Glycan Labeling and Analysis in Cells and In Vivo

2021 ◽  
Vol 14 (1) ◽  
pp. 363-387
Author(s):  
Bo Cheng ◽  
Qi Tang ◽  
Che Zhang ◽  
Xing Chen
Keyword(s):  
Chemical Modification ◽  
Biological Function ◽  
Biological Processes ◽  
Affinity Tags ◽  
Functional Roles ◽  
Antibody Conjugates ◽  
Living Organisms ◽  
In Cells

As one of the major types of biomacromolecules in the cell, glycans play essential functional roles in various biological processes. Compared with proteins and nucleic acids, the analysis of glycans in situ has been more challenging. Herein we review recent advances in the development of methods and strategies for labeling, imaging, and profiling of glycans in cells and in vivo. Cellular glycans can be labeled by affinity-based probes, including lectin and antibody conjugates, direct chemical modification, metabolic glycan labeling, and chemoenzymatic labeling. These methods have been applied to label glycans with fluorophores, which enables the visualization and tracking of glycans in cells, tissues, and living organisms. Alternatively, labeling glycans with affinity tags has enabled the enrichment of glycoproteins for glycoproteomic profiling. Built on the glycan labeling methods, strategies enabling cell-selective and tissue-specific glycan labeling and protein-specific glycan imaging have been developed. With these methods and strategies, researchers are now better poised than ever to dissect the biological function of glycans in physiological or pathological contexts.

scholarly journals Treating Senescence like Cancer: Novel Perspectives in Senotherapy of Chronic Diseases

2020 ◽  
Vol 21 (21) ◽  
pp. 7984
Author(s):  
Alessia Mongelli ◽  
Sandra Atlante ◽  
Veronica Barbi ◽  
Tiziana Bachetti ◽  
Fabio Martelli ◽  
...  
Keyword(s):  
Chronic Diseases ◽  
Cell Senescence ◽  
Biological Processes ◽  
Social Relevance ◽  
Chemical Agents ◽  
Age Related ◽  
Living Organisms ◽  
Specific Pathway

The WHO estimated around 41 million deaths worldwide each year for age-related non-communicable chronic diseases. Hence, developing strategies to control the accumulation of cell senescence in living organisms and the overall aging process is an urgently needed problem of social relevance. During aging, many biological processes are altered, which globally induce the dysfunction of the whole organism. Cell senescence is one of the causes of this modification. Nowadays, several drugs approved for anticancer therapy have been repurposed to treat senescence, and others are under scrutiny in vitro and in vivo to establish their senomorphic or senolytic properties. In some cases, this research led to a significant increase in cell survival or to a prolonged lifespan in animal models, at least. Senomorphics can act to interfere with a specific pathway in order to restore the appropriate cellular function, preserve viability, and to prolong the lifespan. On the other hand, senolytics induce apoptosis in senescent cells allowing the remaining non–senescent population to preserve or restore tissue function. A large number of research articles and reviews recently addressed this topic. Herein, we would like to focus attention on those chemical agents with senomorphic or senolytic properties that perspectively, according to literature, suggest a potential application as senotherapeutics for chronic diseases.

scholarly journals The Beneficial Effects of Morusin, an Isoprene Flavonoid Isolated from the Root Bark of Morus

2020 ◽  
Vol 21 (18) ◽  
pp. 6541
Author(s):  
Dong Wook Choi ◽  
Sang Woo Cho ◽  
Seok-Geun Lee ◽  
Cheol Yong Choi
Keyword(s):  
Inflammatory Diseases ◽  
Root Bark ◽  
Biological Processes ◽  
Functional Roles ◽  
Beneficial Effects ◽  
Biochemical Identification ◽  
Underlying Mechanisms ◽  
Clinical Potential

The root bark of Morus has long been appreciated as an antiphlogistic, diuretic and expectorant drug in Chinese herbal medicine, albeit with barely known targets and mechanisms of action. In the 1970s, the development of analytic chemistry allowed for the discovery of morusin as one of 7 different isoprene flavonoid derivatives in the root bark of Morus. However, the remarkable antioxidant capacity of morusin with the unexpected potential for health benefits over the other flavonoid derivatives has recently sparked scientific interest in the biochemical identification of target proteins and signaling pathways and further clinical relevance. In this review, we discuss recent advances in the understanding of the functional roles of morusin in multiple biological processes such as inflammation, apoptosis, metabolism and autophagy. We also highlight recent in vivo and in vitro evidence on the clinical potential of morusin treatment for multiple human pathologies including inflammatory diseases, neurological disorders, diabetes, cancer and the underlying mechanisms.

scholarly journals IL7R⍺ is required for hematopoietic stem cell reconstitution of tissue-resident lymphoid cells

2021 ◽  
Author(s):  
Atesh K Worthington ◽  
Taylor S Cool ◽  
Donna M Poscablo ◽  
Adeel Hussaini ◽  
Anna E Beaudin ◽  
...  
Keyword(s):  
Cell Types ◽  
Lineage Tracing ◽  
Lymphoid Cells ◽  
Lymphoid Tissues ◽  
Hematopoietic Stem ◽  
Functional Roles ◽  
Reciprocal Transplants ◽  
B1a Cells

Traditional, adult-derived lymphocytes that circulate provide adaptive immunity to infection and pathogens. However, subsets of lymphoid cells are also found in non-lymphoid tissues and are called tissue-resident lymphoid cells (TLCs). TLCs encompass a wide array of cell types that span the spectrum of innate-to-adaptive immune function. Unlike traditional lymphocytes that are continuously generated from hematopoietic stem cells (HSCs), many TLCs are of fetal origin and poorly generated from adult HSCs. Here, we sought to understand the development of murine TLCs across multiple tissues and therefore probed the roles of Flk2 and IL7R⍺, two cytokine receptors with known roles in traditional lymphopoiesis. Using Flk2- and Il7r-Cre lineage tracing models, we found that peritoneal B1a cells, splenic marginal zone B (MZB) cells, lung ILC2s and regulatory T cells (Tregs) were highly labeled in both models. Despite this high labeling, highly quantitative, in vivo functional approaches showed that the loss of Flk2 minimally affected the generation of these cells in situ. In contrast, the loss of IL7R⍺, or combined deletion of Flk2 and IL7R⍺, dramatically reduced the cell numbers of B1a cells, MZBs, ILC2s, and Tregs both in situ and upon transplantation, indicating an intrinsic and more essential role for IL7Rα. Surprisingly, reciprocal transplants of WT HSCs showed that an IL7Rα-/- environment selectively impaired reconstitution of TLCs when compared to TLC numbers in situ. Taken together, our data revealed functional roles of Flk2 and IL7Rα in the establishment of tissue-resident lymphoid cells.

scholarly journals Phosphorothioate Antisense Oligonucleotides Induce the Formation of Nuclear Bodies

1998 ◽  
Vol 9 (5) ◽  
pp. 1007-1023 ◽  
Author(s):  
Peter Lorenz ◽  
Brenda F. Baker ◽  
C. Frank Bennett ◽  
David L. Spector
Keyword(s):  
Antisense Oligonucleotides ◽  
De Novo ◽  
Regulation Of Gene Expression ◽  
Nuclear Body ◽  
Nuclear Bodies ◽  
Phosphorothioate Oligodeoxynucleotides ◽  
Nuclear Structures ◽  
In Cells

Antisense oligonucleotides are powerful tools for the in vivo regulation of gene expression. We have characterized the intracellular distribution of fluorescently tagged phosphorothioate oligodeoxynucleotides (PS-ONs) at high resolution under conditions in which PS-ONs have the potential to display antisense activity. Under these conditions PS-ONs predominantly localized to the cell nucleus where they accumulated in 20–30 bright spherical foci designated phosphorothioate bodies (PS bodies), which were set against a diffuse nucleoplasmic population excluding nucleoli. PS bodies are nuclear structures that formed in cells after PS-ON delivery by transfection agents or microinjection but were observed irrespectively of antisense activity or sequence. Ultrastructurally, PS bodies corresponded to electron-dense structures of 150–300 nm diameter and resembled nuclear bodies that were found with lower frequency in cells lacking PS-ONs. The environment of a living cell was required for the de novo formation of PS bodies, which occurred within minutes after the introduction of PS-ONs. PS bodies were stable entities that underwent noticeable reorganization only during mitosis. Upon exit from mitosis, PS bodies were assembled de novo from diffuse PS-ON pools in the daughter nuclei. In situ fractionation demonstrated an association of PS-ONs with the nuclear matrix. Taken together, our data provide evidence for the formation of a nuclear body in cells after introduction of phosphorothioate oligodeoxynucleotides.

scholarly journals Iodine: Its Role in Thyroid Hormone Biosynthesis and Beyond

Nutrients ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 4469
Author(s):  
Salvatore Sorrenti ◽  
Enke Baldini ◽  
Daniele Pironi ◽  
Augusto Lauro ◽  
Valerio D'Orazi ◽  
...  
Keyword(s):  
Cancer Progression ◽  
Biological Function ◽  
Human Cancer ◽  
Oxygen Species ◽  
Human Cancer Cell Lines ◽  
Functional Roles ◽  
Thyroid Hormone Biosynthesis ◽  
Evolutionarily Conserved ◽  
Hormone Biosynthesis ◽  
Living Organisms

The present review deals with the functional roles of iodine and its metabolism. The main biological function of iodine concerns its role in the biosynthesis of thyroid hormones (THs) by the thyroid gland. In addition, however, further biological roles of iodine have emerged. Precisely, due to its significant action as scavenger of reactive oxygen species (ROS), iodine is thought to represent one of the oldest antioxidants in living organisms. Moreover, iodine oxidation to hypoiodite (IO−) has been shown to possess strong bactericidal as well as antiviral and antifungal activity. Finally, and importantly, iodine has been demonstrated to exert antineoplastic effects in human cancer cell lines. Thus, iodine, through the action of different tissue-specific peroxidases, may serve different evolutionarily conserved physiological functions that, beyond TH biosynthesis, encompass antioxidant activity and defense against pathogens and cancer progression.

5-lipoxygenase expression in dendritic cells generated from CD34+ hematopoietic progenitors and in lymphoid organs

Blood ◽  
2000 ◽  
Vol 96 (12) ◽  
pp. 3857-3865
Author(s):  
Rainer Spanbroek ◽  
Markus Hildner ◽  
Dieter Steinhilber ◽  
Norbert Fusenig ◽  
Kozo Yoneda ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
T Lymphocyte ◽  
Transforming Growth Factor ◽  
Leukotriene B4 ◽  
Lymphoid Tissues ◽  
Ionophore A23187 ◽  
In Cells

The 5-lipoxygenase (5-LO) pathway in human CD34+ hematopoietic progenitor cells, which were induced to differentiate into dendritic cells (DCs) by cytokines in vitro and in DCs of lymphoid tissues in situ, was examined. Extracts prepared from HPCs contained low levels of 5-LO or 5-LO–activating protein. Granulocyte-macrophage colony-stimulating factor (GM-CSF) plus tumor necrosis factor–α (TNF-α) promoted DC differentiation and induced a strong rise in 5-LO and FLAP expression. Fluorescence-activated cell sorter (FACS) analyses identified a major DC population coexpressing human leukocyte antigen (HLA)-DR/CD80 and monocytic or Langerhans cell markers. Transforming growth factor–β1 (TGF-β–1), added to support DC maturation, strongly promoted the appearance of CD1a+/Lag+ Langerhans-type cells as well as mature CD83+ DCs. TGF-β–1 further increased 5-LO and FLAP expression, recruited additional cells into the 5-LO+DC population, and promoted production of 5-hydroxyeicosatetraenoic acid and leukotriene B4 in response to calcium (Ca++) ionophore A23187. These in vitro findings were corroborated by 5-LO expression in distinct DC phenotypes in vivo. Scattered 5-LO and FLAP in situ hybridization signals were recorded in cells of paracortical T-lymphocyte–rich areas and germinal centers (GCs) of lymph nodes (LNs) and tonsil and in cells of mucosae overlying the Waldeyer tonsillar ring. 5-LO protein localized to both CD1a+ immature DCs and to CD83+ mature interdigitating DCs of T-lymphocyte–rich areas of LNs and tonsil. As DCs have the unique ability to initiate naive lymphocyte activation, our data support the hypothesis that leukotrienes act at proximal steps of adaptive immune responses.

scholarly journals Advancements in Non-Invasive Biological Surface Sampling and Emerging Applications

Separations ◽  
2019 ◽  
Vol 6 (4) ◽  
pp. 52 ◽  
Author(s):  
Atakan Arda Nalbant ◽  
Ezel Boyacı
Keyword(s):  
Ocular Surface ◽  
Passive Samplers ◽  
Tissue Fluid ◽  
Surface Sampling ◽  
Future Technology ◽  
Non Invasive ◽  
Biological Surfaces ◽  
Living Organisms

Biological surfaces such as skin and ocular surface provide a plethora of information about the underlying biological activity of living organisms. However, they pose unique problems arising from their innate complexity, constant exposure of the surface to the surrounding elements, and the general requirement of any sampling method to be as minimally invasive as possible. Therefore, it is challenging but also rewarding to develop novel analytical tools that are suitable for in vivo and in situ sampling from biological surfaces. In this context, wearable extraction devices including passive samplers, extractive patches, and different microextraction technologies come forward as versatile, low-invasive, fast, and reliable sampling and sample preparation tools that are applicable for in vivo and in situ sampling. This review aims to address recent developments in non-invasive in vivo and in situ sampling methods from biological surfaces that introduce new ways and improve upon existing ones. Directions for the development of future technology and potential areas of applications such as clinical, bioanalytical, and doping analyses will also be discussed. These advancements include various types of passive samplers, hydrogels, and polydimethylsiloxane (PDMS) patches/microarrays, and other wearable extraction devices used mainly in skin sampling, among other novel techniques developed for ocular surface and oral tissue/fluid sampling.

Nanoporous Gold: A Biomaterial for Microfabricated Drug-Delivery Platforms

2012 ◽  
Vol 1415 ◽  
Author(s):  
Erkin Seker ◽  
Yevgeny Berdichevsky ◽  
Kevin J. Staley ◽  
Martin L. Yarmush
Keyword(s):  
Drug Delivery ◽  
Electrochemical Sensors ◽  
Volume Ratio ◽  
Nanostructured Material ◽  
Nanoporous Gold ◽  
Biological Processes ◽  
Au Thin Films

ABSTRACTNanoporous gold (np-Au) is a promising nanostructured material with many desirable properties, including large surface area-to-volume ratio, corrosion resistance, high conductivity, and well-studied thiol-based surface chemistry. While np-Au has been used in a variety of applications, from fuel cells to electrochemical sensors, its interface with biology, where many of its exciting applications lie, is surprisingly non-existent. This paper reports on drug delivery from np-Au thin films for modifying cell proliferation in situ. We expect that establishing np-Au as a biomaterial with drug delivery capabilities will create new opportunities for engineering advanced BioMEMS devices that can monitor and modulate biological processes in both in vitro and in vivo settings.

scholarly journals 2′O-Ribose Methylation of Ribosomal RNAs: Natural Diversity in Living Organisms, Biological Processes, and Diseases

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1948
Author(s):  
Mariam Jaafar ◽  
Hermes Paraqindes ◽  
Mathieu Gabut ◽  
Jean-Jacques Diaz ◽  
Virginie Marcel ◽  
...  
Keyword(s):  
Chemical Modification ◽  
Single Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Human Cell ◽  
Biological Processes ◽  
Human Cell Lines ◽  
Natural Diversity ◽  
Living Organisms ◽  
Line Organ

Recent findings suggest that ribosomes, the translational machineries, can display a distinct composition depending on physio-pathological contexts. Thanks to outstanding technological breakthroughs, many studies have reported that variations of rRNA modifications, and more particularly the most abundant rRNA chemical modification, the rRNA 2′O-ribose methylation (2′Ome), intrinsically occur in many organisms. In the last 5 years, accumulating reports have illustrated that rRNA 2′Ome varies in human cell lines but also in living organisms (yeast, plant, zebrafish, mouse, human) during development and diseases. These rRNA 2′Ome variations occur either within a single cell line, organ, or patient’s sample (i.e., intra-variability) or between at least two biological conditions (i.e., inter-variability). Thus, the ribosomes can tolerate the absence of 2′Ome at some specific positions. These observations question whether variations in rRNA 2′Ome could provide ribosomes with particular translational regulatory activities and functional specializations. Here, we compile recent studies supporting the heterogeneity of ribosome composition at rRNA 2′Ome level and provide an overview of the natural diversity in rRNA 2′Ome that has been reported up to now throughout the kingdom of life. Moreover, we discuss the little evidence that suggests that variations of rRNA 2′Ome can effectively impact the ribosome activity and contribute to the etiology of some human diseases.

scholarly journals Molecular characterization and functional analysis of the lysosomal cathepsin D-like gene in red swamp crayfish, Procambarus clarkii

Genome ◽  
2021 ◽  
pp. 1-11
Author(s):  
Long Huang ◽  
Ben-li Wu ◽  
Ji-Xiang He ◽  
Ye Zhang ◽  
Jing Chen ◽  
...  
Keyword(s):  
Cathepsin D ◽  
Procambarus Clarkii ◽  
Distribution Analysis ◽  
Aspartic Proteinases ◽  
Reading Frame ◽  
Functional Roles ◽  
Polycytidylic Acid ◽  
Red Swamp Crayfish ◽  
Living Organisms

Aspartic proteinases are one of the four families of proteinase enzymes that are widely present in living organisms. They are involved in various physiological events, such as protein degradation, development, and host defense. However, the characterization and functional roles of aspartic proteinases remain to be elucidated in crustaceans. Here, we characterized a fragment of cathepsin D-like cDNA from red swamp crayfish, Procambarus clarkii (Pc-cathepsin D-like). The open reading frame of the Pc-cathepsin D-like gene contained 1152 bp, encoding a protein of 383 amino acid residues. We also evaluated the immunological role of the Pc-cathepsin D-like gene in vivo. Spatial distribution analysis revealed that the Pc-cathepsin D-like mRNA was high in the hepatopancreas, followed by the gut, gills, and hemocytes of P. clarkii. The expression levels of the Pc-cathepsin D-like gene increased following challenge with viral (polyinosinic: polycytidylic acid) and bacterial (lipopolysaccharides, peptidoglycan) PAMPs compared with PBS injection. The suppression of the Pc-cathepsin D-like gene by RNA interference significantly increased the expression of immune-associated genes. These results showed that the Pc-cathepsin D-like gene has an essential biological role in innate immune responses because it regulates the expression of immune-associated genes.

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