Strategies for Analysis of the Glycosylation of Proteins: Current Status and Future Perspectives

More than half of human proteins are glycosylated by a bewildering array of complex and heterogeneous N- and O-linked glycans. They function in myriad biological processes, including cell adhesion and signalling and influence the physical characteristics, stability, function, activity and immunogenicity of soluble glycoproteins. A single protein may be glycosylated differently to yield heterogenous glycoforms. Glycosylation analysis is of increasing interest in biomedical and biological research, the pharmaceutical and healthcare industry and biotechnology. This is because it is increasingly apparent that glycosylation changes in diseases, such as cancer, making it a promising target for development of clinically useful biomarkers and therapeutics. Furthermore, as the non-human cells employed in expression systems glycosylate their proteins very differently to human cells, and as glycosylation changes unpredictably under changing environmental conditions, glycans analysis for quality control, optimum efficacy and safety of recombinant glycoproteins destined for human therapeutic use is paramount. The complexities of carbohydrate chemistry make analysis challenging and while there are a variety of robust methodologies available for glycan analysis, there is currently a pressing need for the development of new, streamlined, high throughput approaches accessible to non-specialist laboratories.     

Thermophilic Hydrogen Production from Renewable Resources: Current Status and Future Perspectives

Hydrogen (H₂) is considered an alternative fuel of the future due to its high energy density and non-polluting nature. H₂ energy provides many advantages over fossil fuels in that it is renewable, eco-friendly, and efficient. The global demand for H₂ is increasing significantly; however, matching the supply of cost-competitive H₂ to meet the current demand is a major technological barrier. H₂ can be produced from lignocellulosic biomass and serve as a raw material for the synthesis of many industrially important chemicals. The use of thermophilic bacteria for biological production of H₂ appears to be a promising alternative route to the current H₂ production technologies. However, the carbon and H₂ production metabolisms in most thermophilic bacteria have not yet been completely understood. This paper summarizes the recent research progress made toward understanding the carbon utilization for H₂ production and developing gene manipulation techniques to enhance the H₂ production capabilities in thermophilic bacteria. It reviews the current status, future directions and opportunities that thermophiles can offer to enable a cost-competitive and environmentally benign H₂ production bioprocess.     

Current and Future Transgenic Whole-Cell Biosensors for Plant Macro- and Micronutrients

From the soil, plants take up macronutrients (calcium, magnesium, nitrogen, phosphorus, potassium, sulfur) and micronutrients (boron, chloride, cobalt, copper, iron, manganese, molybdenum, nickel, selenium, and zinc). In acidic soils, aluminum can interfere with nutrient uptake. There is a need for improved diagnostic tests for these soil-derived minerals that are inexpensive and sensitive, provide spatial and temporal information in plants and soil, and report bioavailable nutrient pools. A transgenic whole-cell biosensor detects a stimulus inside or outside a cell and causes a change in expression of a visible reporter such as green fluorescent protein, and thus can convert an invisible plant nutrient into a visible signal. Common transgenic whole-cell biosensors consist of promoter-reporter fusions, auxotrophs for target analytes that are transformed with constitutively expressed reporters, riboswitches and reporters based on Forster Resonance Energy Transfer (FRET). Here, we review transgenic plant biosensors that have been used to detect macronutrients and micronutrients. As plant-based biosensors are limited by the requirement to introduce and optimize a transgene in every genotype of interest, we also review microbial biosensor cells that have been used to measure plant or soil nutrients by co-inoculation with their respective extracts. Additionally, we review published transgenic whole-cell biosensors from other disciplines that have the potential to measure plant nutrients, with the goal of stimulating the development of these diagnostic technologies. We discuss current limitations and future improvements needed, and the long-term potential of transgenic whole-cell biosensors to inform plant physiology, improve soil nutrient management, and assist in breeding crops with improved nutrient use efficiency.     

Role of the 5-HT7 Receptor in the Central Nervous System: from Current Status to Future Perspectives

Pharmacological and genetic tools targeting the 5-hydroxytryptamine (5-HT)7 receptor in preclinical animal models have implicated this receptor in diverse (patho)physiological processes of the central nervous system (CNS). Some data obtained with 5-HT7 receptor knockout mice, selective antagonists, and, to a lesser extent, agonists, however, are quite contradictory. In this review, we not only discuss in detail the role of the 5-HT7 receptor in the CNS but also propose some hypothetical models, which could explain the observed inconsistencies. These models are based on two novel concepts within the field of G protein-coupled receptors (GPCR), namely biphasic signaling and G protein-independent signaling, which both have been shown to be mediated by GPCR dimerization. This led us to suggest that the 5-HT7 receptor could reside in different dimeric contexts and initiate different signaling pathways, depending on the neuronal circuitry and/or brain region. In conclusion, we highlight GPCR dimerization and G protein-independent signaling as two promising future directions in 5-HT7 receptor research, which ultimately might lead to the development of more efficient dimer- and/or pathway-specific therapeutics.     

Fish Gut Microbiome: Current Approaches and Future Perspectives

In recent years, investigations of microbial flora associated with fish gut have deepened our knowledge of the complex interactions occurring between microbes and host fish. The gut microbiome not only reinforces the digestive and immune systems in fish but is itself shaped by several host-associated factors. Unfortunately, in the past, majority of studies have focused upon the structure of fish gut microbiome providing little knowledge of effects of these factors distinctively and the immense functional potential of the gut microbiome. In this review, we have highlighted the recently gained insights into the diversity and functions of the fish gut microbiome. We have also delved on the current approaches that are being employed to study the fish gut microbiome with an aim to collate all the knowledge gained and make accurate conclusions for their application based perspectives. The literature reviewed indicated that the future research should shift towards functional microbiomics to improve the maximum sustainable yield in aquaculture.     

Glutamate Signaling in Benign and Malignant Disorders: Current Status,Future Perspectives,and Therapeutic Implications

Glutamate, a nonessential amino acid, is the major excitatory neurotransmitter in the central nervous system. As such, glutamate has been shown to play a role in not only neural processes, such as learning and memory, but also in bioenergetics, biosynthetic and metabolic oncogenic pathways. Glutamate has been the target of intense investigation for its involvement not only in the pathogenesis of benign neurodegenerative diseases (NDDs) such as Parkinson''s disease, Alzheimer''s disease, schizophrenia, multiple sclerosis, and amyotropic lateral sclerosis (ALS), but also in carcinogenesis and progression of malignant diseases. In addition to its intracellular activities, glutamate in secreted form is a phylogenetically conserved cell signaling molecule. Glutamate binding activates multiple major receptor families including the metabotropic glutamate receptors (mGluRs) and ionotropic glutamate receptors (iGluRs), both of which have been implicated in various signaling pathways in cancer. Inhibition of extracellular glutamate release or glutamate receptor activation via competitive or non-competitive antagonists decreases growth, migration and invasion and induces apoptosis in breast cancer, melanoma, glioma and prostate cancer cells. In this review, we discuss the current state of glutamate signaling research as it relates to benign and malignant diseases. In addition, we provide a synopsis of clinical trials using glutamate antagonists for the treatment of NDD and malignant diseases. We conclude that in addition to its potential role as a metabolic biomarker, glutamate receptors and glutamate-initiated signaling pathways may provide novel therapeutic opportunities for cancer.     

Clinical Use of Aromatase Inhibitors: Current Data and Future Perspectives

Abstract

A systematic procedure for the kinetic study of irreversible inhibition when the enzyme is consumed in the reaction which it catalyses, has been developed and analysed. Whereas in most reactions the enzymes are regenerated after each catalytic event and serve as reusable transacting effectors, in the consumed enzymes each catalytic center participates only once and there is no enzyme turnover. A systematic kinetic analysis of irreversible inhibition of these enzyme reactions is presented. Based on the algebraic criteria proposed in this work, it should be possible to evaluate either the mechanism of inhibition (complexing or non-complexing), or the type of inhibition (competitive, non-competitive, uncompetitive, mixed non-competitive). In addition, all kinetic constants involved in each case could be calculated. An experimental application of this analysis is also presented, concerning peptide bond formation in vitro. Using the puromycin reaction, which is a model reaction for the study of peptide bond formation in vitro and which follows the same kinetic law as the enzymes under study, we have found that: (i) the antibiotic spiramycin inhibits the puromycin reaction as a competitive irreversible inhibitor in a one step mechanism with an association rate constant equal to 1.3 × 10⁴M^-1s^-1 and, (ii) hydroxylamine inhibits the same reaction as an irreversible non-competitive inhibitor also in a one step mechanism with a rate constant equal to 1.6 × 10^-3 M^-1s^-1.     

Secondary Metabolites as Plant Traits: Current Assessment and Future Perspectives

Referee: Dr. Peter B. Kaufman, Dept. of Biology, University of Michigan, Ann Arbor, MI 48109-1048 Traditionally, secondary metabolites in plants have been investigated by phytochemists. Originally classified as waste products, these compounds more recently have been investigated extensively by ecologists and pharmacologists, and many complex biological functions have been discovered. Secondary metabolites occur nearly in all living organisms, within bacteria as well as in mammals, and are especially prominent in those organisms lacking an immune system. Functions of plant secondary metabolites comprise attractants, such as color pigments and scents, repellents such as antifeedants against insects and mammals, or toxins that affect growth and development of animal and microbial predators. Conversely, insects can employ plant-synthesized compounds to their own advantage, such as signals for feeding and oviposition and location of prey. Microbes also use secondary metabolites as carbon sources, and bacteria utilize them for quorum-sensing, an aspect recently discovered. Despite the diversity of recognized functions, the biochemical processes underlying these interactions are few. Primarily, they relate to the ability of these small molecules to bind to receptor regions of various proteins such as keys into locks. This review attempts a summary of current knowledge of secondary plant metabolism with focus on history of discovery, development of analytical techniques, theories of origin and function, signal pathways, biosynthesis, and assessment of biological activities. Outlined is current utilization by, and future perspectives in, different disciplines, such as chemosystematics, chemical ecology, and agricultural biotechnology. Examples illustrate the strong potential of research in secondary metabolism, particularly in comparison to more established disciplines such as developmental biology and physiology.     

Highly Efficient Gluten Degradation by Lactobacilli and Fungal Proteases during Food Processing: New Perspectives for Celiac Disease

Presently, the only effective treatment for celiac disease is a life-long gluten-free diet. In this work, we used a new mixture of selected sourdough lactobacilli and fungal proteases to eliminate the toxicity of wheat flour during long-time fermentation. Immunological (R5 antibody-based sandwich and competitive enzyme-linked immunosorbent assay [ELISA] and R5 antibody-based Western blot), two-dimensional electrophoresis, and mass spectrometry (matrix-assisted laser desorption ionization-time of flight, strong-cation-exchange-liquid chromatography/capillary liquid chromatography-electrospray ionization-quadrupole-time of flight [SCX-LC/CapLC-ESI-Q-TOF], and high-pressure liquid chromatography-electrospray ionization-ion trap mass spectrometry) analyses were used to determine the gluten concentration. Assays based on the proliferation of peripheral blood mononuclear cells (PBMCs) and gamma interferon production by PBMCs and intestinal T-cell lines (iTCLs) from 12 celiac disease patients were used to determine the protein toxicity of the pepsin-trypsin digests from fermented wheat dough (sourdough). As determined by R5-based sandwich and competitive ELISAs, the residual concentration of gluten in sourdough was 12 ppm. Albumins, globulins, and gliadins were completely hydrolyzed, while ca. 20% of glutenins persisted. Low-molecular-weight epitopes were not detectable by SCX-LC/CapLC-ESI-Q-TOF mass spectrometry and R5-based Western blot analyses. The kinetics of the hydrolysis of the 33-mer by lactobacilli were highly efficient. All proteins extracted from sourdough activated PBMCs and induced gamma interferon production at levels comparable to the negative control. None of the iTCLs demonstrated immunoreactivity towards pepsin-trypsin digests. Bread making was standardized to show the suitability of the detoxified wheat flour. Food processing by selected sourdough lactobacilli and fungal proteases may be considered an efficient approach to eliminate gluten toxicity.     

Lateral Flow Assays for the Diagnosis of Invasive Aspergillosis: Current Status

Purpose of Review

Diagnosis during early stages of invasive aspergillosis (IA) and targeted antifungal treatment has the potential to improve survival significantly. Despite advances in the diagnostic arsenal, invasive mold infections remain difficult to diagnose—especially at early stages before typical radiological signs develop. Varying availability and time-to-results are important limitations of current approved biomarkers and molecular assays for diagnosis of IA. Here, we will give an update on the Aspergillus-specific lateral-flow device (LFD) test. We further review promising findings on feasibility of point-of-care (POC) detection of urinary excreted fungal galactomannan-like antigens.

Recent Findings

POC LFD assays for detection of Aspergillus antigens are currently in development. The Aspergillus-specific LFD test, which is based on the JF5 antibody (Ab), detects an extracellular glycoprotein antigen secreted during active growth of Aspergillus spp. The test has shown promising results in various studies. In addition, a monoclonal Ab476-based LFD for POC detection of urinary excreted fungal galactomannan-like antigens has been developed but needs further validation.

Summary

Important advances have been made in the development of LFD assays for IA. Most promising is the Aspergillus-specific LFD test; commercial availability is still pending, however. The search for reliable POC tests for other molds, including mucorales, continues.

     

Current Status and Perspectives of Cysticercosis and Taeniasis in Japan

This mini-review describes recent epidemiological trends in cysticercosis and taeniasis in Japan. Some of the topics discussed herein were presented at the first symposium on "Current perspectives of Taenia asiatica researches", that was held in Osong in Chungbuk Province, South Korea, in October 2011 and organized by Prof. K. S. Eom, Chungbuk National University School of Medicine. To better understand the trends in the occurrence of cysticercosis and taeniasis in Japan, clinical cases reported in 2005 have been updated. In addition, the current status of Taenia asiatica infections successively occurring in Japan since 2010 is also discussed.     

应激与颞下颌关节紊乱病研究新进展

颞下颌关节紊乱病是慢性面部疼痛最常见的诱因,常常与躯体和心理主诉症状密切联系,包括疲劳,睡眠失调,焦虑和抑郁等。即使未发现任何能够合理解释疼痛原因的时候,健康专业人士也常常忽略疼痛感受的主观性。从严格的生物医学角度来讲,对疼痛的这种理解是不科学的。本文的主要目的是通过查阅近年来大量的研究文献资料,发现应激引起疼痛感觉的生物学途径以及导致颞下颌关节紊乱的原因。研究发现下丘脑-垂体-肾上腺轴、5-羟色胺和阿片样物质通路都与颌面部疼痛的发病密切相关,同时也提出了未来可能使用的治疗方法。同时,也希望本文能把与疼痛学科差别较大的口腔医学融入到需要多学科合作的颞下颌关节紊乱的诊断和治疗中,从科学角度提高对该病的临床诊疗效率。     

生物制氢的现状与发展趋势

氢能是一种理想的能源。生物制氢技术在氢能的研究和开发中占着非常重要的位置。该文介绍了生物制氢的方法和机理,综述了国内外生物制氢的现状和发展趋势并提出了作者的看法。     

Current Status and Future Strategies for Development of Transgenic Plants in China

In this review,the author summarized the current status,challenges,and strategies in China in the development oftransgenic plants and its commercialization,Based on sets of successful examples and data achieved from execution ofthe National Special Project for Transgenic Plant Research and Commercialization in the last five years,the priorities andkey directions were put forward for the future development of transgenic plants in China.