Publications

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 ...	2010	20940265
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.	2010	20959824
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 ...	2010	20969962
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 ...	2010	21088185
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.	2010	21155679
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.	2010	21175689
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.	2010	21197211
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 ...	2009	20468081
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.	2009	19650704
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 ...	2009	19616837
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 ...	2009	19557816
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 ...	2009	19562286
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 ...	2009	19597828
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 ...	2009	19082566
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 ...	2009	19085971
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 ...	2009	19121542
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 ...	2009	19132876
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 ...	2009	19171923
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.	2009	19178480
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 ...	2009	19183352
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 ...	2009	19196718
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.	2009	19228355
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 ...	2009	19258344
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 ...	2009	19269688
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 ...	2009	19286507
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 ...	2009	19286516
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 ...	2009	19349120
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 ...	2009	19356144
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 ...	2009	19359042
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 ...	2009	19368770
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 ...	2009	19392664
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), ...	2009	19446959
alpaca and llama health management. preface.		2009	19460637
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 ...	2009	19460645
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.	2009	19460650
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 ...	2009	19475676
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 ...	2009	19523913
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 ...	2009	19531051
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 ...	2009	18677581
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.	2009	19801806
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. ...	2009	19819627
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 ...	2009	19744487
use of serological techniques for diagnosis of mycobacterium bovis infection in a llama herd.		2009	19749210
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 ...	2009	19717575
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 ...	2009	19686537
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 ...	2009	20015576
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 ...	2009	19824883
[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 ...	2009	19831309
a review of the nutritional content and technological parameters of indigenous sources of meat in south america.	meat yields, proximate compositions, fatty acids compositions and technological parameters are reviewed for species which might be further developed as indigenous sources of meat in south america. these include the alpaca (lama pacos), capybara (hydrochoerus hydrochaeris), guanaco (lama guanicoe), llama (lama glama), nutria (myocastor coypus), collared peccary (tayassu tajacu), greater rhea (rhea americana), lesser rhea (rhea pennata), yacare (caiman crocodilus yacare), tegu lizard (tupinambis m ...	2008	22063568
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 ...	2008	18692907
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 ...	2008	18697767
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 ...	2008	18755196
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 ...	2008	18791271
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 ...	2008	18803786
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 ...	2008	18842738
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 ...	2008	18855250
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 ...	2008	18947189
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 ...	2008	18949046
placental glycosylation in a cama (camel-llama cross) and its relevance to successful hybridisation.		2008	18950721
parasite-induced leucomyelopathy in llamas (lama glama).		2008	18953077
probing of actinobacillus pleuropneumoniae apxiiia toxin-dependent cytotoxicity towards mammalian peripheral blood mononucleated cells.	abstract:	2008	19046441
the effect of temperature variation on biomethanation at high altitude.	the aim of the current study was to examine effects of daily temperature variations on the performance of anaerobic digestion. forced square-wave temperature variations (between 11 and 25, 15 and 28, and 19 and 32 degrees c) were imposed on a bench-scale digester using a mixture of llama-cow-sheep manure in a semi-continuous process. the volumetric biogas production rate, methane yield, and the volatile solid reductions were compared with the results obtained from anaerobic digestion (ad) at con ...	2008	18262414
spect imaging with 99mtc-labeled egfr-specific nanobody for in vivo monitoring of egfr expression.	overexpression of the epidermal growth factor receptor (egfr) occurs with high incidence in various carcinomas. the oncogenic expression of the receptor has been exploited for immunoglobulin-based diagnostics and therapeutics. we describe the use of a llama single-domain antibody fragment, termed nanobody, for the in vivo radioimmunodetection of egfr overexpressing tumors using single photon emission computed tomography (spect) in mice.	2008	18297364
corneal epithelial inclusion cyst in a llama.	a 13-year-old, female llama presented for evaluation of a limbal based corneal mass involving the od of 4 months duration. the mass was excised en bloc by a nonpenetrating keratectomy, followed by placement of a conjunctival advancement flap covering the keratectomy site. the mass was submitted for histological evaluation. histopathology identified the mass to be a corneal epithelial inclusion cyst filled with necrotic squamous and neutrophilic debris. surgical excision was complete and consider ...	2008	18302575
cerebral and renal candidosis in a llama (lama glama).		2008	18408199
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 ...	2008	18460625
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 ...	2008	18495344
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 ...	2008	18534789
alpaca and llama nematodes in britain.		2008	18567938
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.	2008	18624072
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 ...	2008	18628305
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 ...	2008	18632156
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 ...	2008	18632867
alpaca and llama nematodes in britain.		2008	18660528
collection method, season and individual variation on seminal characteristics in the llama (lama glama).	the objective of the present study was to evaluate the effect of semen collection method (electroejaculation "ee" as compared with the artificial vagina "av"), the season (summer versus winter) and the male used on macroscopic and microscopic characteristics of ejaculates in llamas. a total of 110 ejaculates were collected from six males and 92 of them were analyzed. ejaculate volume, concentration, total sperm and the following sperm characteristics were studied: motility, membrane function (ho ...	2008	17383121
carbon monoxide: a novel pulmonary artery vasodilator in neonatal llamas of the andean altiplano.	to study the nitric oxide (no) and carbon monoxide roles in the regulation of the pulmonary circulation in lowland and highland newborn sheep and llamas.	2008	18006479
in vitro production of llama (lama glama) embryos by ivf and icsi with fresh semen.	the interest for south american camelids has increased in the last years. the aim of the present research was to compare the in vitro production of lama glama embryos using two techniques: in vitro fertilization (ivf) and intracytoplasmic sperm injection (icsi). for ivf technique, we compared the effect of adding or not, heparin, penicillamine and hypotaurine as sperm capacitating agents. in the oocyte group subjected to icsi, activation with or without, ionomycin and 6-dimethylaminopurine (6-dm ...	2008	18054452
isolation and characterization of anti-fcgammariii (cd16) llama single-domain antibodies that activate natural killer cells.	fcgammariii (cd16) plays an important role in the anti-tumor effects of therapeutic antibodies. bi-specific antibodies (bsabs) targeting fcgammariii represent a powerful alternative to the recruitment of the receptor via the fc fragment, but are not efficiently produced. single-domain antibodies (sdabs) endowed with many valuable structural features might help to bypass this problem. in the present work, we have isolated anti-fcgammariii sdabs (c21 and c28) from a phage library generated from a ...	2008	18073223
a versatile nanotrap for biochemical and functional studies with fluorescent fusion proteins.	green fluorescent proteins (gfps) and variants thereof are widely used to study protein localization and dynamics. we engineered a specific binder for fluorescent proteins based on a 13-kda gfp binding fragment derived from a llama single chain antibody. this gfp-binding protein (gbp) can easily be produced in bacteria and coupled to a monovalent matrix. the gbp allows a fast and efficient (one-step) isolation of gfp fusion proteins and their interacting factors for biochemical analyses includin ...	2008	17951627
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). ...	2008	18155895
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.	2008	19183735
multiplexed fluid array screening of phage displayed anti-ricin single domain antibodies for rapid assessment of specificity.	phage display is a well-known technique that facilitates the selection of peptides or proteins that bind to a desired target. using this tool, binding elements contained in the natural immune repertoires of the source animal or from a synthetically generated collection may be selected. the unpaired variable domain of the llama's heavy-chain-only classes of immunoglobulins represents an ideal source of genetic material to create phage display libraries. initial panning of a semi-synthetic llama l ...	2007	18251258
human and animal infections with mycobacterium microti, scotland.	during 1994-2005, we isolated mycobacterium microti from 5 animals and 4 humans. only 1 person was immunocompromised. spoligotyping showed 3 patterns: vole type, llama type, and a new variant llama type.	2007	18258049
a fungal granuloma of the frontal sinus in a llama.	a 12-year-old, castrated male llama (lama glama) presented with a 12-cm diameter cranial mass. computed tomography and postmortem examination revealed that the mass invaded the calvarium and compressed the rostral part of the brain. light microscopic examination confirmed a fungal granuloma.	2007	17966336
[congenital malformation of the front limbs in a llama (lama glama): polydactyly in combination with arthrogryposis].	a male, 10 month old llama with malformation of both front limbs was presented. both front limbs had one more digit located medially. the distal phalanx of this additional digit at the left front limb reached the ground by the tip of the keratinzed pad and the toenail. the accessory digit at the right front limb was bent in a 90 degree angle caudolateraly. beside the digital bones of the accessory digits the second metacarpal bone and the first carpal bone could be detected in both front limbs b ...	2007	18085162
surgical repair of femoral fractures in new world camelids: five cases (1996-2003).	five new world camelids were admitted to the western college of veterinary medicine between 1996 and 2003 for evaluation of femoral fractures. there were three alpacas and two llamas. four of the animals were female and three were less than 3 months of age. fracture configurations consisted of distal physeal fractures (three), a comminuted diaphyseal/metaphyseal fracture, and a transverse diaphyseal fracture. fractures were diagnosed with a combination of physical examination and radiographs in ...	2007	17397385
prevalence of neoplasia in llamas and alpacas (oregon state university, 2001-2006).	prevalence and type of neoplastic disease were determined in 551 camelid submissions (368 alpacas [lama pacos], 180 llamas [lama glama], and 3 cases in which species was not identified) over a 5-year period. forty neoplasms were identified in 38 animals (6.9%). prevalence of neoplasia in llamas was higher (11%) than in alpacas (4.9%). mean age of camelids with neoplasia was 9.42 +/- 4.9 years. mean age of alpacas with neoplasia (5.48 +/- 3.7 years) was significantly less than of llamas with neop ...	2007	17402619
lactobacillli expressing llama vhh fragments neutralise lactococcus phages.	bacteriophages infecting lactic acid bacteria (lab) are widely acknowledged as the main cause of milk fermentation failures. in this study, we describe the surface-expression as well as the secretion of two functional llama heavy-chain antibody fragments, one binding to the major capsid protein (mcp) and the other to the receptor-binding proteins (rbp) of the lactococcal bacteriophage p2, by lactobacilli in order to neutralise lactococcal phages.	2007	17875214
selection of genetically encoded fluorescent single domain antibodies engineered for efficient expression in escherichia coli.	genetically encoded fluorescent antibodies are desirable for many applications in biotechnology and proteomics. however, the efficient production of single chain antibodies fused to fluorescent proteins like green fluorescent protein is still a major challenge. this is due to the opposite redox requirements of recombinant antibodies and fluorescent proteins for proper folding. to overcome this obstacle, we designed a novel strategy to directly select functional fluorescent antibodies (fluobodies ...	2007	17921141
evaluation of neospora caninum and toxoplasma gondii infections in alpaca (vicugna pacos) and llama (lama glama) aborted foetuses from peru.	the aim of this study was to investigate the participation of neospora caninum and toxoplasma gondii in abortion cases of peruvian llamas and alpacas. fifteen aborted foetuses were recovered from two main rearing areas of camelids in peru (central or south andean region). foetal histopathology was used to detect the presence of protozoal-associated lesions in target organs. n. caninum and t. gondii infections were confirmed by immunohistochemistry (ihc) combined with pcr and by pcr alone, respec ...	2007	17931783
stabilization of an immunoglobulin fold domain by an engineered disulfide bond at the buried hydrophobic region.	we report for the first time the stabilization of an immunoglobulin fold domain by an engineered disulfide bond. in the llama single-domain antibody, which has human chorionic gonadotropin as its specific antigen, ala49 and ile70 are buried in the structure. a mutant with an artificial disulfide bond at this position showed a 10 degrees c higher midpoint temperature of thermal unfolding than that without the extra disulfide bond. the modified domains exhibited an antigen binding affinity compara ...	2007	17932041
measurements of the bodyweight and other physical characteristics of 11 llamas (lama glama) from birth to weaning.	the bodyweight (bw), thoracic circumference, length and height of 11 llamas were recorded weekly from birth for 27 weeks. their mean (sd) birth weight was 11.60 (2.35) kg and their weight increased in curvilinear fashion reaching 61.05 (13.75) kg after 27 weeks. their average daily gains reached the highest value three weeks after birth and then decreased, reaching the lowest value of 262 g per day at 27 weeks. on a metabolic bw(0.83) basis, the growth of the llamas when their dams were at peak ...	2007	17938410
adaptations of subpalpebral lavage systems used for llamas (lama glama) and a harbor seal (phoca vitulina).	subpalpebral lavage systems (splss) were adapted for use in zoo llamas (lama glama) and a wild harbor seal (phoca vitulina) during therapy for severe ulcerative keratitis or corneal perforation. one llama presented with a melting corneal ulcer caused by pseudomonas aeruginosa, which necessitated frequent application of a topical ophthalmic antibiotic. the lavage system was used routinely during the day and was connected to a balloon infusion system at night to allow for continuous medication adm ...	2007	17939355
rapid assembly of sensitive antigen-capture assays for marburg virus, using in vitro selection of llama single-domain antibodies, at biosafety level 4.	there is a pressing need for rapid and reliable approaches to the delivery of sensitive yet rugged diagnostic assays specific for emerging viruses, to hasten containment of outbreaks when and wherever they occur. within 3 weeks, we delivered an antigen-capture assay for marburg virus (marv) that was based on llama single-domain antibodies (sdabs) selected at biosafety level 4. four unique sdabs were capable of independently detecting marv variants musoke, ravn, and angola without cross-reactivit ...	2007	17940952
in vitro production of llama (lama glama) embryos by intracytoplasmic sperm injection: effect of chemical activation treatments and culture conditions.	assisted reproductive technologies in the llama (lama glama) are needed to provide alternative methods for the propagation, selection and genetic improvement; however, recovery of adequate quantity and quality of spermatozoa for conventional ivf is problematic. therefore, an effort was made to adapt the intracytoplasmic sperm injection (icsi) procedure for the in vitro production of llama embryos. the specific objectives of this study were: (1) to determine in vitro maturation rates of oocytes r ...	2007	16846701
generation of llama single-domain antibodies against methotrexate, a prototypical hapten.	single-domain antibodies specific to methotrexate (mtx) were obtained after immunization of one llama (llama glama). specific vhh domains (v-d-j-region) were selected by panning from an immune-llama library using phage display technology. the antibody fragments specific to mtx were purified from escherichia coli (c41 strain) periplasm by immobilized metal affinity chromatography with an expression level of around 10mg/l. a single band around 16,000da corresponding to vhh fragments was found afte ...	2007	17011035
estradiol production by preimplantation blastocysts and increased serum progesterone following estradiol treatment in llamas.	estradiol is a potential candidate for the blastocyst signal responsible for maternal recognition of pregnancy in the llama (lama glama). two experiments were conducted to determine if the llama blastocyst produces estradiol during the presumed period of maternal recognition of pregnancy and if exogenous estradiol can extend the luteal phase. in experiment 1, llamas were superovulated with ecg and mated 7 days later (day 0=day of mating). blastocysts were collected nonsurgically on days 7, 9, or ...	2007	17116376
passive immunization of guinea pigs with llama single-domain antibody fragments against foot-and-mouth disease.	foot-and-mouth disease (fmd) is a highly contagious disease that occasionally causes outbreaks in europe. there is a need for therapies that provide rapid protection against fmd in outbreak situations. we aim to provide such rapid protection by passive immunization with llama single-domain antibody fragments (vhhs). twenty-four vhhs binding serotype o fmdv in vitro were isolated from immunized llamas by phage display and expressed in bakers yeast for further characterization. they recognized fou ...	2007	17127019
a novel pentamer versus pentamer approach to generating neutralizers of verotoxin 1.	verotoxins (vts), or shiga-like toxins, are produced by enterohemorrhagic escherichia coli (ehec), which cause hemorrhagic colitis and hemolytic uremic syndrome. vts are the major virulence factors in ehec infection due to their cytotoxicity to various types of cells. here, we present a novel type of vt neutralizer based on pentavalent single-domain antibodies, or pentabodies. two single-domain antibodies (sdabs) specific for the receptor binding sites of the b subunit of vt1 (vt1b) were isolate ...	2007	17134756
humerus fractures in llamas and alpacas: seven cases (1998-2004).	to describe treatment and outcome of humerus fractures in llamas and alpacas.	2007	17214823