Wnt addiction of genetically defined cancers reversed by PORCN inhibition.

Enhanced sensitivity to Wnts is an emerging hallmark of a subset of cancers, defined in part by mutations regulating the abundance of their receptors. Whether these mutations identify a clinical opportunity is an important question. Inhibition of Wnt secretion by blocking an essential post-translational modification, palmitoleation, provides a useful therapeutic intervention. We developed a novel potent, orally available PORCN inhibitor, ETC-1922159 (henceforth called ETC-159) that blocks the secretion and activity of all Wnts. ETC-159 is remarkably effective in treating RSPO-translocation bearing colorectal cancer (CRC) patient-derived xenografts. This is the first example of effective targeted therapy for this subset of CRC. Consistent with a central role of Wnt signaling in regulation of gene expression, inhibition of PORCN in RSPO3-translocated cancers causes a marked remodeling of the transcriptome, with loss of cell cycle, stem cell and proliferation genes, and an increase in differentiation markers. Inhibition of Wnt signaling by PORCN inhibition holds promise as differentiation therapy in genetically defined human cancers.

Incidence and predictors of radial artery occlusion after transradial coronary angioplasty: Doppler-guided follow-up study.

OBJECTIVES: To evaluate the incidence and predictors of radial artery occlusion (RAO) after transradial coronary angioplasty (TRA). BACKGROUND: RAO can occur after TRA but has not been well studied by serial vascular Doppler examination. METHODS: A total of 198 patients undergoing TRA were included. Radial pulse and Doppler examination of the radial artery were performed 1 day before, 1 day after, and 3 months after the procedure. RAO was defined as an absence of antegrade flow on Doppler studies. Logistic regression analysis was done to evaluate the predictors of RAO. RESULTS: The mean radial arterial diameter was 2.8 ± 0.4 mm. On the day after TRA, radial artery Doppler examination revealed RAO in 30 patients (15.2%). Radial pulse was still palpable in 30.0% of these patients. All of them were asymptomatic. At 3-month follow-up, no new RAO was noted. Interestingly, the radial artery had spontaneously recanalized in 8 patients (26.7%) with RAO. Patients with persistent RAO remained asymptomatic. On univariate analysis, female sex, diabetes, lower body mass index, radial artery diameter ≤2.5 mm, lower peak systolic velocity, and radial artery to sheath ratio <1 were predictors of RAO. Interestingly, procedural characteristics and duration of the procedure were not identified as predictors of RAO. On multivariate analysis, radial artery diameter ≤2.5 mm and preprocedural peak systolic velocity emerged as independent predictors for RAO. CONCLUSION: Asymptomatic RAO occurs in about 15% of patients after TRA. Spontaneous recanalization occurs in about one-fourth of these patients. Preprocedure radial artery inner diameter ≤2.5 mm and peak systolic velocity are the independent predictors of RAO.

An overview on traditional uses and pharmacological profile of Acorus calamus Linn. (Sweet flag) and other Acorus species.

Acorus calamus (Sweet flag) has a long history of use and has numerous traditional and ethnomedicinal applications. Since ancient times, it has been used in various systems of medicines such as Ayurveda, Unani, Siddha, Chinese medicine, etc. for the treatment of various aliments like nervous disorders, appetite loss, bronchitis, chest pain, colic, cramps, diarrhea, digestive disorders, flatulence, gas, indigestion, rheumatism, sedative, cough, fever, bronchitis, inflammation, depression, tumors, hemorrhoids, skin diseases, numbness, general debility and vascular disorders. Various therapeutic potentials of this plant have been attributed to its rhizome. A number of active constituents from leaves, rhizomes and essential oils of A. calamus have been isolated and characterized. Of the constituents, alpha and beta-asarone are the predominant bioactive components. Various pharmacological activities of A. calamus rhizome such as sedative, CNS depressant, anticonvulsant, antispasmodic, cardiovascular, hypolipidemic, immunosuppressive, anti-inflammatory, cryoprotective, antioxidant, antidiarrheal, antimicrobial, anticancer and antidiabetic has been reported. Genotoxicity and mutagenecity of beta and alpha-asarone is reported, which limits their use at high dosage. Though A. calamus has been used since ancient times, many of its uses are yet to be scientifically validated. In the present review an attempt has been made to explore traditional uses and pharmacological properties of A. calamus.

Third trimester amniocentesis for diagnosis of inherited bleeding disorders prior to delivery.

X-linked and autosomally inherited bleeding disorders confer a risk of foetal intracranial haemorrhage during delivery. Conventional prenatal diagnosis involving chorionic villus sampling or early amniocentesis is primarily aimed at offering the choice of pregnancy termination. Currently, non-invasive procedures, involving analysis of free foetal DNA in the maternal circulation, are restricted to gender determination, and are of limited value in women at risk of carrying a foetus with a bleeding disorder. These limitations, together with the rising proportion of women shown to be carrying an affected foetus, who decide to continue the pregnancy, have led to the development of prenatal mutation identification via late amniocentesis after 34 weeks of gestation, with the sole aim of directing delivery management. Although this approach has been documented in some cases of potential foetal anomaly, there are no previous reports of its use in women with heritable bleeding disorders. We report a single-centre experience of this technique in managing nine such deliveries. Of these, three showed an affected foetus, five showed an unaffected foetus and in one case no result could be obtained. In the three affected cases and the one with the inconclusive result restrictive birth plans were implemented, whereas the five unaffected cases underwent routine obstetric management; with one delivery necessitating interventions which would have been contraindicated if foetal status had not been determined. Late amniocentesis is a safe technique for guiding delivery management in women with bleeding disorders where the mutation is known.

SB1518, a novel macrocyclic pyrimidine-based JAK2 inhibitor for the treatment of myeloid and lymphoid malignancies.

SB1518 is an innovative pyrimidine-based macrocycle that shows a unique kinase profile with selective inhibition of Janus Kinase-2 (JAK2; IC50=23 and 19 nM for JAK2(WT) and JAK2(V617F), respectively) within the JAK family (IC50=1280, 520 and 50 nM for JAK1, JK3 and TYK2, respectively) and fms-like tyrosine kinase-3 (FLT3; IC50=22 nM). SB1518 shows potent effects on cellular JAK/STAT pathways, inhibiting tyrosine phosphorylation on JAK2 (Y221) and downstream STATs. As a consequence SB1518 has potent anti-proliferative effects on myeloid and lymphoid cell lines driven by mutant or wild-type JAK2 or FLT3, resulting from cell cycle arrest and induction of apoptosis. SB1518 has favorable pharmacokinetic properties after oral dosing in mice, is well tolerated and significantly reduces splenomegaly and hepatomegaly in a JAK2(V617F)-driven disease model. SB1518 dose-dependently inhibits intra-tumor JAK2/STAT5 signaling, leading to tumor growth inhibition in a subcutaneous model generated with SET-2 cells derived from a JAK2(V617F) patient with megakaryoblastic leukemia. Moreover, SB1518 is active against primary erythroid progenitor cells sampled from patients with myeloproliferative disease. In summary, SB1518 has a unique profile and is efficacious and well tolerated in JAK2-dependent models. These favorable properties are now being confirmed in clinical studies in patients with myelofibrosis and lymphoma.

Pacritinib (SB1518), a JAK2/FLT3 inhibitor for the treatment of acute myeloid leukemia.

FMS-like tyrosine kinase 3 (FLT3) is the most commonly mutated gene found in acute myeloid leukemia (AML) patients and its activating mutations have been proven to be a negative prognostic marker for clinical outcome. Pacritinib (SB1518) is a tyrosine kinase inhibitor (TKI) with equipotent activity against FLT3 (IC(50)=22 n) and Janus kinase 2 (JAK2, IC(50)=23 n). Pacritinib inhibits FLT3 phosphorylation and downstream STAT, MAPK and PI3 K signaling in FLT3-internal-tandem duplication (ITD), FLT3-wt cells and primary AML blast cells. Oral administration of pacritinib in murine models of FLT3-ITD-driven AML led to significant inhibition of primary tumor growth and lung metastasis. Upregulation of JAK2 in FLT3-TKI-resistant AML cells was identified as a potential mechanism of resistance to selective FLT3 inhibition. This resistance could be overcome by the combined FLT3 and JAK2 activities of pacritinib in this cellular model. Our findings provide a rationale for the clinical evaluation of pacritinib in AML including patients resistant to FLT3-TKI therapy.

Consensus protocol for the use of recombinant activated factor VII [eptacog alfa (activated); NovoSeven] in elective orthopaedic surgery in haemophilic patients with inhibitors.

Patients with haemophilia complicated by inhibitors have a significant burden of joint disease, which is associated with a negative impact on their quality of life. Successful elective orthopaedic surgery can result in decreased bleed frequency into a new joint, less time spent in hospital, increased mobility and improved well being. This paper describes a new protocol for use of recombinant activated factor VII (rFVIIa) in elective orthopaedic surgery, based on a review of published data as well as the personal experience of a group of expert physicians. The protocol offers guidance on the planning of the surgery and preoperative testing as well as the bolus schedule for rFVIIa and advice on the concomitant use of antifibrinolytic agents and fibrin sealants. A total of 10 operations involving 13 procedures in eight patients in five comprehensive care centres have been undertaken until now using the protocol, which employs an initial bolus dose of rFVIIa in the range of 120-180 microg kg(-1) to cover surgery. The clinical experience reported here encompasses all cases of elective orthopaedic surgery using rFVIIa as initial treatment carried out in the UK and Republic of Ireland over the last 2 years. In all cases, there was good control of haemostasis during surgery and the final outcome was rated as 'excellent' or 'extremely satisfactory' by the reporting clinicians. Although the initial cost of product to cover surgery such as arthroplasty is high, it needs to be borne in mind that this may be offset in subsequent years by savings resulting from avoidance of bleeding episodes in the affected joint.

A randomized phase III comparative trial of immediate consolidation with high-dose chemotherapy and autologous peripheral blood progenitor cell support compared to observation with delayed consolidation in women with metastatic breast cancer and only bone metastases following intensive induction chemotherapy.

The prognosis for patients with metastatic breast cancer remains poor. Metastatic breast cancer confined to the bones may have a better prognosis, especially hormone receptor-positive disease. We performed a prospective, randomized clinical trial to compare immediate consolidation with high-dose chemotherapy and hematopoietic support versus observation with high-dose consolidation at the time of disease progression in women with metastatic breast cancer and only bone metastases. The patients received chemotherapy with doxorubicin, 5-fluorouracil and methotrexate before randomization. In all, 85 patients were enrolled and 69 were randomized. The median follow-up is 8.1 years from randomization. The median event-free survival (EFS) for the immediate transplant arm is 12 months and for the observation arm is 4.3 months (P<0.0001). The median overall survival for the immediate transplant arm is 2.97 years and for the observation arm 1.81 years, a difference that is not statistically significant. Immediate high-dose chemotherapy and radiation therapy as consolidation offers a clinically and statistically significant improvement in EFS compared with radiation therapy alone following induction chemotherapy for women with metastatic breast cancer confined to the bones.

Intravenous immunoglobulin given to lymphoma patients with recurrent haemolytic transfusion reactions after transfusion of compatible blood.

Accelerated destruction of red cells after transfusion of compatible blood has been reported in both sickle cell disease (SCD) and non-SCD patients. We report three patients with lymphoma, all of whom had recurrent haemolytic transfusion reactions after receiving compatible red cell units. The direct antiglobulin test (DAT) was negative and there were no detectable red cell alloantibodies in either pre-transfusion or post-transfusion samples. As there was no evidence of red cell antibody-mediated haemolysis and response to oral steroids, a trial of intravenous immunoglobulin (IVIg) was given. Immediate cessation of haemolysis with sustained haemoglobin level was achieved in all cases. The response to IVIg in these cases suggests that IVIg should be tried when recurrent non-antibody mediated haemolytic transfusion reactions occur in patients with a lymphoid malignancy.

Hydration heat capacity of nucleic acid constituents determined from the random network model.

The heat capacities of hydration (dCp) of the five nucleic acid bases A, G, C, T, and U, the sugars ribose and deoxyribose, and the phosphate backbone were determined using Monte Carlo simulations and the random network model. Solute-induced changes in the mean length and root mean square angle of hydrogen bonds between hydration shell waters were used to compute dCp for these solutes. For all solutes the dCp is significantly more positive than predicted from accessible surface area (ASA) models of heat capacity. In ASA models, nitrogen, oxygen, and phosphorus atoms are considered as uniformly polar, therefore making a negative contribution to dCp. However, the simulations show that many of these polar atoms are hydrated by water whose hydrogen bonds are less distorted than in bulk, leading to a positive dCp. This is in contrast to the effect of polar groups seen previously in small molecules and amino acids, which increase the water H-bond distortion, giving negative dCp contributions. Our results imply that dCp accompanying DNA dehydration in DNA-ligand and DNA-protein binding reactions may be significantly more negative than previously believed and that dehydration is a significant contributor to the large decrease in heat capacity seen in experiments.

Curcuma longa activates NF-kappaB and promotes adhesion of neutrophils to human umbilical vein endothelial cells.

Upregulation of expression of cell adhesion molecules, such as ICAM-1, VCAM-1 and E-selectin, is important for immune surveillance. Extravasation and migration of body's effector cells to the site of immune activation is controlled by the expression of cell adhesion molecules on endothelial cells. We demonstrate here that an aqueous extract prepared from Curcuma longa (ClAqE), a dietary component, promotes the adhesion of peripheral neutrophils to human umbilical vein endothelial cells. To delineate the mechanism of increased adhesion, we investigated the possibility that ClAqE induces the expression of ICAM-1 and E-selectin on endothelial cells. ClAqE increases the steady state transcript levels of ICAM-1, VCAM-1, and E-selectin as determined by RT-PCR. We also show that ClAqE activates nuclear transcription factor NF-kappaB, a major transcription factor involved in the transcription of genes encoding ICAM-1, VCAM-1 and E-selectin. These results have implications for the usage of aqueous preparation of C. longa for upregulation of cell adhesion molecule expression and/or NF-kappaB.

2'-hydroxychalcone inhibits nuclear factor-kappaB and blocks tumor necrosis factor-alpha- and lipopolysaccharide-induced adhesion of neutrophils to human umbilical vein endothelial cells.

Inhibition of expression of cell adhesion molecules (CAM), including intercellular CAM-1 (ICAM-1), vascular CAM-1 (VCAM-1), and E-selectin, has been shown to be important in controlling various inflammatory diseases. The cell adhesion proteins are induced by various inflammatory cytokines, such as tumor necrosis factor-alpha, interleukin-1, and bacterial lipopolysaccharide. The induction process primarily takes place at the level of transcription, where nuclear factor-kappaB (NF-kappaB) plays a major role. We demonstrate here that 2'-hydroxychalcone inhibits the adhesion of peripheral neutrophils to the endothelial cell monolayers by inhibiting the expression of ICAM-1, VCAM-1, and E-selectin in a concentration-dependent manner. The inhibition by 2'-hydroxychalcone is reversible. 2'-hydroxychalcone inhibits the induction of steady-state transcript levels of ICAM-1, VCAM-1, and E-selectin by tumor necrosis factor-alpha as determined by reverse transcription-polymerase chain reaction, and therefore it may interfere with the transcription of their genes. Because NF-kappaB is a major transcription factor involved in CAM expression, we studied its status in the 2'-hydroxychalcone treated cells. We demonstrate that 2'-hydroxychalcone inhibits the activation of NF-kappaB. These results have implications for using NF-kappaB inhibitors for the treatment of various inflammatory diseases.

Changes in water structure induced by a hydrophobic solute probed by simulation of the water hydrogen bond angle and radial distribution functions.

In order to better characterize changes in water structure induced by a hydrophobic solute the oxygen-oxygen and hydrogen-hydrogen radial distribution functions (goo(r), ghh(r)) and the hydrogen bond angle distribution function p(theta) for water molecules in the first hydration shell of the tetramethyl ammonium (TMA) cation were computed using Monte Carlo simulations. goo(r) and ghh(r) were corrected for the effect of solute volume exclusion on the local solvent density so that intrinsic structural changes independent of local solvent density variations could be detected. Comparison of ghh(r) of TMA's first hydration shell water with ghh(r) for bulk water shows subtle but clear evidence of structure formation induced by the ion. These changes in ghh(r) are very similar to those seen experimentally for larger tetra-alkyl ammonium ions in previous neutron diffraction experiments. Larger changes in p(theta) in the first hydration shell of TMA were seen. Comparison of changes in p(theta) with changes in goo(r) and ghh(r) show that the angle distribution function provides the most sensitive way to analyze water structure changes associated with hydrophobic solvation.

Detection of lymphoma in bone marrow by whole-body positron emission tomography.

Positron emission tomography (PET) is a whole-body imaging technique using 18 fluorine-fluorodeoxyglucose (FDG), whose uptake is increased in tumor cells. Published studies have shown PET to be an effective method of staging lymphoma and to be more sensitive than CT at detecting extranodal disease. The purpose of this study was to determine whether the increased marrow uptake of FDG observed in some lymphoma patients during routine staging PET scans represented marrow involvement by disease. PET scans of 50 patients with Hodgkin's (12) and non-Hodgkin's (38) lymphoma were analyzed by three independent observers and the marrow graded as normal or abnormal using a visual grading system. Unilateral iliac crest marrow aspirates and biopsies were performed on all patients. The PET scan and marrow histology agreed in 39 patients (78%), being concordant positive in 13 and concordant negative in 26 patients. In 8 patients the PET scan showed increased FDG uptake but staging biopsy was negative; in 4 of these 8 patients the PET scan showed a normal marrow background with focal FDG "hot spots" distant from the site biopsied. In 3 patients the marrow biopsy specimen was positive but the PET scan normal; 2 of these 3 patients had non-Hodgkin's lymphoma whose malignant cells did not take up FDG at lymph node or marrow disease sites. Therefore, there were only 5 patients (10%) in whom there was a difference between the PET scan and biopsy result which could not be fully explained. Visual interpretation of marrow FDG uptake during whole-body staging PET scans can correctly assess marrow disease status in a high proportion of lymphoma patients. PET has the potential to reduce the need for staging marrow biopsy.