Refinement of lentiviral vector for improved RNA processing and reduced rates of self inactivation repair

Background Lentiviral gene therapy vectors are now finding clinical application. In order to fully exploit their potential it is important that vectors are made as efficient and as safe as possible. Accordingly, we have modified a previously reported vector to improve RNA processing, minimise Human Immunodeficiency Virus Type-1 (HIV-1) sequence content and reduce repair of the self inactivating (SIN) deletion. Results HIV-1 sequence in the vector was reduced by substituting the polyadenylation signal with a heterologous signal. Mutation of splice donor sites was undertaken to prevent the majority of splicing within the vector genomic RNA. In addition, a number of other sequences within the vector were deleted. The combination of these modifications was able to significantly reduce the rates of both vector mobilisation and repair of the self inactivating deletion after two rounds of marker rescue. Conclusion RNA processing can be improved by mutation of the major and minor HIV-1 splice donor sites in the vector. In addition the rate of vector mobilisation and repair of SIN vectors can be successfully reduced by careful vector design that reduces homology between the 5' and 3' long terminal repeats (LTRs) to a minimum.Rachel M Koldej and Donald S Anso

Efficacy of cognitive behaviour therapy versus anxiety management for body dysmorphic disorder: a randomised controlled trial

Background: The evidence base for the effectiveness of cognitive behaviour therapy (CBT) for treating Body Dysmorphic Disorder (BDD) is weak. Aims: To determine if CBT is more effective than anxiety management (AM) in an out-patient setting. Method: A single blind, stratified parallel-group randomized controlled trial. The primary endpoint was at 12 weeks, and the Yale Brown Obsessive Compulsive Scale for BDD (BDD-YBOCS) was the primary outcome measure. Secondary measures for BDD included the Brown Assessment of Beliefs (BABS), the Appearance Anxiety Inventory (AAI) and the Body Image Quality of Life Inventory (BIQLI). The outcome measures were collected at baseline and week 12. The CBT group, unlike the AM group, had 4 further weekly sessions that were analysed for their added value. Both groups then completed measures at their 1-month follow-up. Forty-six participants, with DSM-IV diagnosis of BDD including those with a delusional BDD were randomly allocated to either CBT or AM. Results: At 12 weeks, CBT was found to be significantly superior to AM on the BDD-YBOCS ( = -7.19, S.E. () = 2.61, p < .01, C.I. = -12.31, -2.07, d 0.99) as well as the secondary outcome measures of the BABS, AAI and BIQL. Further benefits occurred by Week 16 within the CBT group. There were no differences in outcome for those with delusional BDD or depression. Conclusions: CBT is an effective intervention for people with BDD even with delusional beliefs or depression and is more effective than anxiety management over 12 weeks

Development of a liquid chromatography tandem mass spectrometry method for the simultaneous measurement of voriconazole, posaconazole and itraconazole

Background Azole-based antifungals are the first-line therapy for some of the most common mycoses and are now also being used prophylactically to protect immunocompromised patients. However, due to variability in both their metabolism and bioavailability, therapeutic drug monitoring is essential to avoid toxicity but still gain maximum efficacy. Methods Following protein precipitation of serum with acetonitrile, 20  µL of extract was injected onto a 2.1 × 50 mm Waters Atlantis dC18 3  µm column. Detection was via a Waters Quattro Premier XE tandem mass spectrometer operating in ESI-positive mode. Multiple reaction monitoring (MRM) detected two product ions for each compound and one for each isotopically labelled internal standard. Ion suppression, linearity, stability, matrix effects, recovery, imprecision, lower limits of measuring interval and detection were all assessed. Results Optimal chromatographic separation was achieved using gradient elution over 8 minutes. Voriconazole, posaconazole and itraconazole eluted at 1.71, 2.73 and 3.41 min, respectively. The lower limits of measuring interval for all three compounds was 0.1 mg/L. The assay was linear to 10 mg/L for voriconazole (R2 = 0.995) and 5 mg/L for posaconazole (R2 = 0.990) and itraconazole (R2 = 0.991). The assay was both highly accurate and precise with % bias of voriconazole, posaconazole and itraconazole, respectively, when compared with previous NEQAS samples. The intra-assay precision (CV%) was 1.6%, 2.5% and 1.9% for voriconazole, posaconazole and itraconazole, respectively, across the linear range. Conclusion A simple and robust method has been validated for azole antifungal therapeutic drug monitoring. This new assay will result in a greatly improved sample turnaround time and will therefore vastly increase the clinical utility of azole antifungal drug monitoring. </jats:sec

The Effect of Luteolin on Human Glioblastoma

Glioblastoma multiforme (GBM) is widely recognized as the most common and lethal of the malignant gliomas. Few effective therapeutic treatments are available as five-year survival rates of diagnosed individuals are less than five percent. Luteolin, a common flavonoid found in a variety of fruits and vegetables, has demonstrated significant promise in combating cancers of the breast, colon, liver, lung, and bone. In this study, we investigated the effects of luteolin on glioblastoma multiforme cell lines U-251, U-87, and U-1242. Cell viability was assessed using cell count with trypan blue exclusion and MTT assays. Results revealed that luteolin reduces GBM cell viability and cell proliferation in a time and concentration-dependent manner. Western Blot analysis indicated that luteolin decreased AKT, ERK, and MAPK phosphorylation following treatment with EGF. Additionally, luteolin promoted apoptosis in GBM cells by inducing PARP and caspase-3 cleavage, and decreasing levels of the anti-apoptotic protein BCL-XL. Our results indicate that luteolin exhibits a biological effect and may be used as a therapeutic agent for glioblastoma multiforme

A Critical Analysis of the Theoretical Basis of Ultrasonic Scattering Measurements

There are three elements involved in the backscattering from inhomogeneous media; the scattering properties of a single particle or scattering element, the scattering associated with a group of such particles and the relationship of the scattered wave to the measured signal. Ideally it should be possible to obtain information about the material microstructure from ultrasonic backscattering measurements. However, a number of assumptions and approximations must be made before the problem becomes tractable, and it is the purpose of the present investigation to compare the various approaches available in the literature in an attempt to quantify the errors involved with some of these approximations

Assisted Self-Assembly to Target Heterometallic Mn-Nd and Mn-Sm SMMs: Synthesis and Magnetic Characterisation of [Mn7_{7}Ln3_{3}(O)4_{4}(OH)4_{4}(mdea)3_{3}(piv)9_{9}(NO3_{3})3_{3}] (Ln=Nd, Sm, Eu, Gd)**

Assisted self-assembly by using a preformed [Mn$_{6}$O$_{2}$(piv)$_{10}$(4-Me-py)$_{2}$(pivH)$_{2}$] (1) (pivH=pivalic acid) cluster leads to 3 different outcomes of the reaction with Ln(NO$_{3}$)$_{3}$ ⋅ xH$_{2}$O (Ln=Pr−Ho and Y): Mn$_{7}$Ln$_{3}$ and in some cases the inverse butterfly Mn$_{2}$Ln$_{2}$ for Ln=Pr−Eu, Mn$_{5}$Ln$_{4}$ for Ln=Tb-Ho and in the special case of Gd all three possibilities can form. Whilst the magnetic properties for the Mn$_{5}$Ln$_{4}$ systems show the expected known SMM properties, for Mn$_{7}$Ln$_{3}$ two new examples of Nd- and Sm-containing SMMs have been discovered. in an assisted self-assembly approach starting from the [Mn$_{6}$O$_{2}$(piv)$_{10}$(4-Me-py)$_{2}$(pivH)$_{2}$] cluster a family of Mn−Ln compounds (Ln=Pr−Yb) was synthesised. The reaction of [Mn$_{6}$O$_{2}$(piv)$_{10}$(4-Me-py)$_{2}$(pivH)$_{2}$] (1) with N-methyldiethanolamine (mdeaH$_{2}$) and Ln(NO$_{3}$)$_{3}$ ⋅ 6H$_{2}$O in MeCN generally yields two main structure types: for Ln=Tb−Yb a previously reported Mn$_{5}$Ln$_{4}$ motif is obtained, whereas for Ln=Pr−Eu a series of Mn$_{7}$Ln$_{3}$ clusters is obtained. Within this series the Gd$^{III}$ analogue represents a special case because it shows both structural types as well as a third Mn$_{2}$Ln$_{2}$ inverse butterfly motif. Variation in reaction conditions allows access to different structure types across the whole series. This prompts further studies into the reaction mechanism of this cluster assisted self-assembly approach. For the Mn$_{7}$Ln$_{3}$ analogues reported here variable-temperature magnetic susceptibility measurements suggest that antiferromagnetic interactions between the spin carriers are dominant. Compounds incorporating Ln=Nd$^{III}$(2), Sm$^{III}$(3) and Gd$^{III}$ (5) display SMM behaviour. The slow relaxation of the magnetisation for these compounds was confirmed by ac measurements above 1.8 K

Terminal Ligand and Packing Effects on Slow Relaxation in an Isostructural Set of [Dy(H2_{2}dapp)X2_{2}]+ Single Molecule Magnets**

Three new dysprosium complexes with a pentadentate ligand occupying five equatorial sites differ only in the nature of the axial ligands. These help tune the relaxation properties as judged by an analysis of the AC susceptibility data. More in depth analysis by using two recently suggested fitting equations lead to similar outcomes for all three systems. As a further contribution to the relaxation pathway involving the phonon bath it is concluded that a short nitrate-nitrate interaction between molecules helps dampen the spin phonon coupling. We report three structurally related single ion Dy compounds using the pentadentate ligand 2,6-bis((E)-1-(2-(pyridin-2-yl)-hydrazineylidene)ethyl)pyridine (H$_{2}$dapp) [Dy(H$_{2}$dapp)(NO$_{3}$)$_{2}$]NO$_{3}$ (1), [Dy(H$_{2}$dapp)(OAc)$_{2}$]Cl (2) and [Dy(H$_{2}$dapp)(NO$_{3}$)$_{2}$]Cl$_{0.92}$(NO$_{3}$)$_{0.08}$ (3). The (H$_{2}$dapp) occupies a helical twisted pentagonal equatorial arrangement with two anionic ligands in the axial positions. Further influence on the electronic and magnetic structure is provided by a closely associated counterion interacting with the central N−H group of the (H$_{2}$dapp). The slow relaxation of the magnetisation shows that the anionic acetates give the greatest slowing down of the magnetisation reversal. Further influence on the relaxation properties of compounds1 and 2 is the presence of short nitrate-nitrate intermolecular ligand contact opening further lattice relaxation pathways

A microscopic theory of gauge mediation

We construct models of indirect gauge mediation where the dynamics responsible for breaking supersymmetry simultaneously generates a weakly coupled subsector of messengers. This provides a microscopic realization of messenger gauge mediation where the messenger and hidden sector fields are unified into a single sector. The UV theory is SQCD with massless and massive quarks plus singlets, and at low energies it flows to a weakly coupled quiver gauge theory. One node provides the primary source of supersymmetry breaking, which is then transmitted to the node giving rise to the messenger fields. These models break R-symmetry spontaneously, produce realistic gaugino and sfermion masses, and give a heavy gravitino.Comment: 24 pages, 2 figures, accepted to JHEP for publicatio

Syntheses, Crystal Structure, Electrocatalytic, and Magnetic Properties of the Monolanthanide-Containing Germanotungstates [Ln(H2_{2}O)n_{n}GeW11_{11}O39_{39}]5−^{5-} (Ln = Dy, Er, n = 4,3)

Two monolanthanide-containing polyanions based on monolacunary Keggin germanotungstates [Ln(H2O)nGeW11O39]5− (Ln = Dy, Er, n = 4,3) have been synthesized in simple one-pot synthetic procedure and compositionally characterized in solid state by single-crystal X-ray diffraction, powder X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, and elemental analysis. Electronic absorption and emission spectra of the title compounds in solution were also studied. The [DyIII(H2O)4GeW11O39]5− Keggin POM exhibits a slow relaxation of magnetization. The cyclic voltammetry measurements and mass spectrometry were carried out to check the stability of the compounds in solution. Both polyanions prove efficient in the electrocatalytic reduction of nitrite. To our knowledge, this observation establishes the first example of electrocatalysis of nitrite reduction by all inorganic monolanthanidecontaining germanotungstates family

Stabilization of inorganic mixed waste to pass the TCLP and STLC tests using clay and pH-insensitive additives

Stabilization traps toxic contaminants (usually both chemically and physically) in a matrix so that they do not leach into the environment. Typical contaminants are metals (mostly transition metals) that exhibit the characteristic of toxicity. The stabilization process routinely uses pozzolanic materials. Portland cement, fly ash-lime mixes, gypsum cements, and clays are some of the most common materials. In many instances, materials that can pass the Toxicity Characteristic Leaching Procedure (TCLP-the federal leach test) or the Soluble Threshold Leachate Concentration (STLC-the California leach test) must have high concentrations of lime or other caustic material because of the low pH of the leaching media. Both leaching media, California`s and EPA`s, have a pH of 5.0. California uses citric acid and sodium citrate while EPA uses acetic acid and sodium acetate. These media can form ligands that provide excellent metal leaching. Because of the aggressive nature of the leaching medium, stabilized wastes in many cases will not pass the leaching tests. At the Lawrence Livermore National Laboratory, additives such as dithiocarbamates and thiocarbonates, which are pH-insensitive and provide resistance to ligand formation, are used in the waste stabilization process. Attapulgite, montmorillonite, and sepiolite clays are used because they are forgiving (recipe can be adjusted before the matrix hardens). The most frequently used stabilization process consists of a customized recipe involving waste sludge, clay and dithiocarbamate salt, mixed with a double planetary mixer into a pasty consistency. TCLP and STLC data on this waste matrix have shown that the process matrix meets land disposal requirements