15) C(2), at 27 degrees C in F-29 medium at a concentration of 100 microg/ml in axenic cultures. Allopurinol was used as reference drug. Seven plant extracts showed inhibitory activities lower than 25%. Pueraria lobata, Mahonia beaei, Dictamus dasycarpus, Kochia scoparia, Sophora flavescens and Ligustrum lucidum showed effects with inhibition values between 25% and 60%, whereas Lithospermum erythrorhizon, Saussurea lappa, Melia toosendan and Cinnamomum cassia showed the greatest inhibitory activity of 100%. The IC(50) of these extracts were: 0.4, 2.4, 1.8 and 3.9 microg/ml, respectively. The MTT assay was made and did not show cytotoxic activity. These results allowed us to suggest that L. erythrorhizon, S. lappa, M. toosendan and C. cassia could be a source of new compounds against T. cruzi.

Dual inhibition of cyclooxygenase-1 and 5-lipoxygenase by aerial part of Bupleurum fruticescens methanol extract.

The effect of the methanol extract from aerial parts of Bupleurum fruticescens on the release of eicosanoids and hydrolytic enzymes was determined on in vitro cell systems. The extract had a significant effect on 5-lipoxygenase (5-LOX) activity, inhibiting both LTB4 and 5(S)-HETE production with IC50 values of 112 microg/ml and 95 microg/ml, respectively. At concentrations of 200 microg/ml, the extract also inhibited cyclooxygenase-1 (90%) and elastase activities (54%). The 12-LOX activity in intact platelets was not affected; a fact, which suggests that phospholipase A2 (PLA2) activity, is not modified by the extract.

[Ultrasound-guided hydrostatic reduction for the treatment of idiopathic intestinal invagination]. / Reducción hidrostática guiada por ecografía para el tratamiento de la invaginación intestinal idiopática.

The more extended conservative treatment in intussusception consists of the accomplishment of fluoroscopy guided neumoenema. The development of the echography has allowed to introduce this as alternative in diagnosis and treatment of intussusception. Our objective is to analyze the therapeutic value of echography during resolution of intussusception with saline solution enema in pediatric patients. Intestinal intussusception was diagnosed in 183 patients by echography during a period of 7 years. Patients are classified in three groups according to treatment by means of opaque enema, radioscopy guided neumoenema or echography guided hydrostatic reduction enema. The results of the three series are analyzed: rate of failure of the procedure, the recurrences or the presence of complications. We exclude n = 18 patients that were operated on directly after the diagnosis. The reduction rate was 81.25% (9/16) with opaque enema; 94.6% (103/110) with neumoenema and 96.4% (27/28) with echography guided saline enema. Complications appeared in n = 2 cases of intestinal perforation, 1 with barium and 1 with air 2. Recurrences were n = 3, n = 2 with neumoenema and n = 1 with echography guided enema. Echography guide saline enema has a high security and effectiveness (96.4%), similar to the other methods, avoiding the exposition to ionic radiation. The technique is safe and it does not increase the rate of recurrences.

Influence of traditional Chinese anti-inflammatory medicinal plants on leukocyte and platelet functions.

The enzymes 5-lipoxygenase and elastase are therapeutic targets in dermatological disorders such as psoriasis. Fifteen extracts from traditional Chinese medicinal plants used to treat topical inflammations were screened for their inhibitory effect on lipoxygenase, cyclooxygenase and elastase activity in intact leukocytes and platelets. Astragalus membranaceus, Forsythia suspensa and Poria cocos inhibited 5-lipoxygenase, with IC50 values of 141, 80 and 141 microg mL(-1), respectively. The latter two species, along with Angelica dahurica and Angelica pubescens, also inhibited elastase (IC50 values of 80, 123, 68 and 93 microg mL(-1), respectively), while A. pubescens, Atractylodes macrocephala, Lentinus edodes, Rehmannia glutinosa and Paeonia lactiflora selectively inhibited 12-(S)-HHTrE production, a valid marker of cyclooxygenase activity. The inhibition of phospholipase A(2) activity by P. cocos is discussed. Dehydrotumulosic and pachymic acids, which have been isolated from P. cocos, were shown to inhibit leukotriene B(4) release. The results indicate that both P. cocos and F. suspensa are potentially valuable species in the management of skin pathologies involving chronic inflammation.

Ecotoxicological evaluation of carbamazepine using six different model systems with eighteen endpoints.

The occurrence of pharmaceutically active compounds in the aquatic environment has been recognized as one of the emerging issues in environmental chemistry. However, the ecotoxicological effects of pharmaceuticals have still not been researched adequately. Carbamazepine, an anticonvulsant commonly present in surface and groundwater, was studied, using six ecotoxicological model systems with eighteen endpoints evaluated at different exposure time periods. The battery included the immobilization of Daphnia magna, bioluminescence inhibition in the bacterium Vibrio fischeri, growth inhibition of the alga Chlorella vulgaris, and micronuclei induction and root growth inhibition in the plant Allium cepa. Cell morphology, neutral red uptake, total protein content, MTS metabolization, lactate dehydrogenase leakage and activity and glucose-6-phosphate dehydrogenase activity were studied in the salmonid fish cell line RTG-2. The total protein content, LDH activity, neutral red uptake and MTT metabolization in Vero monkey kidney cells were also investigated. The most sensitive system to carbamazepine was the Vero cell line, followed by Chlorella vulgaris, Vibrio fischeri, Daphnia magna, Allium cepa, and RTG-2 cells. EC50 values from 19 microM in Vero cells at 72 h to more than 1200 microM in other systems, were obtained. Comparing the concentrations in water and the toxicity quantified in our assay systems, carbamazepine is not expected to produce acute toxic effects in the aquatic biota under these circumstances, but chronic and synergistic effects with other chemicals cannot be excluded.