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Lilian C Heck, Matheus V Bianchi, Paula R Pereira, Marina P Lorenzett, Cíntia de Lorenzo, Saulo P Pavarini, David Driemeier, Luciana Sonne
Pythium insidiosum, an aquatic oomycete, causes chronic lesions in the skin and digestive tract of multiple species. A captive-bred Bactrian camel ( Camelus bactrianus) showed clinical signs of lethargy and weight loss in a clinical course of 30 days, with no response to treatment. At necropsy, the abdominal cavity had approximately 32 L of a yellow, turbid fluid with fibrin. The third compartment of the stomach (C-3) showed a focal area of rupture covered with fibrin. Close to this area, the C-3 wall was thickened and firm, demonstrating irregular, yellow, and friable areas on cut surface (kunkers)...
September 2018: Journal of Zoo and Wildlife Medicine: Official Publication of the American Association of Zoo Veterinarians
Uroš Zavrtanik, Junoš Lukan, Remy Loris, Jurij Lah, San Hadži
Truncated versions of heavy chain antibodies (HCAbs) from camelids, also termed nanobodies, comprise only one-tenth the mass of conventional antibodies, yet retain similar, high binding affinities for the antigens. Here we analyze a large dataset of nanobody-antigen crystal structures and investigate how nanobody-antigen recognition compares to the one by conventional antibodies. We find that nanobody paratopes are enriched in aromatic residues just like conventional antibodies, but additionally they also bear a more hydrophobic character...
September 8, 2018: Journal of Molecular Biology
Vladimir Bobkov, Bas van der Woning, Hans de Haard
Variable fragments of heavy-chain-only antibodies (VHH) found in camelids are valuable research tools in pharmacology and biotechnology and are being developed for the clinic to treat patients with autoimmune and infectious diseases or cancer. Their single-domain nature and biochemical properties greatly facilitate the development process. The most common technology to select single-domain antibody fragments is phage display following active immunization of llamas or other members of Camelidae family. Selection of VHH from immune phage libraries is a rapid approach to discover a broad panel of in vivo matured antigen-specific clones with comprehensive functionalities...
2018: Methods in Molecular Biology
Rodrigo A Carrasco, Jaswant Singh, Gregg P Adams
BACKGROUND: A molecule identical to nerve growth factor, with ovulation-inducing properties has been discovered in the seminal plasma of South American camelids (ovulation-inducing factor/nerve growth factor; OIF/NGF). We hypothesize that the ovulatory effect of OIF/NGF is initiated at the level of the hypothalamus, presumably by GnRH neurons. The objective of the present study was to determine the structural relationship between GnRH neurons and neurons expressing high- and low-affinity receptors for NGF (i...
August 31, 2018: Reproductive Biology and Endocrinology: RB&E
Salvador Guardiola, Monica Varese, Macarena Sánchez-Navarro, Cecile Vincke, Meritxell Teixido, Jesús Garcia, Serge Muyldermans, Ernest Giralt
One of the hallmarks of cancer is the overproduction of growth factors such as EGF. Despite the clinical success achieved by EGFR-targeted therapies, their long-term efficacy is compromised by the onset of drug-resistant mutations. To address this issue, we generated a family of camelid-derived single-domain antibodies (Nbs), obtaining the first direct EGF inhibitors that prevent EGFR phosphorylation and pathway activation through this new mechanism of action. The two best Nbs were subjected to a detailed investigation of their interaction mechanism that revealed important differences in their binding kinetics and equilibrium thermodynamics...
August 28, 2018: Angewandte Chemie
Florian Ingo Schmidt
Our understanding of infection biology is based on experiments in which pathogen or host proteins are perturbed by small compound inhibitors, mutation, or depletion. This approach has been remarkably successful, as, for example, demonstrated by the independent identification of the endosomal membrane protein Niemann-Pick C1 as an essential factor for Ebola virus infection in both small compound and insertional mutagenesis screens (Côté, Nature 477:344-348, 2011; Carette et al., Nature 477:340-343, 2011). However, many aspects of host-pathogen interactions are poorly understood because we cannot target all of the involved molecules with small molecules, or because we cannot deplete essential proteins...
2018: Methods in Molecular Biology
Jose A Infantes-Lorenzo, Claire E Whitehead, Inmaculada Moreno, Javier Bezos, Alvaro Roy, Lucas Domínguez, Mercedes Domínguez, Francisco J Salguero
South American camelids are susceptible to tuberculosis, caused mainly by Mycobacterium bovis and M. microti . Despite the tuberculin skin test being the official test for tuberculosis, it has a very low sensitivity in these species (14-20%). Serological tests present the advantages of being rapid, easy to perform and facilitate analysis of large numbers of samples in a short period of time. Novel antigen discovery and evaluation would provide enhanced detection of specific antibodies against members of M. tuberculosis complex...
2018: Frontiers in Veterinary Science
Luise Kultscher, Anja Joachim, Thomas Wittek
OBJECTIVE: The aim of this study was to obtain an overview of management practices, diagnostics, prophylaxis and therapy of endoparasite infections in alpaca herds and of the animal owners perception of this topic, and to combine these data with management data to develop recommendations for prevention and prophylaxis of endoparasitoses in New World camelids. MATERIAL AND METHODS: A questionnaire collected information concerning animal husbandry, feeding and care as well as on prevalence, prophylaxis and therapy of endoparasitoses...
August 2018: Tierärztliche Praxis. Ausgabe G, Grosstiere/Nutztiere
Claudia Abeijon, Julia Dilo, Jacqueline M Tremblay, Agostinho G Viana, Lilian L Bueno, Silvio F G Carvalho, Ricardo T Fujiwara, Charles B Shoemaker, Antonio Campos-Neto
We have recently developed a sensitive and specific urine-based antigen detection ELISA for the diagnosis of visceral leishmaniasis (VL). This assay used rabbit IgG and chicken IgY polyclonal antibodies specific for the Leishmania infantum proteins iron superoxide dismutase 1 (Li-isd1), tryparedoxin1 (Li-txn1) and nuclear transport factor 2 (Li-ntf2). However, polyclonal antibodies have limitations for upscaling and continuous supply. To circumvent these hurdles, we began to develop immortalized monoclonal antibodies...
August 18, 2018: Parasite Immunology
(no author information available yet)
Jessica Li , a fourth year vet student at Cambridge University, spent her EMS placement with Vet Record last week. She used part of her time to delve deeper into the case of Geronimo, the alpaca which tested positive for bovine TB, and the implications for future camelid testing.
August 18, 2018: Veterinary Record
Takeru Suzuki, Yuki Mochizuki, Shinnosuke Kimura, Yoko Akazawa-Ogawa, Yoshihisa Hagihara, Naoto Nemoto
Single-domain antibodies (variable domain of the heavy chain of a heavy chain antibody; VHH) are promising reagents for therapeutics and diagnostics because of their stability, cost-effective production and material workability as a small antibody. Currently, general acquisition of a VHH using immunization of camelids is inconvenient from the standpoint of animal protection, cost and the process is time-consuming. Thus, a straightforward and efficient method for screening VHHs against a target molecule is required...
September 10, 2018: Biochemical and Biophysical Research Communications
Hemant Kumar, Janet S Finer-Moore, Xiaoxu Jiang, Irina Smirnova, Vladimir Kasho, Els Pardon, Jan Steyaert, H Ronald Kaback, Robert M Stroud
The lactose permease of Escherichia coli (LacY), a dynamic polytopic membrane transport protein, catalyzes galactoside/H+ symport and operates by an alternating access mechanism that exhibits multiple conformations, the distribution of which is altered by sugar-binding. Camelid nanobodies were made against a double-mutant Gly46 → Trp/Gly262 → Trp (LacYWW ) that produces an outward-open conformation, as opposed to the cytoplasmic open-state crystal structure of WT LacY. Nanobody 9047 (Nb9047) stabilizes WT LacY in a periplasmic-open conformation...
August 28, 2018: Proceedings of the National Academy of Sciences of the United States of America
Jinxin He, Jiesheng Tian, Junjie Xu, Kai Wang, Ji Li, Shirley J Gee, Bruce D Hammock, Qing X Li, Ting Xu
Variable domain of heavy chain antibody (nanobody, Nb) derived from camelids is an efficient reagent in monitoring environmental contaminants. Oriented conjugates of Nbs and bacterial magnetic particles (BMPs) provide new tools for the high-throughput immunoassay techniques. An anti-tetrabromobisphenol-A (TBBPA) Nb genetically integrated with an extra cysteine residue at the C terminus was immobilized onto BMPs enclosed within the protein membrane, using a heterobifunctional reagent N-succinimidyl-3-(2-pyridyldithiol) propionate, to form a solid BMP-Nb complex...
October 2018: Analytical and Bioanalytical Chemistry
C S Casey, P Orozco-terWengel, K Yaya, M Kadwell, M Fernández, J C Marín, R Rosadio, L Maturrano, D Hoces, Y Hu, J C Wheeler, M W Bruford
Vicuñas and guanacos are two species of wild South American camelids that are key ruminants in the ecosystems where they occur. Although closely related, these species feature differing ecologies and life history characters, which are expected to influence both their genetic diversity and population differentiation at different spatial scales. Here, using mitochondrial and microsatellite genetic markers, we show that vicuña display lower genetic diversity within populations than guanaco but exhibit more structure across their Peruvian range, which may reflect a combination of natural genetic differentiation linked to geographic isolation and recent anthropogenic population declines...
October 2018: Heredity
Laura S Mitchell, Lucy J Colwell
Nanobodies (Nbs) are a class of antigen-binding protein derived from camelid immune systems, which achieve equivalent binding affinities and specificities to classical antibodies (Abs) despite being comprised of only a single variable domain. Here, we use a data set of 156 unique Nb:antigen complex structures to characterize Nb-antigen binding and draw comparison to a set of 156 unique Ab:antigen structures. We analyse residue composition and interactions at the antigen interface, together with structural features of the paratopes of both data sets...
July 25, 2018: Protein Engineering, Design & Selection: PEDS
Aydee Meza, Carina Caldeira, Anthony Valverde, César Ordóñez, Enrique Ampuero, Hernán Cucho, Carles Soler
Semen analysis is a key factor when determining the fertility ability in males. South American camelids, and in particular the alpaca, have been studied very little when compared with other farm animals. The aim of this work was to perform the kinematic characterization of alpaca spermatozoa collected directly from the deferent duct by using CASA-Mot (Computer Assisted Semen Analysis for Motility) technology. Samples were obtained every three days throughout the reproductive season during two periods and with a break of seven days in the middle...
July 25, 2018: Reproduction in Domestic Animals, Zuchthygiene
Kuan-Hui E Chen, Chuan Chen, Tyler Lopez, Kelly C Radecki, Karissa Bustamante, Mary Y Lorenson, Xin Ge, Ameae M Walker
Matrix metalloproteinases (MMPs) are considered excellent targets for cancer therapy because of their important roles in multiple aspects of tumor growth and metastatic spread. However, not all MMPs, or even all activities of specific MMPs, promote cancer. Therefore, there is a need for highly specific inhibitors. Monoclonal antibodies provide the potential for the degree of specificity required, but the isolation of antibodies able to inhibit a specific protease with high selectivity is challenging. Proteolysis specificity lies in recognition of the substrate in or around the active site, which generally forms a concave cleft inaccessible by human IgGs...
June 29, 2018: Oncotarget
Sofia Tabares-da Rosa, Linda A Wogulis, Mark D Wogulis, Gualberto González-Sapienza, David K Wilson
The variable VHH domains of camelid single chain antibodies have been useful in numerous biotechnology applications due to their simplicity, biophysical properties, and abilities to bind to their cognate antigens with high affinities and specificity. Their interactions with proteins have been well-studied, but considerably less work has been done to characterize their ability to bind haptens. A high-resolution structural study of three nanobodies (T4, T9, and T10) which have been shown to bind triclocarban (TCC, 3-(4-chlorophenyl)-1-(3,4-dichlorophenyl)urea) with near-nanomolar affinity shows that binding occurs in a tunnel largely formed by CDR1 rather than a surface or lateral binding mode seen in other nanobody-hapten interactions...
July 23, 2018: Journal of Molecular Recognition: JMR
Lisa C Viesselmann, Ricardo Videla, Bente Flatland
BACKGROUND: The Heska Element POC ("EPOC") is a blood gas instrument intended for use with canine, feline, and equine whole blood; no verification for use with camelid specimens has been reported. OBJECTIVES: Using camelid specimens and commercial quality control materials (QCM), we investigatee EPOC analytical performance and establish EPOC camelid reference intervals (RIs). METHODS: Camelid blood (n = 124) was analyzed using the EPOC (pH, pCO2 , pO2 , HCO3 , base excess, SO2 , sodium, potassium, chloride, ionized calcium, TCO2 , anion gap, HCT, HGB, glucose, lactate, and creatinine); plasma was analyzed using a Roche Cobas c501 (sodium, potassium, chloride, TCO2 , anion gap, glucose, and creatinine)...
September 2018: Veterinary Clinical Pathology
Orleigh A Bogle, Rodrigo A Carrasco, Marcelo H Ratto, Jaswant Singh, Gregg P Adams
The objectives of the study were to compare the presence and localization of OIF/NGF in male reproductive organs and determine the abundance in ejaculates of species representative of both spontaneous- and induced-ovulators. We hypothesized that the protein is a widely conserved component of semen among mammals, but is most abundant in camelids. Immuno-histochemical analysis was performed on tissues from the male reproductive system of llamas, rats, cattle, bison, elk, and white-tailed deer (n = 2 males/species), and the abundance of OIF/NGF in the seminal plasma of camelids (llamas and alpacas), cattle, horses and pigs (n = 69, 53, 24 and 16 ejaculates, respectively) were quantified by radioimmunoassay...
July 2, 2018: Biology of Reproduction
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