Engineering of recombatinant antimalria antibodies for application in paratransgenesis

Malaria, transmitted by Anopheles mosquitoes is responsible for millions of deaths worldwide, especially in the developing countries. The emergence of drug resistant parasites and insecticide resistant vectors has stimulated efforts to develop novel genetic strategies to modify the insect vector and reduce its competence to transmit the parasite. One proposed approach is the genetic manipulation of insect’s midgut symbionts to express anti-parasite molecules. Recombinant antibodies that target specific antigens expressed on the parasites’ surface could be used as anti-parasite molecules, especially if they could not only bind but agglutinate the target. The murine antibody 4B7 binds to Pfs25 epitope expressed on the zygote and ookinette stages of the parasite. Pf-NPNA- 1 is a human antibody that specifically binds to the NPNA (Asn-Pro-Asn-Ala) repeats of the circumsporozoite protein expressed on the sporozoite stage of the malaria parasite. This study aimed to characterise these antibodies for their application in symbiont control. For this purpose, the antibodies have been codon optimised for bacterial expression and formatted as single chain variable fragments (scFv). Synthetic genes encoding the scFv 4B7 and Pf-NPNA-1 were constructed, with varying linker length, in the VH-VL and VL-VH orientation. The scFvs were cloned into different expression plasmids to evaluate a suitable expression system. The orientation of the variable domains on secretion of the scFv 4B7 was investigated. No secretion was observed for the scFv 4B7 in the VH-VL orientation. For the reverse orientation, scFv 4B7 (VL-VH) was poorly secreted with no antigen binding. Secretion was observed for a variant of scFv 4B7 but this did not show significant antigen binding. Pf-NPNA-1 scFv constructs, in the VH-VL orientation, were efficiently secreted and showed detectable binding to antigen. Multimeric assembly of the scFv constructs was evaluated by varying the linker length. 4B7 and Pf-NPNA-1 scFv constructs exhibited monomeric, dimeric and multimeric assembly. Fusion of the human kappa constant domain to the scFvs resulted in formation of monomeric and higher ordered forms. Transfer of the scFv gene fragments into a broad-host vector facilitated evalution of recombinant antibody expression in the acetic acid bacterium, Asaia SF2.1. In summary, the results from this study demonstrate the potential utility of the antibodies, 4B7 and Pf-NPNA-1, as anti-parasite molecules for blockade of malaria transmission via mosquito midgut symbionts

Network estimation in State Space Model with L1-regularization constraint

Biological networks have arisen as an attractive paradigm of genomic science ever since the introduction of large scale genomic technologies which carried the promise of elucidating the relationship in functional genomics. Microarray technologies coupled with appropriate mathematical or statistical models have made it possible to identify dynamic regulatory networks or to measure time course of the expression level of many genes simultaneously. However one of the few limitations fall on the high-dimensional nature of such data coupled with the fact that these gene expression data are known to include some hidden process. In that regards, we are concerned with deriving a method for inferring a sparse dynamic network in a high dimensional data setting. We assume that the observations are noisy measurements of gene expression in the form of mRNAs, whose dynamics can be described by some unknown or hidden process. We build an input-dependent linear state space model from these hidden states and demonstrate how an incorporated $L_{1}$ regularization constraint in an Expectation-Maximization (EM) algorithm can be used to reverse engineer transcriptional networks from gene expression profiling data. This corresponds to estimating the model interaction parameters. The proposed method is illustrated on time-course microarray data obtained from a well established T-cell data. At the optimum tuning parameters we found genes TRAF5, JUND, CDK4, CASP4, CD69, and C3X1 to have higher number of inwards directed connections and FYB, CCNA2, AKT1 and CASP8 to be genes with higher number of outwards directed connections. We recommend these genes to be object for further investigation. Caspase 4 is also found to activate the expression of JunD which in turn represses the cell cycle regulator CDC2.Comment: arXiv admin note: substantial text overlap with arXiv:1308.359

High dimensional Sparse Gaussian Graphical Mixture Model

This paper considers the problem of networks reconstruction from heterogeneous data using a Gaussian Graphical Mixture Model (GGMM). It is well known that parameter estimation in this context is challenging due to large numbers of variables coupled with the degeneracy of the likelihood. We propose as a solution a penalized maximum likelihood technique by imposing an $l_{1}$ penalty on the precision matrix. Our approach shrinks the parameters thereby resulting in better identifiability and variable selection. We use the Expectation Maximization (EM) algorithm which involves the graphical LASSO to estimate the mixing coefficients and the precision matrices. We show that under certain regularity conditions the Penalized Maximum Likelihood (PML) estimates are consistent. We demonstrate the performance of the PML estimator through simulations and we show the utility of our method for high dimensional data analysis in a genomic application

PENGARUH TERAPI MUSIK JAWA TERHADAP INSOMNIA PADA LANSIA DI RW 01 DESA PAGERTANJUNG PLOSO JOMBANG

Sebagian besar lansia mengaku kesulitan untuk memulai tidur, sering terbangun tengah malam, dan sulit untuk kembali tidur, tidur tidak tenang dan terbangun lebih awal. Terapi musik Jawa merupakan alternatif untuk mengatasi masalah tersebut. Tujuan penelitian ini adalah untuk mengetahui pengaruh terapi musik Jawa terhadap insomnia pada lansia di RW 01 Desa Pagertanjung Ploso Jombang. Jenis penelitian menggunakan Pre-Experimental yaitu One group pra-post test design. Populasinya seluruh lansia berusia 60 tahun ke atas yang mengalami insomnia, tidak mengalami gangguan fisik serta menyukai musik Jawa sebesar 34 lansia. Besar sampel 31 lansia dengan teknik simple random sampling. Variabel independen adalah terapi musik Jawa dengan MP3 dan dependen adalah insomnia menggunakan instrumen SOP dan kuesioner. Uji statistik menggunakan uji McNemar Test dengan α = 0,005. Hasil penelitian menunjukkan sebelum diberi terapi musik Jawa seluruhnya (100%) mengalami insomnia, setelah diberi terapi musik Jawa sebagian besar (64,5%) tidak mengalami insomnia. Hasil uji statistik didapatkan nilai p (0,000) berarti α (0,005) > p, dengan demikian H0 ditolak yang artinya ada pengaruh terapi musik Jawa terhadap insomnia pada lansia di RW 01 Desa Pagertanjung Ploso Jombang. Pemberian terapi musik Jawa berpengaruh terhadap insomnia sehingga dapat mencegah ataupun menghindari insomnia semakin parah. Maka, lansia diharapkan secara rutin mendengarkan musik Jawa sebelum tidur

Fault identification-based voltage sag state estimation using artificial neural network

This paper presents an artificial neural network (ANN) based approach to identify faults for voltage sag state estimation. Usually ANN cannot be used to abstract relationship between monitored data and arbitrarily named fault indices which are not related at all logically in numerical level. This paper presents a novel approach to overcome this problem. In this approach, not only the networks are trained to adapt to the given training data, the training data (the expected outputs of fault indices) is also updated to adapt to the neural network. During the training procedure, both the neural networks and training data are updated interactively. With the proposed approach, various faults can be accurately identified using limited monitored data. The approach is robust to measurement uncertainty which usually exists in practical monitoring systems. Furthermore, the updated fault indices are able to suggest the difference of the impact of various faults on bus voltages

Integrated care programmes for adults with chronic conditions: a meta-review

Objective To review systematic reviews and meta-analyses of integrated care programmes in chronically ill patients, with a focus on methodological quality, elements of integration assessed and effects reported. Design Meta-review of systematic reviews and meta-analyses identified in Medline (1946-March 2012), Embase (1980-March 2012), CINHAL (1981-March 2012) and the Cochrane Library of Systematic Reviews (issue 1, 2012). Main Outcome Measures Methodological quality assessed by the 11-item Assessment of Multiple Systematic Reviews (AMSTAR) checklist; elements of integration assessed using a published list of 10 key principles of integration; effects on patient-centred outcomes, process quality, use of healthcare and costs. Results Twenty-seven systematic reviews were identified; conditions included chronic heart failure (CHF; 12 reviews), diabetes mellitus (DM; seven reviews), chronic obstructive pulmonary disease (COPD; seven reviews) and asthma (five reviews). The median number of AMSTAR checklist items met was five: few reviewers searched for unpublished literature or described the primary studies and interventions in detail. Most reviews covered comprehensive services across the care continuum or standardization of care through inter-professional teams, but organizational culture, governance structure or financial management were rarely assessed. A majority of reviews found beneficial effects of integration, including reduced hospital admissions and re-admissions (in CHF and DM), improved adherence to treatment guidelines (DM, COPD and asthma) or quality of life (DM). Few reviews showed reductions in costs. Conclusions Systematic reviews of integrated care programmes were of mixed quality, assessed only some components of integration of care, and showed consistent benefits for some outcomes but not other

Retrospective checking of compliance with practice guidelines for acute stroke care: a novel experiment using openEHR’s Guideline Definition Language

BACKGROUND: Providing scalable clinical decision support (CDS) across institutions that use different electronic health record (EHR) systems has been a challenge for medical informatics researchers. The lack of commonly shared EHR models and terminology bindings has been recognised as a major barrier to sharing CDS content among different organisations. The openEHR Guideline Definition Language (GDL) expresses CDS content based on openEHR archetypes and can support any clinical terminologies or natural languages. Our aim was to explore in an experimental setting the practicability of GDL and its underlying archetype formalism. A further aim was to report on the artefacts produced by this new technological approach in this particular experiment. We modelled and automatically executed compliance checking rules from clinical practice guidelines for acute stroke care. METHODS: We extracted rules from the European clinical practice guidelines as well as from treatment contraindications for acute stroke care and represented them using GDL. Then we executed the rules retrospectively on 49 mock patient cases to check the cases’ compliance with the guidelines, and manually validated the execution results. We used openEHR archetypes, GDL rules, the openEHR reference information model, reference terminologies and the Data Archetype Definition Language. We utilised the open-sourced GDL Editor for authoring GDL rules, the international archetype repository for reusing archetypes, the open-sourced Ocean Archetype Editor for authoring or modifying archetypes and the CDS Workbench for executing GDL rules on patient data. RESULTS: We successfully represented clinical rules about 14 out of 19 contraindications for thrombolysis and other aspects of acute stroke care with 80 GDL rules. These rules are based on 14 reused international archetypes (one of which was modified), 2 newly created archetypes and 51 terminology bindings (to three terminologies). Our manual compliance checks for 49 mock patients were a complete match versus the automated compliance results. CONCLUSIONS: Shareable guideline knowledge for use in automated retrospective checking of guideline compliance may be achievable using GDL. Whether the same GDL rules can be used for at-the-point-of-care CDS remains unknown

Digital tanlock loop architecture with no delay

This article proposes a new architecture for a digital tanlock loop which eliminates the time-delay block. The �=2 (rad) phase shift relationship between the two channels, which is generated by the delay block in the conventional timedelay digital tanlock loop (TDTL), is preserved using two quadrature sampling signals for the loop channels. The proposed system outperformed the original TDTL architecture, when both systems were tested with frequency shift keying input signal. The new system demonstrated better linearity and acquisition speed as well as improved noise performance compared with the original TDTL architecture. Furthermore, the removal of the time-delay block enables all processing to be digitally performed, which reduces the implementation complexity. Both the original TDTL and the new architecture without the delay block were modelled and simulated using ATLAB/Simulink. Implementation issues, including complexity and relation to simulation of both architectures, are also addressed

Performance evaluation of the time delay digital tanlock loop architectures

This article presents the architectures, theoretical analyses and testing results of modified time delay digital tanlock loop (TDTLs) system. The modifications to the original TDTL architecture were introduced to overcome some of the limitations of the original TDTL and to enhance the overall performance of the particular systems. The limitations addressed in this article include the non-linearity of the phase detector, the restricted width of the locking range and the overall system acquisition speed. Each of the modified architectures was tested by subjecting the system to sudden positive and negative frequency steps and comparing its response with that of the original TDTL. In addition, the performance of all the architectures was evaluated under noise-free as well as noisy environments. The extensive simulation results using MATLAB/SIMULINK demonstrate that the new architectures overcome the limitations they addressed and the overall results confirmed significant improvements in performance compared to the conventional TDTL system