Three-dimensional printing: technologies,applications, and limitations in neurosurgery

Three-dimensional (3D) printers are a developing technology penetrating a variety of markets, including the medical sector. Since its introduction to the medical field in the late 1980s, 3D printers have constructed a range of devices, such as dentures, hearing aids, and prosthetics. With the ultimate goals of decreasing healthcare costs and improving patient care and outcomes, neurosurgeons are utilizing this dynamic technology, as well. Digital Imaging and Communication in Medicine (DICOM) can be translated into Stereolithography (STL) files, which are then read and methodically built by 3D Printers. Vessels, tumors, and skulls are just a few of the anatomical structures created in a variety of materials, which enable surgeons to conduct research, educate surgeons in training, and improve pre-operative planning without risk to patients. Due to the infancy of the field and a wide range of technologies with varying advantages and disadvantages, there is currently no standard 3D printing process for patient care and medical research. In an effort to enable clinicians to optimize the use of additive manufacturing (AM) technologies, we outline the most suitable 3D printing models and computer-aided design (CAD) software for 3D printing in neurosurgery, their applications, and the limitations that need to be overcome if 3D printers are to become common practice in the neurosurgical field.     

Molecular forensics: applications,implications and limitations.

Genotyping with DNA probes can theoretically identify each person on earth. Naturally occurring variations in the nucleotide sequence of DNA (DNA sequence polymorphisms) result in genetic differences between people. The Southern blot technique can reveal characteristic DNA banding patterns at a specific genetic locus. The polymorphic DNA banding patterns at several genetic loci can be combined to help construct individual DNA "fingerprints". Such fingerprints can resolve identity in criminal and paternity cases. The appropriate technology is being used in North American law enforcement agency laboratories. Although some technical drawbacks still exist, DNA genotyping with the Southern blot technique and even newer methods will likely become the standard for individual identification. An understanding of the principles underlying DNA genotyping is required before informed decisions can be made regarding its potential widespread application.     

Phosphoproteomics by mass spectrometry: insights,implications, applications and limitations

Phosphorylation of proteins is a predominant, reversible post-translational modification. It is central to a wide variety of physiological responses and signaling mechanisms. Recent advances have allowed the global scope of phosphorylation to be addressed by mass spectrometry using phosphoproteomic approaches. In this perspective, we discuss four aspects of phosphoproteomics: the insights and implications from recently published phosphoproteomic studies and the applications and limitations of current phosphoproteomic strategies. Since approximately 50,000 known phosphorylation sites do not yet have any ascribed function, we present our perspectives on a major function of protein phosphorylation that may be of predictive value in hypothesis-based investigations. Finally, we discuss strategies to measure the stoichiometry of phosphorylation in a proteome-wide manner that is not provided by current phosphoproteomic approaches.     

Mass spectrometric analysis of neutral sphingolipids: methods, applications, and limitations

Sphingolipids represent an important class among lipids, especially when considering their vital roles in lipid metabolism. Thus, a variety of methods have been created to accomplish their analysis and the term "sphingolipidomics" has recently been coined to underline the motivation to enable a comprehensive analysis of all sphingolipid species including the acidic and the neutral ones. In this review, we summarize selected mainly biomedical based mass spectrometric approaches for the analysis of neutral sphingolipids regarding their advantages, applications and limitations. To underline some practical aspects of method development, we focus on a new method recently developed in our laboratory, which enables separation, detection, and mass spectrometric profiling of ceramide, hexosylceramide, lactosylceramide, globotriaosylceramide, globotetraosylceramide, sphingomyelin species, and cholesterol in one run. This method can be applied to investigate impairments of neutral sphingolipid metabolism in a variety of disorders such as sphingolipidoses and be employed to screen for sphingolipid profile changes as induced by knockout experiments or related studies.     

Current status of PTMs structural databases: applications,limitations and prospects

Amino Acids - Protein 3D structures, determined by their amino acid sequences, are the support of major crucial biological functions. Post-translational modifications (PTMs) play an essential role...     

Synthetic biology in various cellular and molecular fields: applications,limitations, and perspective

Molecular Biology Reports - Synthetic biology breakthroughs have facilitated genetic circuit engineering to program cells through novel biological functions, dynamic gene expressions, as well as...     

Mass spectrometry-based peptide quantification: applications and limitations

In various areas of research, proteomics and particularly the quantification of proteins and peptides renders a useful addition to biochemical experiments. The range of possible applications varies from supervision of concentration changes of relevant proteins during biogenesis to differential proteomics approaches, distinguishing, for instance, healthy and diseased states. Furthermore, mass spectrometry-based peptide quantification yields the possibility of using highly sensitive bottom-up approaches for determination of protein regulations as well as multiplexing capability. Thereby, changes in protein abundances may be linked to specific cellular states bearing the opportunity to reveal marker proteins for several diseases.     

Effects and limitations of elemental sulphur applications for enhanced phytoextraction

The application of elemental sulphur (S) to heavy metal contaminated soils is a strategy to increase metal extraction by plants. Here, we examined to which degree the efficiency of phytoextraction could be enhanced by increasing the S application rate on afield where S had already been applied for 6 years. For this purpose, the field experiment was continued for another two years doubling the S application rate on half of the S treatment plots, while continuing application at the previous rate on the other half. Doubling the application rate significantly accelerated the dissolution of calcite and the decrease in soil pH and also increased cadmium (Cd) and zinc (Zn) uptake by sunflower and tobacco. But even in a best-case-scenario remediation of the site would still take more than a century. The results indicate that we reached the maximum potential of S application to enhance metal phytoextraction on the study site. Further decrease in pH by additional S applications would bear an excessive risk of decreasing yields and increasing metal leaching out of the root zone.     

Animal models of enterovirus 71 infection: applications and limitations

Human enterovirus 71 (EV71) has emerged as a neuroinvasive virus that is responsible for several outbreaks in the Asia-Pacific region over the past 15 years. Appropriate animal models are needed to understand EV71 neuropathogenesis better and to facilitate the development of effective vaccines and drugs. Non-human primate models have been used to characterize and evaluate the neurovirulence of EV71 after the early outbreaks in late 1990s. However, these models were not suitable for assessing the neurovirulence level of the virus and were associated with ethical and economic difficulties in terms of broad application. Several strategies have been applied to develop mouse models of EV71 infection, including strategies that employ virus adaption and immunodeficient hosts. Although these mouse models do not closely mimic human disease, they have been applied to determine the pathogenesis of and treatment and prevention of the disease. EV71 receptor-transgenic mouse models have recently been developed and have significantly advanced our understanding of the biological features of the virus and the host-parasite interactions. Overall, each of these models has advantages and disadvantages, and these models are differentially suited for studies of EV71 pathogenesis and/or the pre-clinical testing of antiviral drugs and vaccines. In this paper, we review the characteristics, applications and limitation of these EV71 animal models, including non-human primate and mouse models.     

Dung fungi as a proxy for megaherbivores: opportunities and limitations for archaeological applications

The use of spores of coprophilous fungi from sedimentary sequences as proxy evidence for large herbivore abundance has garnered pronounced attention and scrutiny over the past three decades. In response to the rapid rate at which new information is being discovered on this topic, this paper presents a brief review of the archaeological applications so far, and outlines opportunities and limitations of using Sporormiella as a proxy for herbivore abundance. Specific archaeological uses of this proxy include understanding megaherbivore extinctions and human land use patterns such as pastoralism and agriculture. We analyse how dung fungal records are formed and review the mycological literature to outline factors affecting spore reproduction and preservation. These include how strongly each commonly used dung fungal taxon relies on dung as a substrate and environmental factors affecting dung fungal reproduction and coprophilous fungi deposition. Certain laboratory preparation techniques adversely affect spore representation on pollen slides. The methods of analysis and quantification of spore records also impact our understanding. We describe good practice to increase precision of analytical methods. Due to limitations imposed by some of these factors, it is possible that an absence of dung fungi from a palaeoecological record does not imply an absence of herbivores. However, consideration of these factors and inclusion of as wide a range of coprophilous spore records as possible increases the reliability of such inferences.

     

Investigating rhodamine B-labeled peptoids: scopes and limitations of its applications

The fluorophore rhodamine B is often used in biological assays. It is inexpensive, robust under a variety of reaction conditions, can be covalently linked to bioactive molecules, and has suitable spectral properties in terms of absorption and fluorescence wavelength. Nonetheless, there are some drawbacks: it can readily form a spirolactam compound, which is nonfluorescent, and therefore may not be the dye of choice for all fluorescence microscopy applications. Herein this spirolactam formation was observed by purifying such a labeled peptoid with high performance liquid chromatography (HPLC) and monitored in detail by making a series of analytical HPLC runs over time. Additionally, a small library of eight peptoids with rhodamine B as label was synthesized. Analysis of the absorption properties of these molecules demonstrated that the problem of fluorescence loss can be overcome by coupling secondary amines with rhodamine B.     

Functionalized nanocompartments (Synthosomes): limitations and prospective applications in industrial biotechnology

Synthosomes are mechanically stable vesicles with a block copolymer membrane and an engineered transmembrane protein acting as selective gate. The polymer vesicles are nanometer-sized (50-1000 nm) and functionalized by loading them with enzymes for bioconversions or encapsulating charged macromolecules for selective compound recovery/release. The Synthosome system might become a novel technology platform for biocatalysis and selective product recovery. Progress in Synthosome research comprises employed block copolymers, transmembrane channel engineering, and functionalizations, which are discussed here in detail. The challenges in transmembrane protein engineering, as well as cost-effective production, in block copolymer design and the state of the art in Synthosome characterization comprising quantification of encapsulated protein, translocation efficiency, number of transmembrane channels per vesicle, and enzyme kinetics are also presented and discussed. An assessment of the Synthosome technology platform for prospective applications in industrial (white) biotechnology concludes this review.     

Designs,applications, and limitations of genetically encoded fluorescent sensors to explore plant biology

The understanding of signaling and metabolic processes in multicellular organisms requires knowledge of the spatial dynamics of small molecules and the activities of enzymes, transporters, and other proteins in vivo, as well as biophysical parameters inside cells and across tissues. The cellular distribution of receptors, ligands, and activation state must be integrated with information about the cellular distribution of metabolites in relation to metabolic fluxes and signaling dynamics in order to achieve the promise of in vivo biochemistry. Genetically encoded sensors are engineered fluorescent proteins that have been developed for a wide range of small molecules, such as ions and metabolites, or to report biophysical processes, such as transmembrane voltage or tension. First steps have been taken to monitor the activity of transporters in vivo. Advancements in imaging technologies and specimen handling and stimulation have enabled researchers in plant sciences to implement sensor technologies in intact plants. Here, we provide a brief history of the development of genetically encoded sensors and an overview of the types of sensors available for quantifying and visualizing ion and metabolite distribution and dynamics. We further discuss the pros and cons of specific sensor designs, imaging systems, and sample manipulations, provide advice on the choice of technology, and give an outlook into future developments.

Different types of genetically encoded sensors in plants can be used to quantify and visualize ion and metabolite distributions and dynamics.     

Clinical applications of palifermin: amelioration of oral mucositis and other potential indications

Mucositis is one of the most significant toxicities in cancer patients undergoing cytotoxic treatment. It can have a negative impact on both quality of life and health economics. Severe oral mucositis can contribute to hospitalization, need for narcotic analgesics, total parentral nutrition, suboptimal delivery of anti‐neoplastic treatment, and morbidity and mortality. Palifermin, a recombinant derivative of human keratinocyte growth factor, is the first active agent approved by the FDA for the prevention of severe oral mucositis in patients undergoing haematopoietic stem cell transplantation (HSCT). Several studies have also shown significant reduction in the incidence, severity and/or duration of oral mucositis in other high‐risk settings such as concurrent chemoradiotherapy (CT/RT) for patients with head and neck cancer, and use of mucotoxic chemotherapeutic agents such as doxorubicin in sarcoma and fluorouracil for the treatment of colorectal cancer. The reduction in mucositis has translated into amelioration of symptoms and improvement in daily functioning as measured by patient‐reported outcome in multiple studies. The clinical response to palifermin appears to be related in part to epithelial proliferation and mucosal thickening. Palifermin also has other potential clinical applications including the acceleration of immune reconstitution and inhibition of graft‐versus‐host disease in patients undergoing HSCT, and mitigation of dysphagia in lung cancer patients treated with concurrent CT/RT. Palifermin is generally well tolerated with mild‐to‐moderate skin and oral adverse events. Future studies may expand the use of palifermin into other areas that would benefit from its cytoprotective and regenerative effects.