Double Fusion Research Articles

Intramedullary nail versus external fixator for ankle arthrodesis in Charcot neuroarthropathy: A meta-analysis of comparative studies.

Ankle Charcot neuroarthropathy presents a great challenge for treatment. While conservative treatments yield poor results, arthrodesis is considered the treatment of choice for unstable Charcot ankles. There are two methods used for ankle fusion: retrograde intramedullary nail (IMN) and external fixator (EF). Literature reports inconclusive results on which method is better for arthrodesis. The aim of this meta-analysis is to look for significant difference between the two techniques in terms of fusion rate and complications. Four comparative studies including 117 patients (117 interventions) were located and analyzed. IMN was found to yield double fusion rate than EF ( p = 0.2) with 5 weeks less time to heal ( p = 0.4). While no significance was noted for revision surgery and amputation rates, EF yielded higher rates of hardware infection ( p = 0.01) and wound infection ( p = 0.01). The IMN method seems to offer quicker and better fusion rate when compared to EF with significantly lower postoperative infection rates. In the clinical context of high-risk patients such as the diabetic population, IMN technique could be a better option in the case of ulcer-free ankles/feet or in the presence of medial/lateral ulcers. In case of presence of plantar ulcers, EF might be a better choice to avoid the plantar approach for nail insertion. Future research with bigger sample sized and randomized studies are warranted for validation.

Automatic detection of passable roads after floods in remote sensed and social media data

This paper addresses the problem of floods classification and floods aftermath detection based on both social media and satellite imagery. Automatic detection of disasters such as floods is still a very challenging task. The focus lies on identifying passable routes or roads during floods. Two novel solutions are presented, which were developed for two corresponding tasks at the MediaEval 2018 benchmarking challenge. The tasks are (i) identification of images providing evidence for road passability and (ii) differentiation and detection of passable and non-passable roads in images from two complementary sources of information. For the first challenge, we mainly rely on object and scene-level features extracted through multiple deep models pre-trained on the ImageNet and Places datasets. The object and scene-level features are then combined using early, late and double fusion techniques. To identify whether or not it is possible for a vehicle to pass a road in satellite images, we rely on Convolutional Neural Networks and a transfer learning-based classification approach. The evaluation of the proposed methods is carried out on the large-scale datasets provided for the benchmark competition. The results demonstrate significant improvement in the performance over the recent state-of-art approaches.

Bax to the future – A novel, high-yielding approach for purification and expression of full-length Bax protein for structural studies

Mitochondria-mediated apoptosis (programmed cell death) involves a sophisticated signaling and regulatory network that is regulated by the Bcl-2 protein family. Members of this family have either pro- or anti-apoptotic functions. An important pro-apoptotic member of this family is the cytosolic Bax. This protein is crucial for the onset of apoptosis by perforating the mitochondrial outer membrane (MOM). This process can be seen as point of no return, since disintegration of the MOM leads to the release of apotogenic factors such as cytochrome c into the cytosol triggering the activation of caspases and subsequent apoptotic steps. Bax is able to interact with the MOM with both its termini, making it inherently difficult to express in E. coli. In this study, we present a novel approach to express and purify full-length Bax with significantly increased yields, when compared to the commonly applied strategy. Using a double fusion approach with an N-terminal GST-tag and a C-terminal Intein-CBD-tag, we were able to render both Bax termini inactive and prevent disruptive interactions from occurring during gene expression. By deploying an Intein-CBD-tag at the C-terminus we were further able to avoid the introduction of any artificial residues, hence ensuring the native like activity of the membrane-penetrating C-terminus of Bax. Further, by engineering a His6-tag to the C-terminus of the CBD-tag we greatly improved the robustness of the purification procedure. We report yields for pure, full-length Bax protein that are increased by an order of magnitude, when compared to commonly used Bax expression protocols.

An improved energy efficient cluster head selection protocol using the double cluster heads and data fusion methods for IoT applications

The more energy consumption is the major issue for wireless sensor network. In wireless sensor network (WSN), sensor nodes are fixed in various routing algorithms. In the same networks, mobile sensor and fixed sensor nodes are combine and its used in few applications. The function is corruption while mobility was achieved, since these nodes have minimum battery power, lower range of communication and a lessor amount of memory. To overcome this issue, Improved Energy Efficient Cluster Head Selection protocol (IEECHS-WSN) is proposed, in these technique is used to transfer the received information by using energy efficient routing protocol. In the CH election method, two cluster heads are selected in a separated cluster and its work in various functions, this can be prolong the network lifetime and decrease the energy consumption of IoT applications. Proposed technique is described on clustering of dual CHs in the method of data fusion for data entropy. This information entropy is used for fusion and classification, the result of fusion and classification are accurate and efficient for data transmission. Our proposed IEECHS protocol has better throughput, lifetime of network and energy consumption compared than the existing technique.

A prospective randomized study to analyze the efficacy of balanced pre-emptive analgesia in spine surgery

Background ContextSurgical procedures involving the spine are known to cause moderate to severe postoperative pain. Inadequate management of acute pain in the postoperative period results in higher morbidity, and consequently may lead to chronic pain caused by central sensitization. The role of pre-emptive analgesia (PA) and intraoperative analgesia in management of postoperative pain has gained precedence over recent years. Pathophysiology of postoperative pain in spine surgery is unique, as it is a combination of nociceptive, inflammatory, and neuronal stimuli. Blockage of all three stimuli in the perioperative period by pre-emptively administrating a combination of paracetamol (P), ketorolac (K), and pregabalin (PR) might help in adequate management and alleviation of acute postoperative pain. PurposeTo evaluate the analgesic effect of a combination of P, K, and PR as pre-emptive multimodal analgesia, aimed to block or reduce acute postoperative pain after spine surgery. Study DesignA prospective, randomized, controlled, and double-blinded clinical trial. Patient SampleAfter Institutional Review Board approval, 100 consecutive patients requiring single- or double level spinal fusion procedures were randomized into two groups—PA and control (C). MethodsThe PA group received P, K, and PR 4 hours before surgery, as PA. Both groups underwent identical anesthetic and postoperative pain management protocol. Outcome MeasuresDemographic and surgical data, 4 hourly postoperative pain levels—Numeric Pain Rating scale (NRS), Ambulatory NRS scores; level of consciousness—Ramsay sedation scale, total amount of opioids consumed (TOC) through patient-controlled analgesia; functional levels—Oswestry Disability Index (ODI), surgical satisfaction index-North American Spine Society (NASS) satisfaction scale, duration of hospital stay, and all complications were recorded and analyzed. A research grant of 6,032 USD was obtained from AO Spine toward this work. There is no conflict to disclose. ResultsBoth the groups had identical demographic backgrounds and surgical profiles. The average NRS score within the first 48-hour period in the PA group (2.7±0.79) was significantly less than the C group (3.4±0.98) and the differences were more in the first 12 hours following surgery. Similarly, Ambulatory NRS scores were significantly low in the PA group during the first and second postoperative days. The PA group individuals were found to be more physically motivated, as 95.7% were able to ambulate 50 m on the first postoperative day compared with 30% in the C group. The PA group had significantly low TOC (3.02±2.29 mg) in comparison to the C group (4.94±3.08 mg). The duration of hospital stay was 4.17±1.02 and 4.84±1.62 days in the PA and C groups (p=.017), respectively. No major complications occurred in either groups and were found to be similar in percentage between both the groups, except for nausea and vomiting which were more in C group. Dry mouth was the most common side effect noted irrespective of the groups. All patients had significant improvement in ODI with better results in PA group at first month follow up. The PA group (97.90%) was extremely satisfied compared with C group (72%, p=.002) according to NASS scale. ConclusionsPostoperative pain management in spine surgery is maximized if perioperative painful stimuli can be inhibited, which requires adequate blood levels of analgesic, anti-inflammatory, and neuropathic drugs intraoperatively. The employed strategy of preoperative administration of balanced analgesia with a combination of P, K, and PR, each having different mechanisms of action, resulted in lesser pain intensity, allowed better ambulation tolerance, improved functional outcomes and has also reduced the requirement of opioids and duration of hospital stay with no additional complications. Thus, this balanced analgesia administered preoperatively would address the complicated postsurgical pain.

Military Object Real-Time Detection Technology Combined with Visual Salience and Psychology

This paper presents a method of military object detection through the combination of human visual salience and visual psychology, so as to achieve rapid and accurate detection of military objects on the vast and complex battlefield. Inspired by the process of human visual information processing, this paper establishes a salient region detection model based on double channel and feature fusion. In this model the pre-attention channel is to process information on the position and contrast of images, and the sub-attention channel is to integrate information on primary visual features first and then merges results of the two channels to determine the salient region. The main theory of Gestalt visual psychology is then used as the constraint condition to integrate the candidate salient regions and to obtain the object figure with overall perception. After that, the efficient sub-window search method is used to detect and filter the object in order to determine the location and range of objects. The experimental results show that, when compared with the existing algorithms, the algorithm proposed in this paper has prominent advantages in precision, effectiveness, and simplicity, which not only significantly reduces the effectiveness of battlefield camouflage and deception but also achieves the rapid and accurate detection of military objects, thus promoting its application prospect.

Friday, September 28, 2018 1:00 PM–2:30 PM abstracts: achieving lumbar interbody fusion: 192. Clinical evaluation of a new allograft cage designed for TLIF: a minimal 1-year follow-up

BACKGROUND CONTEXT Currently most cages are made of metal, carbon fiber, polyether ether ketone (PEEK), and bio-absorbable materials. But there are some drawbacks of these materials in terms of biology, biomechanics, etc., mainly in terms of the elastic modulus is too large, poor biocompatibility, and unable to absorb. Allogeneic bone is recognized bone graft material of great osteogenic effect. It has the closest physicochemical properties, structure and activity of protein-based composition of autologous bone. It in rich in a variety of cytokines required for osteogenesis: bone morphogenetic protein-2, vascular endothelial growth factor, core binding factor α1, et. PURPOSE The purpose of this study was to observe the preliminary clinical results of the new allograft cage. STUDY DESIGN/SETTING A retrospective research. PATIENT SAMPLE A total of 152 patients. OUTCOME MEASURES Visual Analogue Scale (VAS) and The Oswestry Disability Index (ODI);Relative intervertebral disc height (RIDH); fusion rate. METHODS From 2012 to 2015, a retrospective review of the first 152 consecutive cases using the new allograft cage in our department was performed. The follow-up ranged from 12 to 36 months (average 18 months). Intraoperative index, clinical outcome and complications were recorded. Radiographs evaluatedrelative intervertebral disc height (RIDH) and fusion rate. RESULTS Between September 2012 and November 2014, total 152 patients surgery using the new allograft cage in our department. A total of 133 c (87.6%) met the inclusion criteria and exclusion criteria. There are 78 males and 55 females. The average follow-up time is 15.2±5.3 months. The main diagnosis are lumbar disc herniation (68/133), Lumbar spinal stenosis (44/133), SpondyloliSthesis (21/133). There are 99 one segments fusion and 33 two segments fusion. Respectively, the average blood loss and operation time is 335.7±214.2ml and 175.2±40.9 minutes in single segments fusion, and 488.3±173.2ml, 335.7±214.2 minutes in double segments fusion. The VAS score decreased significantly from 6.89 ±2.51 preoperatively to 2.72±0.97 in 3 months postoperation. At final follow up, the VAS score is 2.96 ±1.02. There was also a significant improvement of the ODI from 38.3±15.9preoperatively to 14.6±6.3 in 3 months postoperation (P CONCLUSIONS It has the potential ability in promoting fusion rate in relative short time (approximately 6 months) postoperatively. We consider it have good clinical outcomes. However, it is just the preliminary data with a short-term follow up. Long-term follow-up is still needed proved.

Does Cervical Plate Add Value in Single or Double Level Anterior Cervical Discectomy and Fusion?

Background Data: Optimal surgical approach for anterior cervical discectomy and fusion (ACDF) is still controversial. Some authors believe that addition of locking plate (ACDFP) has higher costs and longer operative time. Other surgeons believe that ACDFP have better clinical outcomes and more sound fusion. The debate is higher in single and doublelevel ACDF.Purpose: To compare the clinical and radiological outcomes in patients with single or double level cervical disc herniation operated by ACDF versus those operated by ACDFP. Study Design: A retrospective, descriptive controlled, non-randomized, clinical case study. Patients and Methods: Forty patients underwent ACDF. They were divided into twogroups; ACDF group and ACDFP fixation group. Each group included 20 patients. They were operated between 2010 and 2017. Participants were evaluated pre-operatively and post-operatively 1, 3, 6, 9 and 12 months after surgery. Operative time, hospital stay and complications were assessed. Pain was scored by a VAS for both upper arm and cervicalpain. The clinical outcomes were compared using Odom’s criteria. Cervical fusion was assessed radiologically. Results: The demographic data of the two groups of patients were fairly homogeneous and comparable. ACDFP group showed slightly better clinical outcome parameters in comparison to ACDF group including VAS for cervical pain (9±5 versus 28±11) and Odom’s criteria scores (15 excellent outcome versus 10). Also ACDFP group showed slightly better radiological fusion rate (100%) in comparison to (85%) in ACDF group. Reported nonunion was higher in ACDF group (15%) in comparison to ACDFP group (0%). Conclusion: Our data suggest that the addition of cervical plate fixation to ACDF mightleads to better clinical outcome and radiological fusion in single and double cervical disc disease. (2018ESJ155)

Stable Core-Modified Doubly N-Fused Expanded Dibenziporphyrinoids.

We describe one-pot synthesis of stable doubly N-fused expanded dibenziporphyrinoids using readily available precursors under acid-catalyzed conditions. The doubly N-fused expanded dibenziporphyrinoids have been synthesized by adopting an inversion followed by fusion strategy. The studies showed that the dibenziporphyrinoids undergo mono fusion initially, but due to the high stability of doubly fused dibenziporphyrinoids, the monofused macrocycles undergo further fusion to form doubly fused dibenziporphyrinoids. The mono fusion and double fusion in these dibenziporphyrinoids were established by X-ray crystallography.

Dried Jujube Classification Based on a Double Branch Deep Fusion Convolution Neural Network

Dried Jujube Classification Based on a Double Branch Deep Fusion Convolution Neural Network

Evaluation of Different Modalities of Anterior Cervical Discectomy for Treatment of Single and Double Level Cervical Disc Herniation

Introduction: Cervical disc herniation and degenerative disease of the cervical spine are the most common causes of cervical cord and nerve root dysfunction. The surgical treatment of cervical radiculopathy is still controversial. Instead of two possibilities, nowadays three possible treatments concur with each other: anterior cervical discectomy without implantation of any structure, anterior cervical discectomy with fusion, and finally, cervical discectomy with implantation of disc prosthesis. Patients and methods: Twenty patients with cervical disc herniation with radiculopathy, which had not responded to conservative treatment were treated by anterior decompression and cervical disc replacement. All these patients were evaluated preoperatively clinically and radiologically (plain X-rays of cervical spine; A-P, Lateral and dynamic films: flexion, extension and oblique and MRI cervical spine). Nerve conduction study was done to exclude peripheral compression neuropathy and to confirm radiculopathy in selected cases where double entrapment phenomena suspected and followed up for a period of 1 month to 12 months. The technique followed anterior decompression differed according to the way of reconstruction at each level and the patients were accordingly classified into three groups: group A (1-level fusion); where the anterior decompression was accomplished by single level cervical discectomy, and then insertion of cervical cage at this leve, group B (2-level fusion); where the anterior decompression was accomplished by double level cervical discectomy, and then insertion of cervical cage at both levels, group C (hybrid construct); where the anterior decompression was in the form of one level cervical discectomy, followed by cage implantation at this level and another level cervical discectomy followed by insertion of cervical disc prosthesis at the same time. Functional outcome was assessed according to Odom’s criteria. Postoperative plain X-rays of cervical spine (A-P and Lateral) were done at follow-up visits (immediate postoperative, 3 months, 6 months 12 month postoperatively. MRI or CT of the cervical spine is done for patients routinely and for patients not improving or with persistent preoperative complaint or any new neurologic deficit. Results and Discussion: The ages in our patient population ranged from 30 to 50 years, with a mean of 40 ± 5.9 years (mean ± standard deviation). 11/20 patients (55%) were males, and 9/20 (45%) were females. The commonest level affected is C5-C6 level. The most common complaint of patients is neck pain and radiculopathy. Anterior cervical discectomy followed by single level cervical fusion was done on 13 patients, while 5 patients were subjected to anterior cervical discectomy followed by double level cervical fusion and another 2 patients had anterior cervical discectomy followed by cervical artificial disc replacement at one level and zero profile implant insertion and fusion at another level. Regarding the mean duration of hospital stay it was 2.4 days in the single level group while it was 3.8 days in the double level group and 4 days in the hybrid group. Regarding the functional outcome 9 patients (69.2%) had excellent outcome in the single level group versus 3 patients (60%) In the double level group and 2 patients (100%) in the hybrid group, while there were 3 patients (23.1%) who had good outcome in the single level group and 1 patient (20%) in the double level group, finally only one patient in the single level group and 1 patient in the double level group who had satisfactory outcome. There were 2 complications in this study, one (7.7%) in the single level group and one (20%) in the double level group. In the single level group 1 patient had removal of prosthesis due to device failure and hypermobility. In the double level group, one patient had temporary dysphagia and dysphonia. Conclusion: Ideal treatment for cervical degenerative disc disease must deal with and improve its three components (axial neck pain, radiculopathy, and myelopathy), normalize cervical spine biomechanics so not to act as a nidus accelerating the degenerative process, and improves the functional outcome of the patient without serious complications. CDR and ACDF are both effective treatment strategies for managing degenerative conditions of the cervical spine. There is insufficient evidence to show which technique is the most effective and provides the longest-lasting symptom relief.

Porphyrin Arch-Tapes: Synthesis, Contorted Structures, and Full Conjugation.

Porphyrin tapes possessing meso-meso β-β β-β triple direct linkages have been targets of extensive studies because of their fully conjugated characteristic π-electronic networks. In this paper, we report porphyrin arch-tapes that bear additional carbonyl group(s) or methylene group(s) inserted between one of the β-β linkage(s) of the porphyrin tapes. The carbonyl-inserted porphyrin arch-tapes were efficiently synthesized by double fusion reactions of β-to-β carbonyl-bridged porphyrin oligomers with DDQ and Sc(OTf)3, and were converted to the methylene-bridged porphyrin arch-tapes via Luche reduction with NaBH4 and CeCl3 followed by ionic hydrogenation with HBF4·OEt2 and BH3·NEt3. While the conventional porphyrin tapes display rigid and planar structures and low solubilities, these porphyrin arch-tapes show remarkably contorted structures, flexible conformations, and improved solubilities because of the presence of the incorporated seven-membered ring(s). Interestingly, the methylene-inserted arch-tapes exhibited conjugative electronic interactions that were comparable to those of porphyrin tapes probably owing to through-space interaction in the contorted conformations. The carbonyl-inserted arch-tapes displayed distinctly larger conjugative interactions owing to an active involvement of the carbonyl group(s) in the electronic conjugation. A similar trend was observed in the nonlinear optical properties, as evidenced by their two-photon absorption cross sections. Furthermore, as a benefit of the contorted structures, these porphyrin arch-tapes can catch C60 fullerene effectively. Naturally, the electron-rich methylene-bridged arch-tapes exhibited larger association constants than the electron-deficient carbonyl-bridged arch-tapes. Among these arch-tapes, a methylene-bridged syn-Ni(II) porphyrin trimer recorded the largest association constant of (1.5 ± 0.4) × 107 M-1 in toluene at 25 °C.

Sequence-independent cloning methods for long DNA fragments applied to synthetic biology

Simplified methods to assemble DNA fragments by independent cloning sequence have helped in the progress of synthetic biology, allowing some biotechnological processes to become economically viable by genetic improvement of microorganisms. We compared three methods of assembling six DNA fragments: PCR fusion-based, isothermal NEBuilder and circular polymerase extension cloning (CPEC). Double and triple fusion occurs directly with the PCR products using PCR fusion-based and NEBuilder methods. For multiple fragments the results showed higher efficiency by the CPEC method which allowed assembly of six fragments previously purified by agarose gel extraction, after a sequence of 20 annealing/extension cycles without any primer.

A Bayesian Double Fusion Model for Resting-State Brain Connectivity Using Joint Functional and Structural Data.

Current approaches separately analyze concurrently acquired diffusion tensor imaging (DTI) and functional magnetic resonance imaging (fMRI) data. The primary limitation of these approaches is that they do not take advantage of the information from DTI that could potentially enhance estimation of resting-state functional connectivity (FC) between brain regions. To overcome this limitation, we develop a Bayesian hierarchical spatiotemporal model that incorporates structural connectivity (SC) into estimating FC. In our proposed approach, SC based on DTI data is used to construct an informative prior for FC based on resting-state fMRI data through the Cholesky decomposition. Simulation studies showed that incorporating the two data produced significantly reduced mean squared errors compared to the standard approach of separately analyzing the two data from different modalities. We applied our model to analyze the resting state DTI and fMRI data collected to estimate FC between the brain regions that were hypothetically important in the origination and spread of temporal lobe epilepsy seizures. Our analysis concludes that the proposed model achieves smaller false positive rates and is much robust to data decimation compared to the conventional approach.

Clean & Sustainable process for Zeolite Synthesis: Effect of Acidic Pretreatment and Double Fusion on the Characteristics

The application of Coal fly ash (CFA) to synthesize a catalyst finds a noble way to utilize this abundant waste material as their management has been a concern with various approaches for its beneficial uses. Various research and development activities were being conducted to identify and develop the synthesis of catalyst (Zeolite) from coal fly ash (CFA). In this research, CFA was used to synthesize CFA a type of Zeolite catalyst being commercially used, with the help of acid pre-treatment and base post treatment which is subsequently followed by hydrothermal treatment. The use of mixing CFA with acids and bases advances the physicochemical properties of the catalysts such as pore size, pore volume, stability, crystal structure etc. and hence the activity of the Zeolite. The physicochemical properties of Zeolite were evaluated by XRD, XRF and SEM analysis. XRD was used to determine the crystal structure of the Zeolite crystals, SEM was used to determine the morphological structure and texture properties and XRF was used for routine, relatively non-destructive chemical analysis of the Zeolite. The fusion temperature and drying temperature had significant effect in this process of Zeolite preparation. This work also suggests that the performance of the Zeolite obtained from the optimal conditions is more efficient than that of the commercial catalysts. Thus, the aim of this work is to valorize the CFA waste for the preparation of the Zeolite.

Detecting anomalous events in videos by learning deep representations of appearance and motion

Anomalous event detection is of utmost importance in intelligent video surveillance. Currently, most approaches for the automatic analysis of complex video scenes typically rely on hand-crafted appearance and motion features. However, adopting user defined representations is clearly suboptimal, as it is desirable to learn descriptors specific to the scene of interest. To cope with this need, in this paper we propose Appearance and Motion DeepNet (AMDN), a novel approach based on deep neural networks to automatically learn feature representations. To exploit the complementary information of both appearance and motion patterns, we introduce a novel double fusion framework, combining the benefits of traditional early fusion and late fusion strategies. Specifically, stacked denoising autoencoders are proposed to separately learn both appearance and motion features as well as a joint representation (early fusion). Then, based on the learned features, multiple one-class SVM models are used to predict the anomaly scores of each input. Finally, a novel late fusion strategy is proposed to combine the computed scores and detect abnormal events. The proposed ADMN is extensively evaluated on publicly available video surveillance datasets including UCSD pedestian, Subway, and Train, showing competitive performance with respect to state of the art approaches.

Electron Delocalization in Perylene Diimide Helicenes

AbstractWe report two new helicenes derived from the double fusion of an acene with two perylene diimide (PDI) subunits. These PDI‐helicene homologs exhibit very different structural and electronic properties, despite differing by only a single ring in the link between the PDI units. The shorter inter‐PDI link brings the two PDI subunits closer together, and this results in the collision of their respective π‐electron clouds. This collision facilitates intramolecular through‐space electronic delocalization when the PDI‐helicene is reduced.

Electron Delocalization in Perylene Diimide Helicenes.

We report two new helicenes derived from the double fusion of an acene with two perylene diimide (PDI) subunits. These PDI-helicene homologs exhibit very different structural and electronic properties, despite differing by only a single ring in the link between the PDI units. The shorter inter-PDI link brings the two PDI subunits closer together, and this results in the collision of their respective π-electron clouds. This collision facilitates intramolecular through-space electronic delocalization when the PDI-helicene is reduced.

Energy transfer between fusion biliproteins co-expressed with phycobiliprotein in Escherichia coli

In cyanobacteria, phycobiliproteins (PBS) show excellent energy transfer among the chromophores absorbing over most of the visible. The energy transfers are used to study phycobilisome assembly and bioimaging. Using All4261GAF2(C81L) as energy donor, ApcE(1-240/Δ87-130) as energy acceptor, we co-expressed fusion protein ApcE(1-240/Δ87-130)::All4261GAF2(C81L) with phycobiliprotein in Escherichia Coli and studied the energy transfer between two protein domains. With N-terminal His6 tag, ApcE(1-240/Δ87-130)::All4261GAF2(C81L) cannot be purified by nickel-affinity column. We added six histidines in the C-terminal of ApcE(1-240/Δ87-130)::All4261GAF2(C81L) and co-expressed it with phycobiliprotein. ApcE(1-240/Δ87-130)::PCB-All4261GAF2(C81L)His6 was purified successfully and only singly chromophorylated at All4261GAF2(C81L)His6 domain. The singly chromophorylate ApcE(1-240/Δ87-130)::PCB-All4261GAF2(C81L)His6 was incubated with fresh PCB and the doubly chromophorylated PCB-ApcE(1-240/Δ87-130)::PCB-All4261GAF2(C81L)His6 was obtained. The double chromophored fusion protein absorbed light in the range of 615–660 nm, and fluoresced only at 668 nm. Photochemistry analysis showed that excitation energy transfer from the short-wavelength absorbing at All4261GAF2(C81L) domain was achieved successfully to the long-wavelength absorbing at the ApcE(1-240/Δ87-130) domain.

Evaluating the immunostimulatory characteristics of LicKM, a novel carrier protein.

Abstract The key objective of vaccination is the induction of an effective pathogen-specific immune response that leads to protection against infection and/or disease caused by that pathogen. New vaccine technologies have resulted in second-generation recombinant vaccines containing highly purified antigens with improved tolerability and safety profiles. Unfortunately, the immune responses they induce are suboptimal without the help of adjuvants. Here, utilizing a non-toxic bacterial carrier molecule from Clostridium thermocellum a modified Lichenase (LicKM) and based upon preliminary results we hypothesize, that this serves a dual purpose; as both a mechanism to carrier dominant immunogenic epitopes for presentation to antigen presenting cells during vaccination and as an adjuvant enhancing immunity. This thermostable enzyme contains a catalytic domain loop structure separating it into two regions; an N-terminal and C-terminal region. Target protein sequences are expressed as either N or C terminal fusions; and this molecule, LicKM, may contain a single or double fusion. Currently, we are investigating the mechanism by which this molecule enhances the host humoral and cell-mediated response to vaccination. At this time, we have determined that LicKM physically binds to cells associated with the immune response. Additionally, murine vaccination in the presence of LicKM enhances antibody isotype titer production and possibly enhances the cellular responses. Our goal is to understand the immunostimulatory capability of this protein for its possible use to enhance future vaccination protocols.