LIPOSOMAL TECHNOLOGIES TO IMPROVE GENE DELIVERY

Lipid based nanoparticles (LBNs) are used in myriad applications in medicine from small molecule drug delivery to mRNA vaccines. A major contributing factor to the development of the field has been the ongoing development of novel compounds that retain the functionality of natural lipids but expand upon them through inclusion of functional moieties that can be applied to specific scientific and biomedical questions. In the body of this dissertation, an extensive overview of LBNs is provided, focusing primarily on their use in immune modulation. The research presented herein begins with the synthesis of a novel class of lipids based on the triazine (TZ) cyanuric chloride. Twelve compounds were synthesized and assessed for their biophysical behavior and ability to form LBNs. Of the 12 compounds, 10 were able to form nanoparticles and these were assessed for in vitro toxicity. The toxicity of the nanoparticles differs based on the nanoparticle charge and approximate that observed for similarly charged compounds. The cationic TZ lipids were then tested in vitro for their ability to deliver plasmid DNA into cells where they showed improved efficacy compared with the cationic lipid DOTMA, and similar toxicity. Finally, TZ lipids were used to lipidate peptides in a liposomal peptide vaccine where they induced similar anti-peptide titers as a CHEMS conjugate. Following these experiments, the in vivo toxicity and potential for plasmid delivery was evaluated for the cationic TZ lipids. TZ lipids led to toxicity similar to other cationic lipids. Of note, the PEG length in the nanoparticles was studied for its effect on transfection efficiency as was the effect of the helper lipid in the formulation. These experiments showed improved transfection efficiency with DOPE and with shorter length PEG chains on the nanoparticle surface. Evaluation of immune responses toward the transgene studied showed a similar titer response as the free protein. However, when the protein was delivered with a cationic lipid as control, titers increased significantly, particularly for the TZ lipid used, which increased titers 1000-fold. These data provide evidence for continued evaluation of TZ lipids as gene delivery vectors and as potential vaccine adjuvants. Finally, in continuing the evaluation of LBNs to improve gene therapy, an LBN based system was evaluated to deplete anti-AAV8 antibodies. As one of the most promising strategies to deliver transgenes since AAV provides an excellent platform that is unfortunately affected by the presence of anti-viral antibodies. This system, using doxorubicin liposomes coated with recombinant VP1 protein bound to DGS-NTA-Ni lipid or DSPE-PEG2000-Maleimide, failed to deplete circulating antibodies to AAV. However, the results of the experiments carried out shed light on how this strategy might be improved upon at a later time. Finally, in an attempt to better understand the immune targets on AAV, the antibody response toward AAV8 was tested in human samples from deidentified blood donors and compared with that of mice and monkeys treated with the virus. Serum from these species was scrutinized for its ability to neutralize the virus in vitro and evaluated using a peptide array for targets against the viral capsid protein VP1. Collectively, the studies presented in the body of this dissertation demonstrate the utility of LBNs in gene delivery, both as vectors and as aids for viral delivery

Fungi in Toe Nails**From the Department of Dermatology, University of Miami School of Medicine, Miami, Florida 33136.

One hundred and eighty three abnormal toe nails were cultured for dermatophytes, molds, and yeasts. Incidence figures for all above mentioned organisms were obtained. In approximately 25% of the abnormal toe nails, dermatophytes were isolated. Trichophyton rubrum and T. mentagrophytes were equally isolated. In approximately 25% of abnormal nails, no organisms were isolated. Non-dermatophytic fungi were commonly associated with abnormal nails (50%). Among these were: Arthroderma quadrificum; Aspergillus sydowii; A. nidulans; Cephalosporium acremonium; Curvularia lunata; Fusarium oxysporum; Hormodendrum cladosporidides; Penicillium citrinum; Scopulariopsis brevicalis.Yeasts were also isolated from 25% of nails (in combination with other organisms). Candida parapsilosis was by far the most commonly yeast isolated. Approximately one half of dermatophytes were recovered by cultural methods when compared to the number of KOH positive nails

Analisis Pengaruh Penyaluran Kredit Usaha Rakyat (KUR) Terhadap Kinerja Usaha Mikro dan Kecil (UMK) Di Kota Payakumbuh

Penelitian ini bertujuan untuk mengetahui dampak penyaluran Kredit Usaha Rakyat (KUR) terhadap modal, tenaga kerja, omzet penjualan, dan keuntungan Usaha Mikro dan Kecil (UMK) di Kota Payakumbuh. Penelitian ini menggunakan data primer yang dilakukan dengan menggunakan metode survey, informasi dikumpulkan dari responden dengan menggunakan kuisioner dan wawancara langsung. Metode yang digunakan dalam penelitian ini adalah metode analisis uji beda rata-rata dengan sampel berpasangan yang digunakan untuk menguji dua data yang berasal dari satu sampel dengan menggunakan uji Wilcoxon Test. Hasil Penelitian ini menunjukan bahwa terdapat perbedaan rata-rata yang signifikan baik pada modal, tenaga kerja, omzet penjualan, dan keuntungan sebelum dan sesudah mendapatkan kredit usaha rakyat (KUR). Program Kredit Usaha Rakyat (KUR) ini hendaknya terus dijalankan dan ditingkatkan, karena penelitian ini membuktikan bahwa program tersebut bisa bermanfaat untuk pengusaha mikro dan kecil di Kota Payakumbuh. Kata kunci : Usaha Mikro dan Kecil, Kredit Usaha Rakyat, Modal, Tenaga Kerja, Omzet Penjualan, dan Keuntungan

High speed drives review: machines, converters and applications

The development of new power electronic device and high performance magnetic materials are the main technological factors that have led both industries and research community to focus their attention on high speed electrical drives. Several papers have already outlined the electrical machine and/or converter topology choice for certain high speed application. This choice depends on the applications under study. This paper aims to identify the most important high speed applications. For each of them, the main design challenges are highlighted and an overview of available market product is presented

Tuberous Sclerosis Complex Activity Is Required to Control Neuronal Stress Responses in an mTOR-Dependent Manner

Tuberous Sclerosis Complex (TSC) is a neurogenetic disorder caused by loss-of-function mutations in either the TSC1 or TSC2 genes and frequently results in prominent CNS manifestations including epilepsy, mental retardation, and autism spectrum disorder. The TSC1/TSC2 protein complex plays a major role in controlling the Ser/Thr kinase mTOR, which is a master regulator of protein synthesis and cell growth. In this study, we show that endoplasmic reticulum (ER) stress regulates TSC1/TSC2 complex to limit mTOR activity. In addition, Tsc2-deficient rat hippocampal neurons and brain lysates from a Tsc1-deficient mouse model both demonstrate elevated ER and oxidative stress. In Tsc2-deficient neurons, the expression of stress markers such as CHOP and HO-1 is increased, and this increase is completely reversed by the mTOR inhibitor rapamycin both in vitro and in vivo. Neurons lacking a functional TSC1/TSC2 complex have increased vulnerability to ER stress-induced cell death via the activation of the mitochondrial death pathway. Importantly, knockdown of CHOP reduces oxidative stress and apoptosis in Tsc2-deficient neurons. These observations indicate that ER stress modulates mTOR activity through the TSC protein complex and that ER stress is elevated in cells lacking this complex. They also suggest that some of the neuronal dysfunction and neurocognitive deficits seen in TSC patients may be due to ER and oxidative stress, and therefore potentially responsive to agents moderating these pathways

Restoring Study 329: efficacy and harms of paroxetine and imipramine in treatment of major depression in adolescence

Objectives: To reanalyse SmithKline Beecham’s Study 329 (published by Keller and colleagues in 2001), the primary objective of which was to compare the efficacy and safety of paroxetine and imipramine with placebo in the treatment of adolescents with unipolar major depression. The reanalysis under the restoring invisible and abandoned trials (RIAT) initiative was done to see whether access to and reanalysis of a full dataset from a randomised controlled trial would have clinically relevant implications for evidence based medicine. Design: Double blind randomised placebo controlled trial. Setting: 12 North American academic psychiatry centres, from 20 April 1994 to 15 February 1998. Participants: 275 adolescents with major depression of at least eight weeks in duration. Exclusion criteria included a range of comorbid psychiatric and medical disorders and suicidality. Interventions: Participants were randomised to eight weeks double blind treatment with paroxetine (20-40 mg), imipramine (200-300 mg), or placebo. Main outcome measures: The prespecified primary efficacy variables were change from baseline to the end of the eight week acute treatment phase in total Hamilton depression scale (HAM-D) score and the proportion of responders (HAM-D score ≤8 or ≥50% reduction in baseline HAM-D) at acute endpoint. Prespecified secondary outcomes were changes from baseline to endpoint in depression items in K-SADS-L, clinical global impression, autonomous functioning checklist, self-perception profile, and sickness impact scale; predictors of response; and number of patients who relapse during the maintenance phase. Adverse experiences were to be compared primarily by using descriptive statistics. No coding dictionary was prespecified. Results: The efficacy of paroxetine and imipramine was not statistically or clinically significantly different from placebo for any prespecified primary or secondary efficacy outcome. HAM-D scores decreased by 10.7 (least squares mean) (95% confidence interval 9.1 to 12.3), 9.0 (7.4 to 10.5), and 9.1 (7.5 to 10.7) points, respectively, for the paroxetine, imipramine and placebo groups (P=0.20). There were clinically significant increases in harms, including suicidal ideation and behaviour and other serious adverse events in the paroxetine group and cardiovascular problems in the imipramine group. Conclusions: Neither paroxetine nor high dose imipramine showed efficacy for major depression in adolescents, and there was an increase in harms with both drugs. Access to primary data from trials has important implications for both clinical practice and research, including that published conclusions about efficacy and safety should not be read as authoritative. The reanalysis of Study 329 illustrates the necessity of making primary trial data and protocols available to increase the rigour of the evidence base.Joanna Le Noury, John M Nardo, David Healy, Jon Jureidini, Melissa Raven, Catalin Tufanaru, Elia Abi-Jaoud

Study 329 continuation phase:Safety and efficacy of paroxetine and imipramine in extended treatment of adolescent major depression

OBJECTIVE: This is an analysis of the unpublished continuation phase of Study 329, the primary objective of which was to compare the efficacy and safety of paroxetine and imipramine with placebo in the treatment of adolescents with unipolar major depression. The objectives of the continuation phase were to assess safety and relapse rates in the longer term. The objective of this publication, under the Restoring Invisible and Abandoned Trials (RIAT) initiative, was to see whether access to and analysis of the previously unpublished dataset from the continuation phase of this randomized controlled trial would have clinically relevant implications for evidence-based medicine. METHODS: The study was an eight-week double-blind randomized placebo-controlled trial with a six month continuation phase. The setting was 12 North American academic psychiatry centres, from 20 April 1994 to 15 February 1998. 275 adolescents with major depression were originally enrolled in Study 329, with 190 completing the eight-week acute phase. Of these, 119 patients (43%) entered the six-month continuation phase (paroxetine n = 49; imipramine n = 39; placebo n = 31), in which participants were continued on their current treatment, blinded. As per the protocol, we have looked at rates of relapse (based on Hamilton Depression Scale scores) across both acute and continuation phases, and generated a safety profile for paroxetine and imipramine compared with placebo for up to six months. ANOVA testing (generalized linear model) using a model including effects of site, treatment and site x treatment interaction was applied. Otherwise we used only descriptive statistics. RESULTS: Of patients entering the continuation phase, 15 of 49 for paroxetine (31%), 12 of 39 for imipramine (31%) and 12 of 31 for placebo (39%) completed as responders. Across the study, 25 patients on paroxetine relapsed (41% of those showing an initial response), 15 on imipramine (26%), and 10 on placebo (21%). In the continuation and taper phases combined there were 211 adverse events in the paroxetine group, 147 on imipramine and 100 on placebo. The taper phase had a higher proportion of severe adverse events per week of exposure than the acute phase, with the continuation phase having the fewest events. CONCLUSIONS: The continuation phase did not offer support for longer-term efficacy of either paroxetine or imipramine. Relapse and adverse events on both active drugs open up the risks of a prescribing cascade. The previously largely unrecognised hazards of the taper phase have implications for prescribing practice and need further exploration

Gender disparities in colloquium speakers at top universities

Colloquium talks at prestigious universities both create and reflect academic researchers' reputations. Gender disparities in colloquium talks can arise through a variety of mechanisms. The current study examines gender differences in colloquium speakers at 50 prestigious US colleges and universities in 2013-2014. Using archival data, we analyzed 3,652 talks in six academic disciplines. Men were more likely than women to be colloquium speakers even after controlling for the gender and rank of the available speakers. Eliminating alternative explanations (e.g., women declining invitations more often than men), our follow-up data revealed that female and male faculty at top universities reported no differences in the extent to which they (i) valued and (ii) turned down speaking engagements. Additional data revealed that the presence of women as colloquium chairs (and potentially on colloquium committees) increased the likelihood of women appearing as colloquium speakers. Our data suggest that those who invite and schedule speakers serve as gender gatekeepers with the power to create or reduce gender differences in academic reputations

Morphological profile of elite brazilian bodybuilders in competitive season

O objetivo deste estudo foi analisar o perfil morfológico de culturistas brasileiros de elite que competiram no XXVII Campeonato Brasileiro de Culturismo-Musculação. Um total de 28 atletas (27,0 ± 7,4 anos; 78,2 ± 8,7kg; 170,1 ± 6,9cm), do sexo masculino, foram voluntariamente estudados. Medidas antropométricas (massa corporal, estatura, espessura de dobras cutâneas, perímetros e diâmetros) foram coletadas de todos os sujeitos no período da manhã, no dia da competição. Os valores estimados da composição corporal dos atletas foram os seguintes: gordura corporal relativa, 5,8 ± 0,5%; massa corporal magra, 73,6 ± 8,2kg; massa muscular, 54,3 ± 6,7kg; área muscular do braço, 108,51 ± 15,19cm². Por outro lado, o somatótipo médio encontrado foi: endomorfia, 1,11 ± 0,29; mesomorfia, 8,26 ± 0,67; ectomorfia, 0,95 ± 0,53 (mesomorfo balanceado). Além de os culturistas apresentarem elevados níveis de desenvolvimento muscular (~70%) e baixos depósitos de gordura corporal, verificou-se uniformidade nos acúmulos de gordura localizada. A predominância do componente mesomorfo confirmou a estrutura muscular acentuadamente desenvolvida encontrada nos atletas investigados. Portanto, os resultados do presente estudo demonstram que atletas de culturismo de elite apresentam no dia da competição grande volume muscular, bem como excelente definição muscular, proporcionalidade e simetria.The aim of this study was to analyze the morphological profile of elite Brazilian bodybuilders who participated in the XXVII Brazilian Championship of Body Building. A total of 28 male athletes (27.0 ± 7.4 years; 78.2 ± 8.7 kg; 170.1 ± 6.9cm), were voluntarily assessed. Anthropometric measures (body mass, stature, skinfolds thickness, perimeters and diameters) were collected from all subjects in the morning shift, at the competition day. The estimated values of the body composition of the athletes were the following: relative body fat, 5.8 ± 0.5%; lean body mass, 73.6 ± 8.2 kg; muscular mass, 54.3 ± 6.7 kg and arm muscular area, 108.51 ± 15.19 cm². Conversely, the mean somatotype found was: endomorphy, 1.11 ± 0.29; mesomorphy, 8.26 ± 0.67; ectomorphy, 0.95 ± 0.53 (balanced mesomorphy). Besides high levels of muscular development (~70%) and low body fat buildup, the bodybuilders also presented uniformity in the accumulation of located fat. Predominance of the mesophormic component confirmed muscular structure remarkably developed found in the investigated athletes. Therefore, the results of the present study demonstrate that elite body building athletes present large amount of muscular volume, as well as excellent muscular definition, proportionality and symmetry at the day of the competition

Challenging SO(10) SUSY GUTs with family symmetries through FCNC processes

We perform a detailed analysis of the SO(10) SUSY GUT model with D3 family symmetry of Dermisek and Raby (DR). The model is specified in terms of 24 parameters and predicts, as a function of them, the whole MSSM set of parameters at low energy scales. Concerning the SM subset of such parameters, the model is able to give a satisfactory description of the quark and lepton masses, of the PMNS matrix and of the CKM matrix. We perform a global fit to the model, including flavour changing neutral current (FCNC) processes Bs --> mu+ mu-, B --> Xs gamma, B --> Xs l+ l- and the B(d,s) - bar B(d,s) mass differences Delta M(d,s) as well as the flavour changing (FC) process B+ --> tau+ nu. These observables provide at present the most sensitive probe of the SUSY mass spectrum and couplings predicted by the model. Our analysis demonstrates that the simultaneous description of the FC observables in question represents a serious challenge for the DR model, unless the masses of the scalars are moved to regions which are problematic from the point of view of naturalness and probably beyond the reach of the LHC. We emphasize that this problem could be a general feature of SUSY GUT models with third generation Yukawa unification and weak-scale minimal flavour violation.Comment: 1 + 37 pages, 5 figures, 11 tables. v3: minor typos fixed. Matches JHEP published versio