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  1. Article ; Online: Single-Domain Antibodies for Intracellular Toxin Neutralization.

    Czajka, Timothy F / Mantis, Nicholas J

    Methods in molecular biology (Clifton, N.J.)

    2022  Volume 2446, Page(s) 469–487

    Abstract: Ricin is a plant-derived toxin with a history as a biothreat agent. The toxin's enzymatic subunit, ricin toxin A chain (RTA), is a ribosome-inactivating protein that, when delivered into the cytoplasm of mammalian cells, arrests protein synthesis with ... ...

    Abstract Ricin is a plant-derived toxin with a history as a biothreat agent. The toxin's enzymatic subunit, ricin toxin A chain (RTA), is a ribosome-inactivating protein that, when delivered into the cytoplasm of mammalian cells, arrests protein synthesis with extraordinary efficiency. Once within the cytoplasm, RTA is shielded from circulating toxin-neutralizing antibodies. Here, we describe methods we developed to neutralize RTA within the cytoplasm of Vero cells using DNA-based delivery of alpaca-derived single-domain antibodies (VHHs) targeting RTA's active site. We describe the design of the VHH expression vectors, assessment of transient expression of VHHs in Vero cells by enzyme-linked immunosorbent assay and western blotting, and cytotoxicity studies. While the protocols here are specific to ricin, they are easily modified for other toxins or even intracellular pathogens such as viruses.
    MeSH term(s) Animals ; Antibodies, Neutralizing ; Camelids, New World ; Chlorocebus aethiops ; Ricin ; Single-Domain Antibodies/genetics ; Vero Cells
    Chemical Substances Antibodies, Neutralizing ; Single-Domain Antibodies ; Ricin (9009-86-3)
    Language English
    Publishing date 2022-02-14
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-2075-5_24
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Slaying SARS-CoV-2 One (Single-domain) Antibody at a Time.

    Czajka, Timothy F / Vance, David J / Mantis, Nicholas J

    Trends in microbiology

    2020  Volume 29, Issue 3, Page(s) 195–203

    Abstract: Camelid-derived and synthetic single-domain antibodies (sdAbs) are emerging as potent weapons against the novel coronavirus, SARS-CoV-2. sdAbs are small, compact, thermostable immunoglobulin elements capable of binding targets with subnanomolar ... ...

    Abstract Camelid-derived and synthetic single-domain antibodies (sdAbs) are emerging as potent weapons against the novel coronavirus, SARS-CoV-2. sdAbs are small, compact, thermostable immunoglobulin elements capable of binding targets with subnanomolar affinities. By leveraging the power of phage- and yeast surface-display technologies, rare sdAbs can be isolated from highly diverse and complex antibody libraries. Once in hand, sdAbs can be engineered to improve binding affinity, avidity, target specificities, and biodistribution. In this Opinion piece we highlight a series of sophisticated studies describing the identification of ultrapotent sdAbs directed against the receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein. We discuss the possible applications of these antibodies in the global fight against COVID-19.
    MeSH term(s) Antibodies, Viral/chemistry ; Antibodies, Viral/immunology ; Humans ; Protein Structure, Tertiary ; Single-Domain Antibodies/chemistry ; Single-Domain Antibodies/immunology ; Spike Glycoprotein, Coronavirus/immunology ; Synthetic Biology/methods
    Chemical Substances Antibodies, Viral ; Single-Domain Antibodies ; Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2
    Language English
    Publishing date 2020-12-16
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Review
    ZDB-ID 1158963-2
    ISSN 1878-4380 ; 0966-842X
    ISSN (online) 1878-4380
    ISSN 0966-842X
    DOI 10.1016/j.tim.2020.12.006
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 3738: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (2.OG)
    ab Jg. 2022: Lesesaal (EG)

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  3. Article ; Online: Single-domain antibodies neutralize ricin toxin intracellularly by blocking access to ribosomal P-stalk proteins.

    Czajka, Timothy F / Vance, David J / Davis, Simon / Rudolph, Michael J / Mantis, Nicholas J

    The Journal of biological chemistry

    2022  Volume 298, Issue 4, Page(s) 101742

    Abstract: During ricin intoxication in mammalian cells, ricin's enzymatic (RTA) and binding (RTB) subunits disassociate in the endoplasmic reticulum. RTA is then translocated into the cytoplasm where, by virtue of its ability to depurinate a conserved residue ... ...

    Abstract During ricin intoxication in mammalian cells, ricin's enzymatic (RTA) and binding (RTB) subunits disassociate in the endoplasmic reticulum. RTA is then translocated into the cytoplasm where, by virtue of its ability to depurinate a conserved residue within the sarcin-ricin loop (SRL) of 28S rRNA, it functions as a ribosome-inactivating protein. It has been proposed that recruitment of RTA to the SRL is facilitated by ribosomal P-stalk proteins, whose C-terminal domains interact with a cavity on RTA normally masked by RTB; however, evidence that this interaction is critical for RTA activity within cells is lacking. Here, we characterized a collection of single-domain antibodies (V
    MeSH term(s) Animals ; Epitopes/metabolism ; Mammals/metabolism ; Peptides/metabolism ; RNA, Ribosomal, 28S/metabolism ; Ribosomal Proteins/chemistry ; Ribosomal Proteins/metabolism ; Ribosomes/metabolism ; Ricin/chemistry ; Single-Domain Antibodies/metabolism
    Chemical Substances Epitopes ; Peptides ; RNA, Ribosomal, 28S ; Ribosomal Proteins ; Single-Domain Antibodies ; Ricin (9009-86-3)
    Language English
    Publishing date 2022-02-17
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1016/j.jbc.2022.101742
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    Uc I Zs.108: Show issues Location:
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    bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
    ab Jg. 2022: Lesesaal (EG)

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  4. Article ; Online: Intracellular Neutralization of Ricin Toxin by Single-domain Antibodies Targeting the Active Site.

    Rudolph, Michael J / Czajka, Timothy F / Davis, Simon A / Thi Nguyen, Chi My / Li, Xiao-Ping / Tumer, Nilgun E / Vance, David J / Mantis, Nicholas J

    Journal of molecular biology

    2020  Volume 432, Issue 4, Page(s) 1109–1125

    Abstract: The extreme potency of the plant toxin, ricin, is due to its enzymatic subunit, RTA, which inactivates mammalian ribosomes with near-perfect efficiency. Here we characterized, at the functional and structural levels, seven alpaca single-domain antibodies ...

    Abstract The extreme potency of the plant toxin, ricin, is due to its enzymatic subunit, RTA, which inactivates mammalian ribosomes with near-perfect efficiency. Here we characterized, at the functional and structural levels, seven alpaca single-domain antibodies (V
    MeSH term(s) Animals ; Antibodies, Neutralizing/immunology ; Antibodies, Neutralizing/metabolism ; Binding Sites, Antibody ; Catalytic Domain ; Chlorocebus aethiops ; Enzyme-Linked Immunosorbent Assay ; Polymerase Chain Reaction ; Ricin/chemistry ; Ricin/immunology ; Single-Domain Antibodies/immunology ; Single-Domain Antibodies/metabolism ; Surface Plasmon Resonance ; Vero Cells
    Chemical Substances Antibodies, Neutralizing ; Single-Domain Antibodies ; Ricin (9009-86-3)
    Language English
    Publishing date 2020-01-10
    Publishing country Netherlands
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 80229-3
    ISSN 1089-8638 ; 0022-2836
    ISSN (online) 1089-8638
    ISSN 0022-2836
    DOI 10.1016/j.jmb.2020.01.006
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    In stock of ZB MED Cologne/Königswinter

    Zs.A 867: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

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  5. Article: Physical and Cognitive Performance of the Least Shrew (Cryptotis parva) on a Calcium-Restricted Diet.

    Czajka, Jessica L / McCay, Timothy S / Garneau, Danielle E

    Behavioral sciences (Basel, Switzerland)

    2012  Volume 2, Issue 3, Page(s) 172–185

    Abstract: ... group of shrews (5 F, 5 M) on a mealworm diet with a calcium concentration comparable to beetle larvae ... collected in the Adirondacks (1.1 ± 0.3 mg/g) and another group (5 F, 3 M) on a mealworm diet with a calcium ...

    Abstract Geological substrates and air pollution affect the availability of calcium to mammals in many habitats, including the Adirondack Mountain Region (Adirondacks) of the United States. Mammalian insectivores, such as shrews, may be particularly restricted in environments with low calcium. We examined the consequences of calcium restriction on the least shrew (Cryptotis parva) in the laboratory. We maintained one group of shrews (5 F, 5 M) on a mealworm diet with a calcium concentration comparable to beetle larvae collected in the Adirondacks (1.1 ± 0.3 mg/g) and another group (5 F, 3 M) on a mealworm diet with a calcium concentration almost 20 times higher (19.5 ± 5.1 mg/g). Animals were given no access to mineral sources of calcium, such as snail shell or bone. We measured running speed and performance in a complex maze over 10 weeks. Shrews on the high-calcium diet made fewer errors in the maze than shrews on the low-calcium diet (F1,14 = 12.8, p < 0.01). Females made fewer errors than males (F1,14 = 10.6, p < 0.01). Running speeds did not markedly vary between diet groups or sexes, though there was a trend toward faster running by shrews on the high calcium diet (p = 0.087). Shrews in calcium-poor habitats with low availability of mineral sources of calcium may have greater difficulty with cognitive tasks such as navigation and recovery of food hoards.
    Language English
    Publishing date 2012-08-16
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2651997-5
    ISSN 2076-328X
    ISSN 2076-328X
    DOI 10.3390/bs2030172
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  6. Article ; Online: Physical and Cognitive Performance of the Least Shrew (Cryptotis parva) on a Calcium-Restricted Diet

    Jessica L. Czajka / Timothy S. McCay / Danielle E. Garneau

    Behavioral Sciences , Vol 2, Iss 3, Pp 172-

    2012  Volume 185

    Abstract: ... group of shrews (5 F, 5 M) on a mealworm diet with a calcium concentration comparable to beetle larvae ... collected in the Adirondacks (1.1 ± 0.3 mg/g) and another group (5 F, 3 M) on a mealworm diet with a calcium ...

    Abstract Geological substrates and air pollution affect the availability of calcium to mammals in many habitats, including the Adirondack Mountain Region (Adirondacks) of the United States. Mammalian insectivores, such as shrews, may be particularly restricted in environments with low calcium. We examined the consequences of calcium restriction on the least shrew (Cryptotis parva) in the laboratory. We maintained one group of shrews (5 F, 5 M) on a mealworm diet with a calcium concentration comparable to beetle larvae collected in the Adirondacks (1.1 ± 0.3 mg/g) and another group (5 F, 3 M) on a mealworm diet with a calcium concentration almost 20 times higher (19.5 ± 5.1 mg/g). Animals were given no access to mineral sources of calcium, such as snail shell or bone. We measured running speed and performance in a complex maze over 10 weeks. Shrews on the high-calcium diet made fewer errors in the maze than shrews on the low-calcium diet (F1,14 = 12.8, p < 0.01). Females made fewer errors than males (F1,14 = 10.6, p < 0.01). Running speeds did not markedly vary between diet groups or sexes, though there was a trend toward faster running by shrews on the high calcium diet (p = 0.087). Shrews in calcium-poor habitats with low availability of mineral sources of calcium may have greater difficulty with cognitive tasks such as navigation and recovery of food hoards.
    Keywords Cryptotis parva ; calcium ; running speed ; spatial memory ; acid deposition ; Neurosciences. Biological psychiatry. Neuropsychiatry ; RC321-571 ; Internal medicine ; RC31-1245 ; Medicine ; R ; DOAJ:Neurology ; DOAJ:Medicine (General) ; DOAJ:Health Sciences ; Psychology ; BF1-990 ; Philosophy. Psychology. Religion ; B ; DOAJ:Psychology ; DOAJ:Social Sciences
    Subject code 590
    Language English
    Publishing date 2012-08-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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