Streamlined Procedures Research Articles

Advanced structural health monitoring and damage detection using inverse methods and perturbation stiff-ness matrix analysis

Sensor technology advancements have the health monitoring task for modern intricate structures introduce many challenges, primarily centered around the augmentation of safety margins and operational dependabil-ity. The demand for diverse systems or methodologies is, therefore, increasing. These systems or methodolo-gies should be capable of detecting damage, monitoring remotely, and conducting continuous evaluations in real-time. As part of this study, inverse techniques rooted in wave propagation analysis within structural frameworks are utilized to identify damage response vectors. Additionally, mathematical formulations grounded in minimal rank perturbation theory facilitate determining a perturbed stiffness matrix for a com-promised structure. Various excitations are included in the analysis, which is based on intricate non-linear structural modeling. Using streamlined procedures, these findings confirm the inherent versatility of a range of methodologies capable of meeting both rapid monitoring needs and exhaustive analytical needs. In addition, the proposed approach promotes the use of intelligent structures embedded with a variety of sensors that are not restricted by limitations related to sensor type or spatial deployment. Monitoring structural health and detecting damage could be significantly improved by this strategic deployment.

Using a Streamlined Procedure to Combine AlCl3 and Chloroacetamide to Create a New Ionic Liquid

By reacting AlCl3 with chloroacetamide (CA) at the optimal mole ratio (2:1), a new type of ionic liquid was created in addition to the group of ionic liquids (aluminum chloride-amide). With an ionic conductivity of 0.34 mS/cm, the resultant clear brown ionic liquid demonstrated good thermal stability at ambient temperature in a glove box filled with nitrogen gas. Its coordination was established using FT-IR. Measurements have been made of additional physical characteristics including viscosity and cyclic voltammetry.

Streamlined Methods for Processing and Cryopreservation of Cell Therapy Products Using Automated Systems.

Streamlined procedures for processing and cryopreservation of cell therapies using good laboratory practices are integral to biomanufacturing process development and clinical applications. The protocol herein begins with the preparation of human cell types cultured as adherent (i.e., mesenchymal stromal cells, MSCs) or suspension cells (i.e., peripheral blood mononuclear cells, PBMCs) to comprehensively demonstrate procedures that are applicable to commonly used primary cell cultures. Cell processing steps consist of preparing high yields of cells for cryopreservation using instruments routinely used in cell manufacturing, including the Finia® Fill and Finish System and a controlled-rate freezer. The final steps comprise the storage of cells at subzero temperatures in liquid nitrogen vapor phase followed by the analysis of cell phenotypes before and after processing and cryopreservation, along with cell quality metrics for validation. Additionally, the protocol includes important considerations for the implementation of quality control measures for equipment operation and cell handling, as well as Good Laboratory Practices for cell manufacturing, which are essential for the translational use of cell therapies. Key features • The protocol applies to small- or large-scale manufacturing of cell therapy products. • It includes streamlined procedures for processing and cryopreservation of cells cultured as adherent cells (MSCs) and suspension cells (PBMCs). • Provides temperature control and rapid partitioning of sample in cryopreservation solution to maintain high viability of a range of cell types throughout the procedures. • This protocol employs the Finia® Fill and Finish System and a controlled-rate freezer. Graphical overview.

Towards Clinical Translation of In Situ Cartilage Engineering Strategies: Optimizing the Critical Facets of a Cell-Laden Hydrogel Therapy

Background:Articular cartilage repair using implantable photocrosslinkable hydrogels laden with chondrogenic cells, represents a promising in situ cartilage engineering approach for surgical treatment. The development of a surgical procedure requires a minimal viable product optimized for the clinical scenario. In our previous work we demonstrated how gelatin based photocrosslinkable hydrogels in combination with infrapatellar derived stem cells allow the production of neocartilage in vitro. In this study, we aim to optimize the critical facets of the in situ cartilage engineering therapy: the cell source, the cell isolation methodology, the cell expansion protocol, the cell number, and the delivery approach.Methods:We evaluated the impact of the critical facets of the cell-laden hydrogel therapy in vitro to define an optimized protocol that was then used in a rabbit model of cartilage repair. We performed cells counting and immunophenotype analyses, chondrogenic potential evaluation via immunostaining and gene expression, extrusion test analysis of the photocrosslinkable hydrogel, and clinical assessment of cartilage repair using macroscopic and microscopic scores.Results:We identified the adipose derived stem cells as the most chondrogenic cells source within the knee joint. We then devised a minimally manipulated stem cell isolation procedure that allows a chondrogenic population to be obtained in only 85 minutes. We found that cell expansion prior to chondrogenesis can be reduced to 5 days after the isolation procedure. We characterized that at least 5 million of cells/ml is needed in the photocrosslinkable hydrogel to successfully trigger the production of neocartilage. The maximum repairable defect was calculated based on the correlation between the number of cells retrievable with the rapid isolation followed by 5-day non-passaged expansion phase, and the minimum chondrogenic concentration in photocrosslinkable hydrogel. We next optimized the delivery parameters of the cell-laden hydrogel therapy. Finally, using the optimized procedure for in situ tissue engineering, we scored superior cartilage repair when compared to the gold standard microfracture approach.CONCLUSION:This study demonstrates the possibility to repair a critical size articular cartilage defect by means of a surgical streamlined procedure with optimized conditions.

Evaluation of dry DURAClone antibody panels for the characterization of human PBMCs, enriched T cell fractions and anti-BCMA CAR-T cells

Abstract CAR-T cells are becoming a more common therapeutic approach and are setting forth to targeting of hematological disorders other than CD19+ entities and also solid tumors. However, the immunological determinants of CAR-T efficacy still need comprehensive investigation from starting material to final product, covering different CAR constructs, transfection methods and targets. DURAClone dry antibody panels were designed to overcome standardization issues and operator-dependent impact on accuracy and precision in multicentric flow cytometry studies. The aim of this study was to evaluate the utility of DURAClone dry antibody panels for the characterization of PBMCs, derived T cell-enriched fractions as well as anti-BCMA CAR-T cells (ProMab Inc.), including MOCK-transfected controls. DURAClone dry antibody panels were used as pre-configured or extended with CAR-specific probes and further antibodies. Viable CD45+ leukocyte and CD3+ T cell count, leukocyte composition (CD3/CD19/CD56/CD14/CD16) and the T cell (CD3/CD4/CD8) maturation profile (CD45RA, CD197/CCR7, CD27, CD57) including expression of checkpoint markers (LAG-3, PD-1) were assessed. Regardless of sample type, absolute counts and cellular composition were assessed using streamlined procedures. Typical staining patterns for T cell maturation profile were observed, revealing unchanged proportions for the different stages between PBMCs and enriched T cell fractions and strong skewing toward effector cell phenotypes with elevated expression of a checkpoint markers in case of CAR-T cells. Our experiments demonstrate the flexible utility of DURAClone dry antibody panels for the phenotypic characterization from starting material to final CAR-T cell product.

COVID-19 serological survey using micro blood sampling

During August 2020, we carried out a serological survey among students and employees at the Okinawa Institute of Science and Technology Graduate University (OIST), Japan, testing for the presence of antibodies against SARS-CoV-2, the causative agent of COVID-19. We used a FDA-authorized 2-step ELISA protocol in combination with at-home self-collection of blood samples using a custom low-cost finger prick-based capillary blood collection kit. Although our survey did not find any COVID-19 seropositive individuals among the OIST cohort, it reliably detected all positive control samples obtained from a local hospital and excluded all negatives controls. We found that high serum antibody titers can persist for more than 9 months post infection. Among our controls, we found strong cross-reactivity of antibodies in samples from a serum pool from two MERS patients in the anti-SARS-CoV-2-S ELISA. Here we show that a centralized ELISA in combination with patient-based capillary blood collection using as little as one drop of blood can reliably assess the seroprevalence among communities. Anonymous sample tracking and an integrated website created a stream-lined procedure. Major parts of the workflow were automated on a liquid handler, demonstrating scalability. We anticipate this concept to serve as a prototype for reliable serological testing among larger populations.

Analyzing Mass Spectrometry Imaging Data of 13C-Labeled Phospholipids in Camelina sativa and Thlaspi arvense (Pennycress) Embryos.

The combination of 13C-isotopic labeling and mass spectrometry imaging (MSI) offers an approach to analyze metabolic flux in situ. However, combining isotopic labeling and MSI presents technical challenges ranging from sample preparation, label incorporation, data collection, and analysis. Isotopic labeling and MSI individually create large, complex data sets, and this is compounded when both methods are combined. Therefore, analyzing isotopically labeled MSI data requires streamlined procedures to support biologically meaningful interpretations. Using currently available software and techniques, here we describe a workflow to analyze 13C-labeled isotopologues of the membrane lipid and storage oil lipid intermediate―phosphatidylcholine (PC). Our results with embryos of the oilseed crops, Camelina sativa and Thlaspi arvense (pennycress), demonstrated greater 13C-isotopic labeling in the cotyledons of developing embryos compared with the embryonic axis. Greater isotopic enrichment in PC molecular species with more saturated and longer chain fatty acids suggest different flux patterns related to fatty acid desaturation and elongation pathways. The ability to evaluate MSI data of isotopically labeled plant embryos will facilitate the potential to investigate spatial aspects of metabolic flux in situ.

Design optimization and development of an Automated Storage and Retrieval System

This study aims about the planning and building up an framework for the automated material handling system by diminishing the lead time in the transportation or development of the item and furthermore concentrating on the development of the Automated Storage and Retrieval System in the businesses for expanding generation rate and furthermore to improve the material handling in warehouse. The objective of the study is to structure and manufacture an Automated Storage and Retrieval system for a mechanical stockroom and upgrading, reworking it dependent on the parameters like deals point, item prerequisite pitch or physical parameters like tallness, width, weight and so forth by utilizing a streamlining procedures.

Investigating the relationship between job satisfaction and academic brain drain: the Italian case

This work aims to show whether working in a good organisational atmosphere (job satisfaction) in the academic sector influences the propensity to migrate. This theory was tested using two surveys administered to Italian researchers abroad (IRA) and Italian researchers in Italy (IRI). The main purposes were: first, to study the relation among job satisfaction, quality of life and the propensity to migrate between IRA and IRI; second, to quantify how job satisfaction influences the academic brain drain propensity and the quality of life. Our research reveals a dual state of Italian academic job satisfaction. Those researchers who live abroad report satisfaction for all the four dimensions of job satisfaction, which influences positively their quality of life and decreases their propensity to return to Italy. In contrast, researchers that remain in Italy adapt to the system and record negative perception of job satisfaction, low quality of life and show propensity to emigrate abroad. The Italian academic environment must improve its status with an emphasis on organisational and job satisfaction. This can be done through a direct intervention in organisational administration favouring streamlined procedures, functional research productivity, harmony of academic standards and a meritocratic reward process.

Anesthesia for the patient undergoing total knee replacement: current status and future prospects.

Total knee arthroplasty (TKA) has become one of the most common orthopedic surgical procedures performed nationally. As the population and surgical techniques for TKAs have evolved over time, so have the anesthesia and analgesia used for these procedures. General anesthesia has been the dominant form of anesthesia utilized for TKA in the past, but regional anesthetic techniques are on the rise. Multiple studies have shown the potential for regional anesthesia to improve patient outcomes, such as a decrease in intraoperative blood loss, length of stay, and patient mortality. Anesthesiologists are also moving toward multimodal analgesia, which includes peripheral nerve blockade, periarticular injection, and preemptive analgesia. The goal of multimodal analgesia is to improve perioperative pain control while minimizing systemic narcotic consumption. With improved postoperative pain management and rapid patient rehabilitation, new clinical pathways have been engineered to fast track patient recovery after orthopedic procedures. The aim of these clinical pathways was to improve quality of care, minimize unnecessary variations in care, and reduce cost by using streamlined procedures and protocols. The future of TKA care will be formalized clinical pathways and tracks to better optimize perioperative algorithms with regard to pain control and perioperative rehabilitation.

Root Resorption with Orthodontic Intrusion Review

AIM AND OBJECTIVE: This article reviews root resorption with micro implant assisted intrusion BACKGROUND: Orthodontic intrusion is a common treatment approach in managing orthodontic esthetic and functional problems, including gummy smile and deep bite. Amongst, different type of orthodontic procedures, intrusion is found to be undoubtedly a difficult procedure. Micro implants are found to produce offer an effective skeletal anchorage which is an assist to orthodontist since it renders the intrusion of both anteriors and posterior teeth an increasingly streamlined procedure from mechanical support. Apical root resorption is abnormal consequences in orthodontic treatment. Root resorption produce shortening and breakdown the integrity of teeth from the arch. This article reviews the systemic evaluation of root resorption with micro implants assisted intrusion REASON: The purpose of the systemic review is evaluate the various factor of root resorption with micro implant assisted with intrusion.

Quantifying uncertainty in brain network measures using Bayesian connectomics.

The wiring diagram of the human brain can be described in terms of graph measures that characterize structural regularities. These measures require an estimate of whole-brain structural connectivity for which one may resort to deterministic or thresholded probabilistic streamlining procedures. While these procedures have provided important insights about the characteristics of human brain networks, they ultimately rely on unwarranted assumptions such as those of noise-free data or the use of an arbitrary threshold. Therefore, resulting structural connectivity estimates as well as derived graph measures fail to fully take into account the inherent uncertainty in the structural estimate. In this paper, we illustrate an easy way of obtaining posterior distributions over graph metrics using Bayesian inference. It is shown that this posterior distribution can be used to quantify uncertainty about graph-theoretical measures at the single subject level, thereby providing a more nuanced view of the graph-theoretical properties of human brain connectivity. We refer to this model-based approach to connectivity analysis as Bayesian connectomics.

Developing translational research infrastructure and capabilities associated with cancer clinical trials

The integration of molecular information in clinical decision making is becoming a reality. These changes are shaping the way clinical research is conducted, and as reality sets in, the challenges in conducting, managing and organising multi-disciplinary research become apparent. Clinical trials provide a platform to conduct translational research (TR) within the context of high quality clinical data accrual. Integrating TR objectives in trials allows the execution of pivotal studies that provide clinical evidence for biomarker-driven treatment strategies, targeting early drug development trials to a homogeneous and well defined patient population, supports the development of companion diagnostics and provides an opportunity for deepening our understanding of cancer biology and mechanisms of drug action. To achieve these goals within a clinical trial, developing translational research infrastructure and capabilities (TRIC) plays a critical catalytic role for translating preclinical data into successful clinical research and development. TRIC represents a technical platform, dedicated resources and access to expertise promoting high quality standards, logistical and operational support and unified streamlined procedures under an appropriate governance framework. TRIC promotes integration of multiple disciplines including biobanking, laboratory analysis, molecular data, informatics, statistical analysis and dissemination of results which are all required for successful TR projects and scientific progress. Such a supporting infrastructure is absolutely essential in order to promote high quality robust research, avoid duplication and coordinate resources. Lack of such infrastructure, we would argue, is one reason for the limited effect of TR in clinical practice beyond clinical trials.

Physician's assistants: a workforce solution for Australia?

Significant medical workforce shortages, particularly in rural and remote locations, have prompted a range of responses in Australia at both state and Commonwealth levels. One such response was a pilot project to test the suitability of the Physician Assistant (PA) role in the Australian context. Five US-trained and accredited PAs were employed by Queensland Health and deployed in urban, rural and remote settings across Queensland. A concurrent mixed-method evaluation was conducted by Urbis, an independent research firm. The evaluation found that the PAs provided quality, safe clinical care under the supervision of local medical officers. The majority of nurses and doctors who worked with the PAs believed that the PAs made a positive contribution to the health care team by increasing capacity to meet patient needs; reducing on-call requirements for doctors; liaising with other clinical team members; streamlining procedures for efficient patient throughput; and providing continuity during periods of doctor changeover. The Pilot demonstrated that a delegated PA role can provide safe, quality health care by augmenting an established healthcare team. The PA role has the potential to benefit the community by increasing the capacity of the health care system, and to improve recruitment and retention by providing an additional professional pathway. The small size of the Pilot limits the ability to generalise regarding the future efficacy of the PA role in Australia. Further research is required to test training and deployment of PAs in a wider range of Australian clinical settings, including general practice and rural health clinics.

Making the Cut: Options for Making Initial Evaluations of Malting Quality in Barley

Alternatives to traditional methods of estimating the potential malting quality of barley lines were examined through their application to two regional barley nurseries, with the goal of identifying streamlined procedures suitable for making initial assessments of malting quality. Mashing malt using an isothermal hot water extract (HWE) procedure instead of the standard Congress mash generated distinct but closely related values for primary measures of malt modification, including malt extract, wort soluble protein, soluble/total malt protein, wort β-glucan, and wort free amino nitrogen. Simple measurements of other wort properties, including refractive index and wort osmolyte concentration, also showed strong correlations between Congress and HWE worts. Analyses of small-scale (microcentrifuge tube) versions of isothermal HWE worts showed good correlations with the attributes of Congress worts. Although the key malting quality metrics for Congress and HWE worts were not equivalent, their close correlations suggest that they may be equally able to differentiate the malting quality of barley lines. The simple isothermal temperature profile of the HWE wort, in conjunction with implementation of methods that use simple laboratory instrumentation for wort analysis, could simplify initial malting quality analysis and make it more accessible to laboratories lacking dedicated mashing and wort analysis instruments.

Achieving Sustainable Mobility in Developing Countries: Suggestions for a Post-2012 Agreement

In December 2009, countries meet in Copenhagen to establish a new global climate agreement. This article links the need for reducing transport-related greenhouse gas emissions in developing countries with the current international climate negotiations. Arguing that a sustainable transport approach requires comprehensive policy packages, it assesses the suitability of current climate negotiation proposals in promoting sustainable transport. The project-based approach under the current climate regime incentivises neither comprehensive sustainable transport and mobility policies, nor sufficient numbers of local projects. Current proposals to increase efforts by developing countries, to reform the Clean Development Mechanism, and to create new emission trading mechanisms are promising but still have to overcome several obstacles. One obstacle involves how to properly assess the impact of actions while maintaining streamlined procedures. The authors conclude from their analysis that the best way forward would be to establish an international mitigation fund with a dedicated transport window financed by industrialised countries. This fund would enable developing countries to implement national policies and local projects. Developing countries would outline low-carbon development strategies, including a sectoral strategy for low-carbon transport.

Climate change and sustainable development

Th is paper argues that in the future the primary focus of policy research and global agreements should be the de-carbonization of economic development. Consequently, instead of treating climate stabilization and economic development as separate and equal, the strategy should be to re-integrate the two global policy goals, in part by separating responsibility (and funding) from action. Th is will require an approach that goes beyond Kyoto. Th e paper invokes the example of the Manhattan Project to argue for a massive, globally funded public investment program for the deployment of renewable energy technologies in developing countries.

SPARE: A qualitative study investigating randomization barriers in a Selective Bladder Preservation trial (SBP) (ISCRCTN: 61126465)

5077 Background: Recruitment to randomised trials is challenging, especially when treatment strategies are complex. While studies have demonstrated barriers to recruitment, few have investigated why patients accept or refuse randomisation. The SPARE feasibility study included such an investigation to highlight difficulties that might be dealt with as the study progressed and to inform procedures for a subsequent phase III trial. Methods: Patients had newly diagnosed invasive bladder cancer and were receiving neoadjuvant chemotherapy prior to invitation to a randomised trial (SPARE) that offered the possibility of radical cystectomy or SBP. Patients allocated SBP received cystectomy or radiotherapy dependent on chemotherapy response. 15 acceptors and 10 decliners to randomisation participated in the qualitative substudy. Methods included recorded transcribed interviews and a ‘framework analysis.’ Results: Both groups experienced initial confusion, ‘muddled’ communication, information overload, and lack of time leading to misunderstandings about the trial. Perceived side effects did not necessarily inform decisions. Decision to participate was seldom made unilaterally. Individual specialist input by, and continuity between, professionals and their patients, appeared to make decision making easier. Acceptors were inclined to altruism and trust in the medical profession, often agreeing to participate with minimum understanding regarding randomisation and/or equipoise and a ‘nothing to lose’ attitude as long as withdrawal from the trial was possible. Decliners tended to opt for radiotherapy, perceived as a rational choice in the context of this trial; many abhorred the thought of surgery that was perceived as undermining and a ‘treatment in reserve.’ Conclusions: Health professionals need to consider streamlining procedures and recognise patient preferences in terms of treatments offered and information received. Recognition of the complexity of communicating equipoise and randomisation may increase ‘informed consent’ levels and recruitment rates. A further qualitative substudy investigating doctor/patient consultations in real time with confidential feedback is proposed to optimise recruitment rates. No significant financial relationships to disclose.

Efficient mouse transgenesis using Gateway-compatible ROSA26 locus targeting vectors and F1 hybrid ES cells

The ability to rapidly and efficiently generate reliable Cre/loxP conditional transgenic mice would greatly complement global high-throughput gene targeting initiatives aimed at identifying gene function in the mouse. We report here the generation of Cre/loxP conditional ROSA26-targeted ES cells within 3–4 weeks by using Gateway® cloning to build the target vectors. The cDNA of the gene of interest can be expressed either directly by the ROSA26 promoter providing a moderate level of expression or by a CAGG promoter placed in the ROSA26 locus providing higher transgene expression. Utilization of F1 hybrid ES cells with exceptional developmental potential allows the production of germ line transmitting, fully or highly ES cell-derived mice by aggregation of cells with diploid embryos. The presented streamlined procedures accelerate the examination of phenotypical consequences of transgene expression. It also provides a unique tool for comparing the biological activity of polymorphic or splice variants of a gene, or products of different genes functioning in the same or parallel pathways in an overlapping manner.

Patent procedure in England and Wales: the Streamlined Procedure meets resistance

The proposition by the English High Court in 2005 that the Streamlined Procedure for dealing with patent actions should apply whenever raised by a party to an action in the absence of convincing reasons to the contrary has been rejected by the Court of Appeal.