Publications

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pseudomonas-associated discospondylitis in a two-month-old llama.a 2-month-old female llama with a history of tetraparesis was presented for necropsy. this cria was apparently normal until it became ataxic during its second week of life. it had diminished flexor reflexes of the left forelimb and both hind limbs. hematology and serum biochemistry revealed neutrophilia, elevated alkaline phosphatase, elevated phosphorus, and modest hyperglycemia. radiography of the cervical spine demonstrated an abnormal c3-c4 intervertebral disc and bony proliferation of the v ...200818460625
seroprevalence of neospora caninum, toxoplasma gondii and sarcocystis sp. in llamas (lama glama) from jujuy, argentina.llamas (lama glama) are south american camelids described as intermediate hosts of neospora caninum, toxoplasma gondii and sarcocystis aucheniae. due to the potential role of these protozoan infections as a cause of economic losses, the aim of this study was to determine the seroprevalence for t. gondii, n. caninum and sarcocystis sp. in llamas from argentina. serum samples from 308 llamas (>2 years old) were collected between 2005 and 2007. a total of 55 farms located in six departments of juju ...200818495344
passive immunization of pigs with bispecific llama single-domain antibody fragments against foot-and-mouth disease and porcine immunoglobulin.foot-and-mouth disease (fmd) is a contagious viral disease of cloven-hoofed animals that occasionally causes outbreaks in europe. we aim to develop an immunotherapy that confers rapid protection against fmd in outbreak situations. for this purpose, we previously isolated llama single-domain antibody fragments (vhhs) binding to fmdv or porcine immunoglobulin (pig). the pig binding vhhs can be genetically fused to other vhhs, resulting in so-called vhh2s. as compared to non-pig binding vhhs such v ...200818534789
alpaca and llama nematodes in britain. 200818567938
polioencephalomalacia in a llama.a 5-year-old, female llama (lama glama) developed acute, progressive neurological disease, characterized by recumbency, muscle fasciculations, intermittent convulsions/opisthotonos, and absent menace responses. postmortem histopathologic lesions, limited to the cerebral cortex, consisted of necrosis of the superficial and deep laminae. the clinical disease and microscopic lesions were consistent with polioencephalomalacia.200818624072
egf induces coalescence of different lipid rafts.the suggestion that microdomains may function as signaling platforms arose from the presence of growth factor receptors, such as the egfr, in biochemically isolated lipid raft fractions. to investigate the role of egfr activation in the organization of lipid rafts we have performed flim analyses using putative lipid raft markers such as ganglioside gm1 and glycosylphosphatidylinositol (gpi)-anchored gfp (gpi-gfp). the egfr was labeled using single domain antibodies from llama glama that specific ...200818628305
cloning, expression, and characterization of a single-domain antibody fragment with affinity for 15-acetyl-deoxynivalenol.a single-domain variable heavy chain (v(h)h) antibody fragment specific to the mycotoxin 15-acetyldeoxynivalenol (15-acdon) was obtained after immunization of a llama (llama glama) with the protein conjugate 15-don-bsa plus titermax classic adjuvant. after confirmation of a polyclonal response to don toxin in both conventional (cigg) and heavy chain antibody (hcab) fractions, a v(h)h library was constructed from amplified cdna by nested pcr. v(h)h fragments with binding affinity for the mycotoxi ...200818632156
llama-derived single-chain antibody fragments directed to rotavirus vp6 protein possess broad neutralizing activity in vitro and confer protection against diarrhea in mice.group a rotavirus is one of the most common causes of severe diarrhea in human infants and newborn animals. rotavirus virions are triple-layered particles. the outer capsid proteins vp4 and vp7 are highly variable and represent the major neutralizing antigens. the inner capsid protein vp6 is conserved among group a rotaviruses, is highly immunogenic, and is the target antigen of most immunodiagnosis tests. llama-derived single-chain antibody fragments (vhh) are the smallest molecules with antige ...200818632867
alpaca and llama nematodes in britain. 200818660528
anaerobic co-digestion of aquatic flora and quinoa with manures from bolivian altiplano.quinoa stalk (chenopodium quinoa willd.) from agricultural crop residue, totora (schoenoplectus tatora) and o-macrophytes (aquatic flora) from lake titicaca (on the bolivian altiplano) were studied in a wet anaerobic co-digestion process together with manure from llama, cow and sheep. anaerobic semi-continuous experiments were performed in (10) 2-l reactors at a temperature of 25 degrees c with 30 days of hydraulic retention time (hrt) and an organic loading rate (olr) of 1.8 kg vs m(-3) d(-1). ...200818155895
development of elisas for the measurement of igm and igg subclasses in sera from llamas (lama glama) and assessment of the humoral immune response against different antigens.members of the camelidae family possess a functional class of antibodies devoid of light chains (known as heavy chain antibodies, hcabs). three igg isotypes have been identified (igg(1), igg(2) and igg(3)); igg(2) and igg(3) are hcabs whereas the igg(1) has the conventional structure. different subtypes of igg(1) (igg(1a) and igg(1b)) and igg(2) (igg(2a), igg(2b) and igg(2c)) have been classified according to variations in the amino acids sequence of the hinge region. the single variable domain ...200818692907
the synergizer service for translating gene, protein and other biological identifiers.the synergizer is a database and web service that provides translations of biological database identifiers. it is accessible both programmatically and interactively. availability: the synergizer is freely available to all users inter-actively via a web application (http://llama.med.harvard.edu/synergizer/translate) and programmatically via a web service. clients implementing the synergizer application programming interface (api) are also freely available. please visit http://llama.med.harvard.ed ...200818697767
tnt detection using llama antibodies and a two-step competitive fluid array immunoassay.llamas possess unique subclasses of antibodies that lack light chains, and thus are made by the pairing of two heavy chains. igg was purified from two llamas which had been immunized with trinitrobenzene-keyhole limpet hemocyanin. conventional igg1 and heavy chain igg2 and igg3 subclasses were fractionated using affinity chromatography. the effectiveness of heavy chain antibodies for the detection of trinitrotoluene (tnt) using a competitive fluid array immunoassay was evaluated and compared to ...200818755196
conditional inactivation of hif-1 using intrabodies.hypoxia is a hallmark of solid cancers and triggers the transcription of genes responsible for cell survival. the transcription factor hypoxia-inducible factor 1 (hif-1) is a key regulator in this response and frequently activated in human cancer. hif-1 activation is associated with tumor aggressiveness and poor clinical outcome and, therefore, may provide an attractive therapeutic target. here we provide a novel approach for hif-1 targeted therapy using single-domain llama antibodies directed a ...200818791271
design of a village breeding programme for a llama population in the high andes of bolivia.no national breeding programme for llamas is in place in bolivia. initiatives for genetic improvement are rarely found and are usually carried out by ngos working in rural development or improvement of livestock production or research stations. farmers in the province of ayopaya in the district of cochabamba have formed a breeders' organization with the aim of improving fibre production. in this study, a detailed outline of a breeding programme with a focus on organizational and technical detail ...200818803786
llama antibody fragments with cross-subtype human immunodeficiency virus type 1 (hiv-1)-neutralizing properties and high affinity for hiv-1 gp120.members of the camelidae family produce immunoglobulins devoid of light chains. we have characterized variable domains of these heavy chain antibodies, the vhh, from llamas immunized with human immunodeficiency virus type 1 (hiv-1) envelope protein gp120 in order to identify vhh that can inhibit hiv-1 infection. to increase the chances of isolating neutralizing vhh, we employed a functional selection approach, involving panning of phage libraries expressing the vhh repertoire on recombinant gp12 ...200818842738
simple sequence repeats for genetic studies of alpaca.trace sequences from the 2x alpaca genome sequencing effort were examined to identify simple sequence repeats (microsatellites) for genetic studies. a total of 6,685 repeat-containing sequences were downloaded from genbank, processed, and assembled into contigs representing an estimated 4,278 distinct sequences. this sequence set contained 2,290 sequences of length > 100 nucleotides that contained microsatellites of length > or = 14 dinucleotide or 10 trinucleotide repeats with purity equal to 1 ...200818855250
thermostable llama single domain antibodies for detection of botulinum a neurotoxin complex.immunoglobulins from animals of the camelidae family boast unique forms that do not incorporate light chains. antigen binding in these unconventional heavy-chain homodimers is mediated through a single variable domain. when expressed recombinantly these variable domains are termed single domain antibodies (sdab) and are among the smallest naturally igg-derived antigen binding units. sdab possess good solubility, thermostability, and can refold after heat and chemical denaturation making them pro ...200818947189
relevance of the diversity among members of the trypanosoma cruzi trans-sialidase family analyzed with camelids single-domain antibodies.the sialic acid present in the protective surface mucin coat of trypanosoma cruzi is added by a membrane anchored trans-sialidase (tcts), a modified sialidase that is expressed from a large gene family. in this work, we analyzed single domain camelid antibodies produced against trans-sialidase. llamas were immunized with a recombinant trans-sialidase and inhibitory single-domain antibody fragments were obtained by phage display selection, taking advantage of a screening strategy using an inhibit ...200818949046
placental glycosylation in a cama (camel-llama cross) and its relevance to successful hybridisation. 200818950721
parasite-induced leucomyelopathy in llamas (lama glama). 200818953077
probing of actinobacillus pleuropneumoniae apxiiia toxin-dependent cytotoxicity towards mammalian peripheral blood mononucleated cells.abstract:200819046441
infertility associated with persistent hymen in an alpaca and a llama.perforation and dilation of the persistent hymen in an alpaca and a llama, detected by vaginal examination and endoscopy, was achieved by use of a sigmoidoscope and incremental dilation using cylindrical instruments to a maximum diameter of 38 mm. outcome and subsequent fertility are dependent on length of time the obstruction has been present and secondary uterine disease.200819183735
immunological applications of single-domain llama recombinant antibodies isolated from a naïve library.we describe the rapid isolation of single-domain recombinant antibodies in vhh format from a pre-immune llama library created in our laboratory. such naïve library has demonstrated to be a versatile tool and enabled the isolation of several different antibodies for any of the six proteins panned in parallel. the binders specific for human fibroblast growth factor receptor 1 (fgfr1) were successively analyzed in more detail and resulted suitable for both western blot and immunofluorescence analys ...200919196718
platelet aggregation responses in clinically healthy adult llamas.limited information exists regarding hemostasis in camelids despite the importance of platelet function testing in the accurate identification of platelet disorders. as further importation of llamas to north america is restricted, variability in breeding stock will continue to decrease, potentially leading to an increase in heritable bleeding disorders.200919228355
prevention of oculopharyngeal muscular dystrophy by muscular expression of llama single-chain intrabodies in vivo.oculopharyngeal muscular dystrophy (opmd) is a late onset disorder characterized by progressive weakening of specific muscles. it is caused by short expansions of the n-terminal polyalanine tract in the poly(a) binding protein nuclear 1 (pabpn1), and it belongs to the group of protein aggregation diseases, such as huntington's, parkinson's and alzheimer diseases. mutant pabpn1 forms nuclear aggregates in diseased muscles in both patients and animal models. intrabodies are antibodies that are mod ...200919258344
pentabody-mediated antigen delivery induces antigen-specific mucosal immune response.an efficient immunization system is essential for the development of mucosal vaccine. cholera toxin (ct) and escherichia coli heat labile toxin (lt) are among the strongest adjuvants tested in experimental animals but their use in humans has been hindered by their toxicity. on the other hand, the role of their non-toxic b-subunits, ctb or ltb, in enhancing mucosal immune response is not clear. we propose here a novel strategy for the induction of mucosal immune responses. single domain antibodie ...200919269688
cotoneaster sp. poisoning in a llama (lama glama).a case of poisoning with a cotoneaster sp. is described in a llama (lama glama). the clinical signs were dyspnea, with congested mucous membranes, chewing, recumbency, and opisthotonos. several hours after ingestion of the plant, the llama died, despite treatment with sodium thiosulfate and nitrite. necropsy revealed approximately 676 g of leaves and fruit of cotoneaster in the first compartment of the stomach. the blood was light red and did not clot. because cotoneaster sp. contains low concen ...200919286507
metastatic uterine adenocarcinoma and hepatic lipomatosis in a llama (lama glama).a 6-year-old, female llama (lama glama) presented with progressive anorexia and ascites. postmortem examination revealed an infiltrative uterine adenocarcinoma with widespread metastases. the neoplasm completely replaced and infiltrated the myometrium of the uterine body and cervix and metastasized largely to the serosal surfaces of the peritoneal cavity. histopathology identified a highly invasive growth of neoplastic cells in solid packets or tubular arrangements and marked fibroplasia. no bov ...200919286516
prevalence of eimeria macusaniensis and eimeria ivitaensis in south american camelids of northwest argentina.faecal samples from mostly adult llamas (n=626), vicuñas (n=161) and guanacos (n=4) were obtained between december 2004 and july 2008 in three provinces of northwest argentina in order to study the prevalence of eimeria macusaniensis and eimeria ivitaensis. faeces were examined by a flotation technique using a cl(2)zn+clna solution (specific gravity=1.59). oocysts of e. macusaniensis occurred in 88.3% of 77 llama herds and in 50.3% of 626 llamas sampled whereas oocysts of e. ivitaensis occurred ...200919349120
crystallization of the altitude adapted hemoglobin of guinea pig.hemoglobin is the versatile oxygen carrier in the blood of vertebrates and a key factor for adaptation to live in high altitudes. several structural changes are known to account for increased oxygen affinity in hemoglobin of altitude adapted animals such as llama and barheaded goose. guinea pigs are adapted to live in high altitudes in the andes and consequently their hemoglobin has an increased oxygen affinity. however, the structural changes responsible for the adaptation of guinea pig hemoglo ...200919356144
reverse proteomic antibody screening identifies anti adhesive vhh targeting vla-3.therapeutic approaches aimed at targeting tumor surface markers using monoclonal antibodies provide a powerful strategy in cancer treatment. here we report selection of single variable domains (vhh) of llama heavy chain antibodies, using a vhh-phage-display library. a reverse proteomic approach was used to identify the cognate proteins recognized by enriched vhh on hela cells. one of these vhh bound the integrin alpha 3 beta 1 (vla-3) and was further characterized. most interestingly, this vhh c ...200919359042
passive immunization with llama single-domain antibody fragments reduces foot-and-mouth disease transmission between pigs.we aim to develop a method that confers rapid protection against foot-and-mouth disease (fmd) by passive immunization with recombinant llama single-domain antibody fragments (vhhs). previously constructed genetic fusions of two vhhs (vhh2s) that either neutralizes fmdv or binds to porcine immunoglobulin to increase the serum half-life, conferred only limited protection to pigs. we therefore now generated vhh3s containing an additional fmdv binding vhh. two vhh3s neutralized fmdv more potently th ...200919368770
effect of ecg superstimulation and buserelin on cumulus-oocyte complexes recovery and maturation in llamas (lama glama).the aim of the present study was two-fold. experiment i: evaluate the effect of buserelin on llama's oocyte maturation after exogenous follicular activity suppression, followed by ovarian superstimulation with different doses of equine chorionic gonadotropin (ecg). experiment ii: compare the number of follicles aspirated and the number of cumulus-oocyte complexes (cocs) recovered according to different doses of ecg followed by buserelin. experiment i consisted in a control group (without buserel ...200919392664
mange in alpacas, llamas and goats in the uk: incidence and risk.a retrospective postal questionnaire was used to obtain information about the prevalence of mange and its association with husbandry-related risk factors, in alpaca, llama and goat herds in the uk. in total 1797 questionnaires were sent out to members of the british alpaca society, the british llama society and the british goat society, giving response rates of 40.4%, 29.3% and 22.8% from the three groups, respectively. between january and december 2007, mange was reported in 52.2% (151 of 292), ...200919446959
alpaca and llama health management. preface. 200919460637
management of neonatal llamas and alpacas.most llama and alpaca crias will be born without complication and survive the neonatal period without incident. however, it is important to be able to recognize which crias are likely to be at risk of complications so that you are best able to advise owners and take the correct course of action if required. this article deals with management of the pregnant camelid, the events associated with parturition and the peripartum period with emphasis on the cria, management of the newborn cria includin ...200919460645
chemical restraint, anesthesia, and analgesia for camelids.this article covers techniques used to provide chemical restraint, injectable or inhalation anesthesia, and analgesia in camelid patients. understanding the information presented improves safety and efficacy when using the techniques presented in the article. this article focuses primarily on llama and alpaca patients. of the techniques that have been used on camels, the dosing protocols provided have proven effective.200919460650
specific immuno capturing of the staphylococcal superantigen toxic-shock syndrome toxin-1 in plasma.toxic-shock syndrome is primarily caused by the toxic-shock syndrome toxin 1 (tsst-1), which is secreted by the gram-positive bacterium staphylococcus aureus. the toxin belongs to a family of superantigens (sags) which exhibit several shared biological properties, including the induction of massive cytokine release and v(beta)-specific t-cell proliferation. in this study we explored the possibility to use monoclonal variable domains of llama heavy-chain antibodies (vhh) in the immuno capturing o ...200919475676
a llama single domain anti-idiotypic antibody mimicking her2 as a vaccine: immunogenicity and efficacy.her2 over-expression in breast cancer correlates with reduced survival. anti-idiotypic antibodies (abs) that closely mimic her2 could play a crucial role in the development of effective therapeutic vaccines. here, we show that an anti-idiotypic single domain antibody (sdab) 1he isolated from a library generated from a trastuzumab f(ab')(2)-immunized llama, closely mimics her2. sdab 1he shows high affinity for trastuzumab f(ab')(2), selectively inhibits her2 binding to trastuzumab f(ab')(2), and ...200919523913
llama single-domain antibodies directed against nonconventional epitopes of tumor-associated carcinoembryonic antigen absent from nonspecific cross-reacting antigen.single-domain antibodies (sdabs), which occur naturally in camelids, are endowed with many characteristics that make them attractive candidates as building blocks to create new antibody-related therapeutic molecules. in this study, we isolated from an immunized llama several high-affinity sdabs directed against human carcinoembryonic antigen (cea), a heavily glycosylated tumor-associated molecule expressed in a variety of cancers. these llama sdabs bind a different epitope from those defined by ...200919531051
capillary electrophoresis of intact basic proteins using noncovalently triple-layer coated capillaries.the usefulness of a noncovalent, positively charged capillary coating for the efficient analysis of intact basic proteins with ce was studied. capillaries were coated by subsequent flushing with solutions of 10% w/v polybrene (pb), 3% w/v dextran sulfate (ds), and again 10% w/v pb. coating characterization studies showed that stable coatings could be produced which exhibited a ph-independent and highly reproducible eof. the pb-ds-pb coating was evaluated with tris phosphate bges of various ph us ...200919557816
mycobacterium microti--pulmonary tuberculosis in an immunocompetent patient.human tuberculosis is caused by members of the mycobacterium tuberculosis complex, which includes m. tuberculosis, m. bovis, m. africanum and m. bovis bcg. however there are increasing reports of rarely occurring genetic variants such as m. canettii, m. microti and m. pinipedii. the natural reservoir, mode of transmission and potential modification of host interaction of these species is not yet fully elucidated. we report a rare case of extensive cavitary smear-positive tuberculosis of the left ...200919562286
llama vhh antibody fragments against gfap: better diffusion in fixed tissues than classical monoclonal antibodies.camelids produce antibodies made of homodimeric heavy chains, and the antigen-binding region being composed of a single domain called vhh. these vhhs are much smaller than complete igg. they are also more thermostable and more soluble in water; they should, therefore, diffuse more readily in the tissues. vhhs, expressed in bacteria, are easier to produce than conventional monoclonal antibodies. because of these special characteristics, these antibody fragments could have interesting developments ...200919597828
ovarian response and embryo production in llamas treated with equine chorionic gonadotropin alone or with a progestin-releasing vaginal sponge at the time of follicular wave emergence.the objective of the study was to compare the ovulatory response and embryo production in llamas (lama glama) treated with a single dose of equine chorionic gonadotropin (ecg) alone or combined with intravaginal medroxyprogesterone acetate (mpa) at the time of follicular wave emergence. llamas with a growing follicle >or=7 mm in diameter were assigned to one of the following groups: (1) control (n=28): nonstimulated llamas were mated and embryos were collected 7 d after mating. (2) ecg (n=32): l ...200919616837
oleander intoxication in new world camelids: 12 cases (1995-2006).to characterize the clinical and clinicopathologic effects and evaluate outcome associated with oleander toxicosis in new world camelids.200919650704
protein mislocalization in plant cells using a gfp-binding chromobody.a key challenge in cell biology is to directly link protein localization to function. the green fluorescent protein (gfp)-binding protein, gbp, is a 13-kda soluble protein derived from a llama heavy chain antibody that binds with high affinity to gfp as well as to some gfp variants such as yellow fluorescent protein (yfp). a gbp fusion to the red fluorescent protein (rfp), a molecule termed a chromobody, was previously used to trace in vivo the localization of various animal antigens. in this st ...200919686537
next generation software for functional trend analysis.funcassociate is a web application that discovers properties enriched in lists of genes or proteins that emerge from large-scale experimentation. here we describe an updated application with a new interface and several new features. for example, enrichment analysis can now be performed within multiple gene- and protein-naming systems. this feature avoids potentially serious translation artifacts to which other enrichment analysis strategies are subject. availability: the funcassociate web applic ...200919717575
a single-step procedure of recombinant library construction for the selection of efficiently produced llama vh binders directed against cancer markers.heavy chain antibodies are naturally occurring in camelidae (camels and llamas). their variable domain (vhh) can be efficiently produced as a recombinant protein in e. coli with a large range of applications in the fields of diagnostics and immunotherapy. standard cloning approach involves resolution of vhh from the heavy chain variable domain of conventional antibodies (vh) by a nested pcr amplification followed by a phage display based selection. present work illustrates that in contrast to us ...200919744487
use of serological techniques for diagnosis of mycobacterium bovis infection in a llama herd. 200919749210
antibody response to the epsilon toxin of clostridium perfringens following vaccination of lama glama crias.enterotoxaemia produced by clostridium perfringens a, c and d is an important cause of mortality in young llamas. there is no data on antibody responses following vaccination with epsilon toxin.200919801806
lamanema chavezi (nematoda: molineidae): epidemiological data of the infection in south american camelids of northwest argentina.faecal samples from llamas (n=708), vicuñas (n=171) and guanacos (n=4) were obtained between december 2004 and may 2009 in three provinces of northwest argentina (jujuy, salta and catamarca) to know the distribution, prevalence and intensity of lamanema chavezi infection in these south american camelid species (sacs). faeces were examined by a sedimentation-flotation technique using a cl(2)zn+clna solution (specific gravity=1.59). eggs of l. chavezi occurred in 30.3% of 89 llama herds and in 18. ...200919819627
isolation of monoclonal antibody fragments from phage display libraries.techniques developed over the past 20 years for the display of foreign peptides and proteins on the surfaces of filamentous bacteriophages have been a major driving force in the rapid development of recombinant antibody technology in recent years. with phage display of antibodies as one of its key components, recombinant antibody technology has led to the development of an increasing number of therapeutic monoclonal antibodies. antibody gene libraries are fused to a gene encoding a phage coat pr ...200919082566
llama-derived single-domain intrabodies inhibit secretion of hepatitis b virions in mice.hepatitis b virus (hbv) infections cause 500,000 to 700,000 deaths per year as a consequence of chronic hepatitis, cirrhosis, and hepatocellular carcinoma. efficient and safe antivirals to treat chronically infected patients and consequently to prevent development of hepatocellular carcinoma are still awaited. we isolated five single-domain antibodies (vhhs) that recognize the most abundant envelope protein (s) of hbv. vhhs, when expressed and retained in the endoplasmic reticulum as intrabodies ...200919085971
reproductive glycogenetics--a critical factor in pregnancy success and species hybridisation.hybridisation occurs rarely in nature and experiments using interspecific transfer of embryos generally result in implantation failure. here we show that appropriate glycosylation of the apposing surfaces of endometrium and trophoblast probably is an important factor and may play a critical role in pregnancy success. examination of closely related species shows that each has its own specific pattern of glycosylation, or glycotype, at the fetomaternal interface and that interacting surfaces appea ...200919121542
simulated mutagenesis of the hypervariable loops of a llama vhh domain for the recovery of canonical conformations.in this work, wildtype and mutated hypervariable regions of an anti-hcg llama vhh antibody were simulated via a molecular dynamics replica exchange method (rem). seven mutants were simulated with the goal of identifying structural determinants that return the noncanonical h1 loop of the wildtype antibody to the type 1 canonical structure predicted by database methods formulated for conventional antibodies. two cases with three point mutations yielded a stable type 1 h1 structure. in addition, ot ...200919132876
isolation of novel egfr-specific vhh domains.epidermal growth factor receptor (egfr) is overexpressed or mutated in a high percentage of tumors. egfr has long been considered a promising target for cancer diagnostic and therapeutic applications. however, monoclonal antibodies and other large antibody constructs diffuse into tumors slowly, limiting their efficacy. to develop lower molecular weight probes for egfr and other tumor cell receptors, the authors immunized a llama with the extracellular domains (ecds) of egfr and an oncogenic muta ...200919171923
pre-weaning morbidity and mortality of llamas and alpacas.to describe the morbidity and mortality patterns and identify factors associated with morbidity in pre-weaning llamas and alpacas.200919178480
conformation and anatomical relations of the liver of llama (lama glama).morphological studies of the liver of the llama are structural supportive to the clinical practice, surgery and specific diagnostic techniques. the aims of this study were first to determine the location of the organ and the direction of its major axis to project it to the abdominal wall, identifying visible and palpable bony references. secondly, to characterize and determine anatomical relations of the surfaces, borders and angles of the llama liver, as well as, of its lobulation. twenty adult ...200919183352
glycosylation at the fetomaternal interface: does the glycocode play a critical role in implantation?during pregnancy, the heavily glycosylated surfaces of the implanting blastocyst and maternal uterine epithelium interact in a highly controlled and specific manner. examination of this interface in species that show interdigitation of embryonic and maternal surfaces (epitheliochorial placentation) shows that each has its own particular pattern of glycosylation or glycotype, and that closely related and/or interbreeding species e.g. horse and donkey or llama and guanaco, have very similar glycot ...200918677581
a one-step exclusion-binding procedure for the purification of functional heavy-chain and mammalian-type gamma-globulins from camelid sera.a new approach has recently been proposed for the purification of 'mammalian-type' igg, consisting of exclusion binding. the technique uses a gel ('melon gel'; pierce) that binds to all plasma proteins, but not to iggs, thus allowing iggs to be recovered in the ft (flow-through) fraction. here, the technique was applied to camelid iggs, which are known to be composed of not only classic mammalian-type iggs (igg1) but also hc-iggs (heavy chain iggs). both mammalian type and hc-iggs can be purifie ...200919824883
[identification of immunoreactive antigens of leptospira interrogans].a batch of 28 llama (lama gama) sera from jujuy province in argentina was studied in order to identify immune reactive antigens to leptospira interrogans. different antigenic preparations from the bacterium were used to study the immune reactivity by the microagglutination (mat), elisa and western immunoblot tests. a control pool of positive bovine sera was used. all the llama sera were negative to mat as well as to elisa. two of the llama sera and the positive bovine sera pool rendered positive ...200919831309
differential recognition of vascular and parenchymal beta amyloid deposition.by phage display, llama-derived heavy chain antibody fragments were selected from non-immune and immune libraries and tested for their affinity and specificity for beta amyloid by phage-elisa, immunohistochemistry and surface plasmon resonance. we identified eight distinct heavy chain antibody fragments specific for beta amyloid. while three of them recognized vascular and parenchymal beta amyloid deposits, the remaining five heavy chain antibody fragments recognized vascular beta amyloid specif ...200920015576
in vivo and in vitro sperm interaction with oviductal epithelial cells of llama.sperm reservoirs in south american camelids would be crucial for successful fertilization. since ovulation occurs approximately 36 h after mating, the maintenance of the sperm viability in the oviduct waiting for the ovum is a critical reproductive event. our study aimed at determining whether the isthmus or the utero tubal junction (utj) could function as a sperm reservoir in llama by means of in vivo and in vitro experiments. for the in vivo experiments, the oviducts of adult females with a do ...200920468081
immunohistochemical study of rabies virus within the central nervous system of domestic and wildlife species.immunohistochemistry using a commercial polyclonal antibody for lyssavirus was applied to 39 archival cases of rabies. paraffin blocks from 13 different species were available, including 3 dogs, 4 cats, 1 pig, 6 cattle, 4 horses, 1 llama, 7 skunks (mephitis mephitis), 7 raccoons (procyon lotor), 1 bat (myotis species), 1 white-tailed deer (odocoileus virginianus), 1 bobcat (lynx rufus), 2 gray foxes (urocyon cinereoargenteus), and 1 red fox (vulpes vulpes). all cases had previously been diagnose ...201020484176
the complete mitochondrial dna sequence of the guanaco (lama guanicoe): comparative analysis with the vicuña (vicugna vicugna) genome.south american camelids comprise the guanaco (lama guanicoe) and the vicuña (vicugna vicugna), which are wild species, and the domestic llama (lama glama) and alpaca (lama pacos). this paper presents the first complete mitochondrial (mt) genome of the guanaco and the mt coding sequence of the vicuña. the guanaco mtdna is 16,649 nt long and its composition and organization are similar to the mitochondrial genome of other mammals. excluding the control region, comparison of the complete guanaco an ...201020524143
confirmation of the chewing louse, bovicola breviceps, in a british llama (lama glama) herd. 201020562379
llama-derived single-domain antibodies for the detection of botulinum a neurotoxin.single-domain antibodies (sdab) specific for botulinum neurotoxin serotype a (bont a) were selected from an immune llama phage display library derived from a llama that was immunized with bont a toxoid. the constructed phage library was panned using two methods: panning on plates coated with bont a toxoid (bont a td) and bont a complex toxoid (bont ac td) and panning on microspheres coupled to bont a td and bont a toxin (bont a tx). both panning methods selected for binders that had identical se ...201020582697
kinetic analysis of novel mono- and multivalent vhh-fragments and their application for molecular imaging of brain tumours.the overexpression of epidermal growth factor receptor (egfr) and its mutated variant egfrviii occurs in 50% of glioblastoma multiforme. we developed antibody fragments against egfr/egfrviii for molecular imaging and/or therapeutic targeting applications.201020590596
embryo transfer technique: factors affecting the viability of the corpus luteum in llamas.the aim of the present study was to evaluate the effect of the embryo transfer (et) maneuvers on plasma progesterone concentrations in recipient lama glama females and the relationship between the site the embryo was transferred to and corpus luteum (cl) localization. experiment i (effect of transcervical threading): adult non-pregnant, non-lactating llama females were randomly assigned into two groups: control group (without cervical threading, n=10) and group a (with cervical threading, n=10). ...201020663621
esophageal dysfunction in four alpaca crias and a llama cria with vascular ring anomalies.3 alpaca crias and cadavers of an alpaca cria and a llama cria were evaluated for evidence of esophageal dysfunction.201020673113
antibodies to pathogenic livestock viruses in a wild vicuña (vicugna vicugna) population in the argentinean andean altiplano.serum samples from 128 wild vicuñas (vicugna vicugna) were tested for antibodies (ab) to rotavirus (rv), bovine parainfluenza virus 3 (bpiv-3), bovine herpesvirus-1 (bhv-1), bovine viral diarrhea virus (bvdv-1), foot-and-mouth disease virus (fmdv), bluetongue virus (btv), equine herpesvirus-1 (ehv-1), and influenza a virus equine (eiv). samples were collected in cieneguillas province of jujuy, in northern argentina. feces from 44 vicuñas were also collected to investigate rv shedding. llamas (la ...201020688660
skin lesions associated with demodex sp. in a llama (lama glama). [corrected]a 2.5-yr-old female llama (lama glama) [corrected] with skin lesions was presented to the animal health center in seoul grand park zoo, korea. mites of the genus demodex in the absence of other mites or fungi were identified from the lesions by skin scrapings. the bodies were elongated, tapered, and 200-280 1m in length; four pairs of stumpy legs were present at the front of the body, and the striated opisthosoma constituted about half of the body length. histologic examination of the skin biops ...201020722277
characterization of vancomycin-resistant enterococcus faecium isolated from swine in three michigan counties.vancomycin-resistant enterococci are a major cause of nosocomial infections but are rarely found in humans in the community and have not been identified in food animals in the united states. we evaluated a total of 360 fecal specimens from humans and their animals being raised for exhibit at three county fairs in michigan. fecal samples from 158 humans, 55 swine, 50 cattle, 25 horses, 57 sheep, 14 goats, and 1 llama were obtained and plated onto enterococcosel agar containing 16 μg/ml of vancomy ...201020739498
prevalence of mycoplasma haemolamae infection in peruvian and chilean llamas and alpacas.mycoplasma haemolamae is a hemotropic mycoplasma that affects red blood cells of llamas (lama glama) and alpacas (lama pacos). it is variably associated with anemia, and most infections are subclinical. development of a polymerase chain reaction assay has facilitated detection of this infection in llamas and alpacas in the united states and other countries. whether the infection occurs in camelids in south america has previously been unknown. the current study documents a 15.8% infection rate am ...201020807939
mast cell tumors in a llama (lama glama).a 9-year-old female llama (lama glama) that served as a blood donor at the ohio state university veterinary teaching hospital developed multiple small, raised, firm, non-haired cutaneous masses on the right hip, left cheek, and right and left shoulders. cytological evaluation of fine-needle aspirates from the cutaneous mass from the left shoulder and right hip comprised many well-differentiated, highly granulated mast cells with moderate numbers of eosinophils. occasional mast cells exhibited er ...201020807950
tandem affinity depletion: a combination of affinity fractionation and immunoaffinity depletion allows the detection of low-abundance components in the complex proteomes of body fluids.protein biomarker discovery in the low concentration range of human body fluids requires the enrichment of the proteins of interest. here we report on a tandem affinity strategy: in the first step, we isolated a human plasma glyco-subproteome of healthy individuals by wheat germ agglutinin (wga) lectin affinity chromatography. in the second step, the proteins of this subproteome were used to raise antibodies in llama (lama glama). the heavy-chain fraction of the llama antibodies was used to depl ...201020839810
assessment of antemortem tests used in the control of an outbreak of tuberculosis in llamas (lama glama).an outbreak of tuberculosis (tb) caused by mycobacterium bovis in a llama herd is described. over a 25-month period, a total of 70 llamas were selected for postmortem examination using four distinct criteria: clinical suspicion of disease (15 animals), positive tuberculin skin test result (three animals), antibody positive using a novel serological test (rapid test, 54 animals) and elective cull (five animals). some animals qualified on more than one criterion. gross lesions of tb were detected ...201020871080
ovarian function in south american camelids (alpacas, llamas, vicunas, guanacos).ultrasound technology and hormone assays have provided a better understanding of folliculogenesis and ovulation in south american camelids in the last two decades. females exhibit waves of ovarian follicular growth and are induced ovulators and therefore do not exhibit oestrous cycles in the manner of spontaneously ovulating species such as sheep and cattle. there is much variation in inter-wave interval among camelid species (alpaca/llama 10-22 days, vicuna 4-11 days), within species and within ...201020875933
single-domain llama antibodies as specific intracellular inhibitors of spvb, the actin adp-ribosylating toxin of salmonella typhimurium.adp-ribosylation of host cell proteins is a common mode of cell intoxication by pathogenic bacterial toxins. antibodies induced by immunization with inactivated adp-ribosylating toxins provide efficient protection in case of some secreted toxins, e.g., diphtheria and pertussis toxins. however, other adp-ribosylating toxins, such as salmonella spvb toxin, are secreted directly from the salmonella-containing vacuole into the cytosol of target cells via the spi-2 encoded bacterial type iii secretio ...201020940265
molecular imaging of glioblastoma multiforme using anti-insulin-like growth factor-binding protein-7 single-domain antibodies.insulin-like growth factor-binding protein 7 (igfbp7) is an abundant, selective and accessible biomarker of glioblastoma multiforme (gbm) tumour vessels. in this study, an anti-igfbp7 single-domain antibody (sdab) was developed to target gbm vessels for molecular imaging applications.201020959824
structures of a key interaction protein from the trypanosoma brucei editosome in complex with single domain antibodies.several major global diseases are caused by single-cell parasites called trypanosomatids. these organisms exhibit many unusual features including a unique and essential u-insertion/deletion rna editing process in their single mitochondrion. many key rna editing steps occur in ∼20s editosomes, which have a core of 12 proteins. among these, the "interaction protein" krepa6 performs a central role in maintaining the integrity of the editosome core and also binds to ssrna. the use of llama single do ...201020969962
chronic myositis in an australian alpaca (llama pacos) associated with sarcocystis spp.an alpaca (llama pacos), born and raised in australia, was presented with multiple subcutaneous abscesses. histological findings indicated a severe necrotizing and histiocytic myositis and cellulitis associated with central caseation and multiple sarcocysts. ultrastructural examination supported the diagnosis; however, cyst wall characteristics were not consistent with the 2 known species found in alpacas. while seroconversion in camelids is reported to be near ubiquitous, myositis is rare, and ...201021088185
evaluation of a postexposure rabies prophylaxis protocol for domestic animals in texas: 2000-2009.to determine whether postexposure rabies prophylaxis (pep) in domestic animals, as mandated in texas, has continued to be effective and to evaluate preexposure or postexposure vaccination failures from 2000 through 2009.201021155679
comparison of fixation methods for treatment of long bone fractures in llamas and alpacas.to determine outcome after repair of long bone fractures in llamas and alpacas.201021175689
granulosa theca cell tumor with erythrocytosis in a llama.a 2.5-year-old, female llama with weight loss and lethargy had a packed cell volume (pcv) of 45% which increased to 57% over 3 wk. transrectal ultrasonography revealed a mass of mixed echogenicity involving the right ovary, which was removed. a histopathological diagnosis of granulosa theca cell tumor was made. this is the first report of its kind in a llama.201021197211
llama heavy-chain antibody fragments efficiently remove toxic shock syndrome toxin 1 from plasma in vitro but not in experimental porcine septic shock.staphylococcus aureus produces the superantigen toxic shock syndrome toxin 1 (tsst-1). when the bacterium invades the human circulation, this toxin can induce life-threatening gram-positive sepsis. current sepsis treatment does not remove bacterial toxins. variable domains of llama heavy-chain antibodies (vhh) against toxic shock syndrome toxin 1 ([alpha]-tsst-1 vhh) were previously found to be effective in vitro. we hypothesized that removing tsst-1 with [alpha]-tsst-1 vhh hemofiltration filter ...201020016410
ovarian follicular dynamics and hormonal secretory profiles in guanacos (lama guanicoe).the objective of the present study was to describe ovarian activity in 11 adult non-mated guanacos, evaluated every second day for 40 days by transrectal ultrasonography and by plasma estradiol-17beta and progesterone concentrations. an inverse relationship (r = -0.29, p < 0.001) was observed between the diameter of the largest ovarian follicle and the total number of follicles indicating that follicular growth in guanacos occurs in waves. the mean duration of follicular wave was 15.1 +/- 4.2 da ...201020022436
single domain antibodies are specially suited for quantitative determination of gliadins under denaturing conditions.food intended for celiac patients' consumption must be analyzed for the presence of toxic prolamins using high detectability tests. though 60% ethanol is the most commonly used solvent for prolamins extraction, 2-mercaptoethanol (2-me) and guanidinium chloride (guhcl) can be added to increase protein recovery. however, ethanol and denaturing agents interfere with antigen recognition when conventional antibodies are used. in the present work, a new method for gliadins quantification is shown. the ...201020039674
antibody-mediated purification of co-expressed antigen-antibody complexes.immunoaffinity is an established chromatographic method for isolating macromolecules independently on the presence of specific tags while the tight interaction between antigen and antibody has been exploited to stabilize proteins during crystallization trials. therefore, it seems reasonable to try to combine the two protocols, namely to co-express the target proteins together with their specific antibodies to obtain stable complexes suitable for direct purification and further analyses. using th ...201020060475
llama single domain antibodies specific for the 7 botulinum neurotoxin serotypes as heptaplex immunoreagents.there are currently 7 known serotypes of botulinum neurotoxin (bont) classified upon non-cross reactivity of neutralizing immunoglobulins. non-neutralizing immunoglobulins, however, can exhibit cross-reactivities between 2 or more serotypes, particularly mosaic forms, which can hamper the development of highly specific immunoassays, especially if based on polyclonal antisera. here we employ facile recombinant antibody technology to subtractively select ligands to each of the 7 bont serotypes, re ...201020098614
single domain antibody templated nanoparticle resistors for sensing.the diversity of biomolecules enables materials to be synthesized at ambient conditions into complex shapes and structures while affording improved control over nanoparticle synthesis, structures, properties, and functionality. also, biomolecules can impart recognition and sensing capabilities to metal nanoparticles. here, llama single domain antibodies selected towards the beta-subunit of cholera toxin were used to template the synthesis of gold nanoparticles and subsequently assembled onto ele ...201020117926
a single-domain llama antibody potently inhibits the enzymatic activity of botulinum neurotoxin by binding to the non-catalytic alpha-exosite binding region.ingestion or inhalation of botulinum neurotoxin (bont) results in botulism, a severe and frequently fatal disease. current treatments rely on antitoxins, which, while effective, cannot reverse symptoms once bont has entered the neuron. for treatments that can reverse intoxication, interest has focused on developing inhibitors of the enzymatic bont light chain (bont lc). such inhibitors typically mimic substrate and bind in or around the substrate cleavage pocket. to explore the full range of bin ...201020138889
a llama-derived gelsolin single-domain antibody blocks gelsolin-g-actin interaction.rna interference has tremendously advanced our understanding of gene function but recent reports have exposed undesirable side-effects. recombinant camelid single-domain antibodies (vhhs) provide an attractive means for studying protein function without affecting gene expression. we raised vhhs against gelsolin (gsnvhhs), a multifunctional actin-binding protein that controls cellular actin organization and migration. gsnvhh-induced delocalization of gelsolin to mitochondria or the nucleus in mam ...201020140750
coccidioides posadasii keratouveitis in a llama (lama glama).a 7-year-old male llama was examined for a 3-month history of weight loss, and unilateral keratouveitis. clinical examination revealed nonulcerative corneal stromal abscessation, corneal vascularization, corneal edema, miosis, posterior synechia, cataract, and fibrin in the anterior chamber of the right eye. the left eye was normal. histopathology of the right eye following enucleation revealed pyogranulomatous keratouveitis with intralesional fungal spherules consistent with coccidioides spp. p ...201020149177
oestradiol-17beta plasma concentrations after intramuscular injection of oestradiol benzoate or oestradiol cypionate in llamas (lama glama).llamas (lama glama) are induced ovulators and the process of ovulation depends on dominant follicular size. in addition, a close relationship between behavioural estrus and ovulation is not registered in llamas. therefore, the exogenous control of follicular development with hormones aims to predict the optimal time to mate. oestradiol-17beta (e2) and its esters are currently used in domestic species, including camelids, in synchronization treatments. but, in llamas, there is no reports regardin ...201020149242
effect of estradiol benzoate used at the start of a progestagen treatment on superovulatory response and embryo yield in lactating and non-lactating llamas.two experiments were conducted to determine the effect of estradiol benzoate (eb) and intravaginal progestagen treatment on ovarian follicular dynamics and superstimulatory response in ecg-treated llamas. the purpose of experiment 1 was to evaluate the effect of eb and progestagen treatment starting at different phases of dominant follicle (df1) development on regression pattern and subsequent follicle wave emergence (we2) in lactating and non-lactating llamas. early lactating (n=24, 30+/-4 days ...201020189735
structural basis for a pabpn1 aggregation-preventing antibody fragment in opmd.oculopharyngeal muscular dystrophy is caused by small alanine expansions in polyadenylate binding protein nuclear 1 (pabpn1) protein resulting in its intranuclear accumulation in skeletal muscle. 3f5 llama antibody specifically interferes with the pabpn1 aggregation process in vitro and in vivo. to understand the structural basis for its epitope recognition we mapped the binding interface of 3f5 with pabpn1 and provide a structural model of the 3f5-pabpn1 complex. we show that 3f5 complementarit ...201020226184
a candidate gene for choanal atresia in alpaca.choanal atresia (ca) is a common nasal craniofacial malformation in new world domestic camelids (alpaca and llama). ca results from abnormal development of the nasal passages and is especially debilitating to newborn crias. ca in camelids shares many of the clinical manifestations of a similar condition in humans (charge syndrome). herein we report on the regulatory gene chd7 of alpaca, whose homologue in humans is most frequently associated with charge. sequence of the chd7 coding region was ob ...201020237599
estimating total body water content in suckling and lactating llamas (lama glama) by isotope dilution.total body water (tbw) in 17 suckling and six lactating llamas was estimated from isotope dilution at three different post natum and lactation stages using both (18)o and deuterium oxide (d(2)o). in total, 69 tbw measurements were undertaken. while tbw in lactating dams, expressed in kilogram, remained stable during the three measurement periods (91.8 +/- 15.0 kg), the body water fraction (tbw expressed in percent of body mass) increased slightly (p = 0.042) from 62.9% to 65.8%. in contrast, tbw ...201020379776
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